Breeding and Genetics

#### **Chapter 6**

## The Focus on Core Genetic Factors That Regulate Hepatic Injury in Cattle Seems to Be Important for the Dairy Sector's Long-Term Development

*Avishek Mandal*

#### **Abstract**

The cattle during the perinatal period, as well as malnutrition, generate oxidative stress which leads to high culling rates of calves after calving across the world. Although metabolic diseases have such a negative impact on the welfare and economic value of dairy cattle, that becomes a serious industrial concern across the world. According to research, genetic factors have a role or controlling fat deposition in the liver by influencing the biological processes of hepatic lipid metabolism, insulin resistance, gluconeogenesis, oxidative stress, endoplasmic reticulum stress, and inflammation, all of which contribute to hepatic damage. This review focuses on the critical regulatory mechanisms of VEGF, mTOR/AKT/p53, TNF-alpha, Nf-kb, interleukin, and antioxidants that regulate lipid peroxidation in the liver via direct or indirect pathways, suggesting that they could be a potential critical therapeutic target for hepatic disease.

**Keywords:** cattle liver, inflammation of the liver, gene expression, antioxidant, cytokine

#### **1. Introduction**

The "oxygen paradox," which happens when free radicals (RL) are produced during mitochondrial respiration, is supported by a huge body of research that shows that, despite the necessity of oxygen for life, it also has a damaging effect on the body [1]. Now it is interesting to learn what changes oxidation conditions in cattle or bovine liver. Since oxidative stress is the root cause of several illnesses in cattle, such as sepsis, mastitis, enteritis, pneumonia, and respiratory and joint issues, its effects on food are well-known, but they are also, gradually, recognized to have impacts on the organism "in vivo" [2]. Numerous studies have demonstrated the significance of providing antioxidants for animal nutrition and their connection to oxidative stress, taking into consideration the significance at each stage [3, 4]. Consequently, we can state that the

antioxidant impact will not only improve the health of the animals but also raise the quality of the finished product (meat and milk). The large amount of non-esterified fatty acids absorbed by the liver exceeds its ability for oxidation and, as a result, encourages liver-related illnesses including fatty liver and ketosis. Additionally, early breastfeeding cows' livers experience metabolic stress due to high rates of hepatic gluconeogenesis, which produces glucose for lactose in the mammary gland [5, 6]. In addition to this metabolic stress, early-lactating cows experience a variety of inflammatory challenges, including microbial components (lipopolysaccharides, or LPS), pro-inflammatory cytokines (tumor necrosis factor a (TNF-α), interleukin (IL)-1b, and IL-6), and reactive oxygen species (ROS), as a result of infectious diseases like mastitis and endometritis, as well as subacute [7]. As a result, transition dairy cows experience a state similar to inflammation, which is demonstrated by the production of an acute phase response (APR).The production of positive acute phase proteins (APPs), such as serum amyloid A (SAA), haptoglobin (HP), or C-reactive protein (CRP), which compete with the production of negative APPs, or essential liver proteins, such as albumins, enzymes, lipoproteins, transferring or carriers of vitamins (such as retinol-binding protein), and hormones, is a hallmark of the APR [8]. Thus, the creation of an inflammatory process in the liver exacerbates the biologically existing metabolic load in the liver of early nursing dairy cows, which impairs liver function. Decreased milk production and lower reproductive efficiency in dairy cows are both linked to low blood levels of negative APPs, which are a sign of a severe inflammatory reaction in the liver. Additionally, it has been noted that low-level intravenous TNF-alpha injection causes triacylglycerol build-up and hepatic inflammation in dairy cows [9]. In early lactation, cows with a low "liver functioning index" (LFI) have a significant inflammatory response that is characterized by an obvious increase in positive APPs, a notable drop in negative APPs, poor immunological function, and increased clinical issues [10]. Alternatively, cows with a high LFI display a reduced inflammatory response, better liver function, a slower increase in positive APPs, a slower decline in negative APPs, and fewer clinical issues during this phase [11]. This suggests that these cows have a greater ability to handle the inflammatory challenges that arise during the periparturient phase and are less likely to develop liver-related diseases. There is a lack of knowledge on the molecular underpinnings of liver-related disorders and the variations in susceptibility that exist across individuals. A better understanding of the molecular mechanisms underlying liver-associated diseases in transition dairy cows, however, may help to develop ways to avoid the occurrence of liver-associated disorders and increase production in dairy cows given that the occurrence of liver-associated diseases in dairy cows is critical because it impairs the metabolic function of the liver, overall health status, and productive and reproductive performance.

#### **2. Pathogenesis liver disease in cattle**

The build-up of excessive levels of free fatty acids (FFA) in blood or triglycerides (TAG) deposited in the liver are the main contributors to the pathogenesis of fatty liver in newborn dairy cows. In animals, the liver, which is an essential organ, controls the metabolic balance of protein, fat, and carbohydrates. Dairy cows' food consumption continues to decline after birthing, but their lactation gradually increases. As a result, the cow's body might quickly experience an inadequate supply of sugar due to the absorption of lactose, which encourages the liver to mobilize fat. The liver serves

#### *The Focus on Core Genetic Factors That Regulate Hepatic Injury in Cattle Seems to Be… DOI: http://dx.doi.org/10.5772/intechopen.108151*

as the primary location for the metabolism of both substances and energy [12]. A negative nutritional balance is improved by the rising fat mobilization, which also encourages gluconeogenesis, raises blood sugar levels, and increases blood sugar concentration. The liver's production of non-esterified fatty acids (NEFA), which are partially re-esterified to create triglycerides (TAG), a kind of very-low-density lipoprotein (VLDL) that is seldom ever transported beyond the liver, increases dramatically as a result of the increased fat mobilization. Because dairy cattle lack esterase, TAG accumulates abundantly, making them more susceptible to fatty liver disease [13, 14]. Non-alcoholic fatty liver disease (NAFLD) in people is characterized by aberrant lipid build-up in the liver, raised fasting aminotransferase (AST/ALT), and/or triglycerides (TAG) levels, increased plasma insulin and fatty acid concentration, and metabolic disorder syndromes [15]. Histological evidence of hepatic inflammation brought on by acute inflammation and subacute inflammation is also one of the most significant risk factors. Dairy cows with fatty liver disease are a classic animal model for NAFLD, useful for illuminating its pathology and etiology [16]. A "two-hit" idea has been put out by researchers recently to explain the pathogenic processes of NAFLD. (1) Insulin resistance was the "first hit's" cause (IR). In addition to causing hyperinsulinemia, IR can intensify the lipolysis of nearby tissues. Adipose tissue lipolysis causes the liver's production of TAG and FFA to rise [17, 18]. The FFA is harmful to hepatocellular function, increasing cell membrane permeability and impairing mitochondrial activity by inhibiting associated enzymes [19]. The imbalance between the coexisting oxidation and anti-oxidation processes in the liver was what led to the "second hit." Reactive oxygen species (ROS) are produced persistently as a result of increased lipid peroxidation. Other new or additional variables, such as inflammatory cytokines, adipokines, endotoxins, and mitochondrial inactivation, might boost lipid peroxidation for a second blow to the liver in addition to the preexisting components associated with the increased oxygen stress. The second hit will eventually result in the advancement of NASH (non-alcoholic steatohepatitis), which promotes oxidative stress, inflammation, cell death, and fibrosis beyond hepatic steatosis [20]. Particularly, the inflammation inhibits lipase activity, prevents the transit of lipids and/or lipoproteins, and results in lipid build-up, which is negatively connected with lipolysis and positively correlated with liver damage [21]. Alternatively, it causes cell apoptosis, IR, and lipid peroxidation, exacerbating the pathophysiology of NAFLD. (3) Hepatocyte cell death and irreversible cell repair constitute a "third hit," in fact. (4) In addition, endoplasmic reticulum (ER) stress is another significant "hit" in the pathophysiology of NAFLD. Obesity and diabetes, two metabolic illnesses, can result in ER stress, which impairs the physiological processes of liver cells by causing an accumulation of improperly folded proteins (unfolded protein response, UPR) [22]. It is important to note that ER stress can cause SREBP (sterol-regulatory element binding protein) to become active. This promotes the transcription of acetyl-CoA carboxylase (ACC) and fatty acid synthase (FAS), which leads to an increase in the production of TAG and fatty acids in the liver [23]. Furthermore, the production of ROS by liver cells under oxidative stress can cause ER stress, which can result in improper protein folding and/or protein modification. Oxidative stress can also be brought on by ER stress. Through many mechanisms, the biological processes of ER stress and oxidative stress interact with one another, causing IR and exacerbating NAFLD. The pathophysiology of NAFLD is largely unknown, though [24, 25]. It was believed that the key route and/or the primary risk factors implicated in the etiology of NAFLD are abnormalities in lipid and lipoprotein metabolism coupled with chronic inflammation and oxidative stress. The prevalence of fatty liver is widely thought to be linked

to biological processes such as disordered glycometabolism, oxidative stress, and intracellular inflammatory response in addition to being directly tied to insulin resistance and fat metabolism issue. Additionally, these processes are connected to and/or coordinated with one another, which accelerates the development of NAFLD [26, 27].

NAFLD's pathophysiology is largely unknown, though. It was believed that the key route and/or the primary risk factors implicated in the etiology of NAFLD are abnormalities in lipid and lipoprotein metabolism coupled with chronic inflammation and oxidative stress. Additionally, these processes are connected to and/or coordinated with one another, which quickens the development of NAFLD. The pathophysiology of NAFLD is influenced by a wide range of variables. In the hepatic lipid metabolism, lipid transport, and secretion, for instance, PPAR (peroxisome proliferator-activated receptor) and/or PPAR, microsomal triglyceride transport protein (MTP), and apolipoprotein (apo B) play crucial roles [28–30]. Tumor necrosis factor, leptin, and adiponectin are examples of adipocytokines. Cytokines include interleukin-6 (IL-6), glucagon-like peptide-1 (GLP-1), fibroblast growth factor 19 (FGF-19), and fibroblast growth factor 21 (FGF-21), growth hormone-releasing hormone (GHRH), and others [31–33]. Toll-like receptors (TLRs) also play a role in insulin resistance. It has been suggested that microRNAs (such as mir-107 and miR-103) control insulin resistance [34–36]. The antagonist against microRNA-103/107, RG-125 (also known as AZD4076), just started a phase I clinical trial to treat NASH (non-alcoholic steatohepatitis) [37]. As a result, aberrant hepatic lipid metabolism, gluconeogenesis, oxidative stress, and inflammation are frequently linked to the etiology of NAFLD. These biological processes' causal connections and underlying molecular mechanisms are yet unknown. However, it would be beneficial to understand the molecular etiology of NAFLD if some significant regulatory factors or genes that control all these biological processes were discovered.

#### **3. Vascular endothelial growth factor (VEGF)**

Native VEGF is a homodimeric glycoprotein of 45 kDa that may bind to heparin and stimulate the proliferation of vascular endothelial cells generated from arteries, veins, and lymphatics [38, 39]. It is expressed by a single gene. Even though VEGF primarily targets endothelial cells, studies have shown that several non-endothelial cell types, such as retinal pigment epithelial cells, pancreatic duct cells, Schwann cells, and maybe placental cell functions, are also subject to mitogenic effects. Five distinct VEGF isoforms, with respective amino acid compositions of 120, 145, 165, 188, and 205, are produced [40] as a result of alternative exon splicing. VEGFR-1/ Flt-1, or Fms-like tyrosine kinase 1, and VEGFR-2/KDR, or kinase insert domaincontaining region, are the two tyrosine kinase receptors that mediate the biological functions of VEGF. Both VEGFR-1 and VEGFR-2 have a single transmembrane region, seven Ig-like structures in the extracellular domain, and a tyrosine kinase sequence that is broken by a kinase-insert domain [41, 42]. Another RTK in the same family, VEGFR-3 or Flt-4, has affinities for VEGF-C and VEGF-D. In addition to RTKs, VEGF also interacts with the neuropilin family of coreceptors [43]. The two different domains of the VEGF molecule, which are found on the opposing terminal of the VEGF monomer, allow VEGF to interact with its receptors, Flt-1 and KDR. While the 120, 145, 165, and 188 isoforms of VEGF activate KDR, VEGF165 and VEGF188 activate Flt-1 [40]. The VEGF system was found in the uterine epithelium, trophoblast, vascular tissue, and uterine glands of placentomes [44]. Bovine VEGF

#### *The Focus on Core Genetic Factors That Regulate Hepatic Injury in Cattle Seems to Be… DOI: http://dx.doi.org/10.5772/intechopen.108151*

may have the following traditional roles in angiogenesis and vascular permeability, as well as chemotactic activity in endothelial capillaries, autocrine influence on the migration of giant trophoblastic cells, which promotes maternal-fetal interchange, and modulatory action in trophoblastic function, specifically in steroidogenesis [45–47]. These functions are all suggested by the presence of the VEGF system in the maternal-fetal interface and the vascular system. The improvement of reproductive function and productive efficiency is crucial while growing dairy and meat cattle. Numerous studies have shown that cows, especially those with high milk output, gradually exhibit a rise in reproductive issues, which are reportedly caused by several factors, including poor energy balance and unequal gene expression. In addition to facilitating the transmission of physiological information between the mother and the fetus, the placenta produces and secretes steroid hormones, such as progesterone and estrogens, which are in charge of optimizing the environment for fetal growth. Therefore, the success of gestation depends on its efficient operation. The location of these growth factors and their corresponding receptors in non-endothelial cells, however, suggests that they are involved in other physiological processes, such as the stimulation of hormone synthesis in steroidogenic tissues. NAFLD has been studied using a variety of rat models, each of which mimics one or more characteristics of human NAFLD, including steatosis, NASH, fibrosis, and HCC [48]. Numerous studies have documented how angiogenesis manifests in various models. First, it was demonstrated that mice who were given a high-fat diet (HFD) had higher levels of VEGFR-2 expression and CD31 expression, the most widely used marker of endothelial cells, in their livers [49]. In the livers of rats given a CDAA diet and mice fed an MCD diet, and cattle fed a high-fat artificial diet can cause an increase in VEGF protein was observed. Interestingly, VEGF mRNA levels remained the same in the latter scenario, pointing to posttranscriptional regulatory mechanisms [48]. The global vasculature of the cattle liver with NAFLD was examined using specific imaging methods. NAFLD was linked to an international alteration of the hepatic vascular architecture, which included not only an increase in the number of vessels but also in a visibly different phenotype of vessels, which displayed an enlarged diameter and a disrupted organization, according to research using scanned electronic microscopy of vascular corrosion casts of the liver [50].

#### **4. Practices for consuming more antioxidants**

Antioxidants may be produced by the body, obtained through food, or given orally. Some vitamins, such as vitamin K, may be produced by the ruminal and intestinal flora while vitamin D can be produced by UV radiation on the skin in ruminants [51]. Several natural feed ingredients are also high in antioxidants, such as vitamin E or precursors to vitamin A. But, due to the wide range in vitamin concentrations in feeds and exposure to sunshine, depending only on these naturally occurring quantities might put the animals in danger of deficient disorders. Additionally, many dairy farms confine their animals indoors, where they have little exposure to sunlight and fresh fodder and the majority of natural vitamins quickly deteriorate after ensilage. As a result, these animals need to be supplemented with vitamins and trace elements, albeit the needs of grazing cattle may be different from those of cattle-fed preserved forages. Additionally, it is advised to administer an additional supplementation during times of increased need, like right before calving. The addition of vitamins and minerals to the animals' diets is arguably the approach

to antioxidant supplementation that is utilized the most frequently in industrial dairy farms, particularly when premixes are added to the overall mixed ration. The injection of vitamins and trace minerals to these animals, however, facilitates the supplementation without necessitating the creation of a specific management group of cows during this period because the needs for antioxidants are increased in moments of augmented metabolisms, such as the transition period, in farms with several animals not large enough to practically implement a specific diet for similar dry cows [52]. Numerous solutions, either for individual vitamins and trace elements or for a mix of them, are offered on the market for this purpose. The two antioxidants that are most frequently found in dairy cattle diets, either separately or together, are vitamin E and selenium. As a result, the majority of study has concentrated on these compounds' effects. Chain-breaking antioxidant vitamin E, which is highly lipid-soluble, stops the spread of free radicals in plasma lipoproteins and membranes. The effects of this trace element on selenoproteins, such as GSH-Px, are attributed to its function as a cofactor [52, 53]. However, a recent study has emphasized the direct function of selenium (Se) in preventing oxidative stress (OS) and controlling immunity in dairy calves around the time of calving. At the start of lactation, cows frequently have decreased plasma levels of vitamin E [53, 54]. It is unclear why the content of vitamin E a-tocopherol drops near the end of pregnancy. A reduction in the development of various vitamin carrier proteins in the liver of dairy cows during the transition phase causes reduced plasma levels of vitamin E. This may be partly attributed to the usage of antioxidants for colostrum formation. To avoid the reduction in plasma a-tocopherol concentrations around parturition, supplementation with relatively high vitamin E levels is required [55].

#### **5. Transport stress**

Feeder's calves are more likely to acquire bovine respiratory disease (BRD) due to transportation stress, which is frequently exacerbated by the stress of weaning. The direct cost of treating BRD was \$23.60 per case, according to the USDA (2013) [56], and at some time during the feeding period, 16.2% of feedlot cattle showed indications of respiratory illness [56, 57]. Bruising and dark-cutting carcasses are two additional beef quality issues related to shipping. Cattle can become bruised when they collide with one another during transport or when they come in touch with trailer parts, especially while loading and unloading. 68.2% of the 9860 corpses that were seen at three separate slaughterhouses in the United States had bruises, and 53.5% of those injuries were along the dorsal midline, the area of the carcass with the highest economic value depends [58]. Before slaughter, prolonged stress depletes muscle glycogen reserves, resulting in a condition called dark cutting carcasses. Long-term stress before slaughter depletes muscle glycogen levels, causing a characteristic called "dark cutting carcasses." The possibility that transportation might have a detrimental impact on carcass and meat qualities has sparked interest in transportation beef quality assurance, a program that informs cattle transporters of optimum management practices [59]. A change in the prooxidant-antioxidant equilibrium favouring the former was the original definition of oxidative stress. This condensed definition was eventually expanded to mean an imbalance favouring oxidants over antioxidants, which disrupts redox signalling and causes molecular damage in addition to impairing physiological performance. Under typical biological circumstances, the antioxidant defense mechanism of the cell balances the quantity of oxidants, causing it to vary

*The Focus on Core Genetic Factors That Regulate Hepatic Injury in Cattle Seems to Be… DOI: http://dx.doi.org/10.5772/intechopen.108151*

within a specific range known as basal oxidative stress or oxidative eustress. Cell death by necrosis, apoptosis, or both may occur when oxidative stress reaches a high intensity, known as oxidative distress. When determining the level of oxidative stress in biological samples, it is crucial to take into account both sides of the equation since oxidative stress is defined in terms of the interaction between oxidants and antioxidants. Direct measurements of reactive oxygen species (ROS), oxidative changes to proteins, lipids, and nucleic acids, antioxidant enzyme activities, antioxidant concentrations, and ROS levels are a few of the indicators that are frequently evaluated.

#### **6. Antioxidant and lipid peroxidation**

Endogenous nonenzymatic biological sources and antioxidants received exogenously make up the cellular antioxidant defense system [60]. Since several antioxidants can work together to combat the oxidative offense and a lack of one specific antioxidant does not always signal that the sample's overall neutralizing ability is compromised, quantifying antioxidant components individually does not give a fair picture of the sample's antioxidant capacity [61]. As a result, several techniques have been created to determine the overall antioxidant capacity. This takes into account the overall antioxidant activity of every sample that was analyzed rather than just the total amount of detectable antioxidants [62, 63]. The following are some of the most popular analytical techniques for determining the status of all antioxidants: the overall antioxidant capacity to capture radicals comparable antioxidant power to Trolox capability for absorbing oxygen radicals the sample's ability to reduce iron or the pool of antioxidants that would be severely oxidized by a large dose of hypochlorous acid. It is not unexpected that several endogenous regulatory processes are affected by an external antioxidant supply as a result of this interaction between antioxidant compounds. Recent studies show that these processes may be responsible for some of the contentious conclusions regarding antioxidant supplementation, even if further investigation is necessary to completely understand all the regulatory pathways in dairy cattle: The transcription of genes encoding different antioxidative and cytoprotective proteins is regulated by the nuclear factor E2-related factor 2 (Nrf2), making Nrf2 necessary for the transcription of GSH-Px 2 (and likely GSH-Px 1 as well). Found that a significant increase of Nrf2 target genes with anti-oxidative capabilities occurs throughout the transition from late pregnancy to the start of breastfeeding [64, 65]. Additionally, at this time, the unfolded protein response is triggered in dairy cow livers, activating Nrf2 via the PERK pathway and enhancing the production of antioxidant enzymes and antioxidant capacity [66, 67]. These systems may be physiological defenses against tissue damage brought on by inflammation and ROS generation [68]. They, therefore, serve as crucial benchmarks for ensuring successful adaptation during the period of transition. Additionally, the fact that excessive antioxidant supplementation reduces antioxidant capacity may be explained by the endogenous regulation of antioxidant molecules, as high antioxidant doses may reduce antioxidant capacity by suppressing Nrf2 due to lower ROS levels resulting in decreased expression of antioxidant enzymes [69]. Mastitis, which frequently originates from microbial infection of the mammary gland, is the most expensive inflammatory condition in dairy cows. LPS produced from the bacterial outer membrane is the major pathogen component starting inflammatory reactions if the infection is brought on by gramnegative bacteria.

#### **7. Oxidative stress**

Saturated fatty acid excess in the mitochondria causes redox equilibrium to break down and speeds up the production of oxygen radicals. Insulin resistance and nonalcoholic FLD in non-ruminants have been linked to the lipotoxic syndrome known as oxidative stress. As discussed by many authors, the ongoing generation of ROS triggers serine/threonine kinase signalling cascades that prevent the induction of insulinstimulated insulin receptor substrate [70, 71]. In response, simple steatosis may be promoted by increased intrahepatic lipid accumulation caused by insulin resistance (i.e. non-inflammatory phenotype). The activation of Kupffer cells, which in turn activate redox-sensitive transcription factors like nuclear factor-B and upregulate pro-inflammatory TNF-α, may also be facilitated by oxidative stress. Unfortunately, NADPH oxidase and cytochrome P450 (family 2, subfamily E, polypeptide 1; commonly known as CYP2E1) are also upregulated in inflammatory steatohepatitis, which further reduces antioxidant capacity and promotes hepatocellular damage [72]. Depletion of n-3 long-chain polyunsaturated fatty acids (FA) due to poor fatty acid desaturation and increased peroxidation in the liver are additional effects of excessive ROS formation. Furthermore, polyunsaturated fatty acids (FA) that have been exposed to enzymatic (through cyclooxygenase, lipoxygenase, and CYP2E1) or non-enzymatic (by ROS) oxidation can produce oxylipins with a variety of inflammatory activities [73, 74]. For instance, proinflammatory hydroxyl-octadecadienoic acid and hydroxyl-eicosatetraenoic acid are both oxylipins produced from n-6 arachidonic acid and linoleic acid, respectively [75]. The widespread mitochondrial FA oxidation, albeit incomplete breakdown, that occurs in the transition cow with diminished antioxidant capability is probably a contributing factor in the development of ROS build-up [76]. Adipose tissue lipolysis may be boosted by oxidative stress, which will worsen the oxidant state. Unsaturated FA (fatty acid) produced from lipolysis can undergo ROS oxidation, which produces isoprostane and lipid hydroperoxide [77]. Adipose tissue releases fatty acids that are also utilized by the liver to produce hepatic -hydroxybutyrate, which may trigger p38 mitogen-activated protein kinase activity and increase hepatocyte death. Additionally, nuclear factor B may be induced by lipolytic FA in hepatocytes through ROS-dependent processes that cause inflammation [78, 79]. These results imply that FLD pathogenesis involves oxidative stress. As previously discussed, the buildup of ROS or oxylipids probably affects the immunological responses of cattle. For instance, in peripheral blood mononuclear cells from healthy transition cows, plasma oxylipin levels are associated with the production of interleukin-12 and inducible nitric oxide synthase-2 [80]. Elevations in the arachidonic acid metabolite 15-hydroxy-peroxyeicosatetraenoic acid occur in endothelial cells along with death, caspase-3 activation, leukocyte migration, and the production of inflammatory cytokines [81]. Oxylipids generated from cytochrome P450 and lipoxygenase accumulate in the plasma and adipose tissue of postpartum cows, where they may have an impact on immune cell trafficking and inflammation. Examples of such oxylipids include 5-hydroxy-eicosatetraenoic acid [82]. The significance of oxidized lipids in the development of inflammatory illnesses such as mastitis and metritis has therefore been highlighted. The significance of oxidized lipids has been studied since one of the main characteristics of inflammatory illnesses, such as mastitis and metritis, is inflammatory dysfunction [82]. For instance, hydroxyl-octadecadienoic acid accumulation and breast inflammation are two features of *Streptococcus uberis* mastitis. Oxylipid production and associated health consequences may be influenced by dietary antioxidant intake, trace mineral intake that aids in antioxidant

*The Focus on Core Genetic Factors That Regulate Hepatic Injury in Cattle Seems to Be… DOI: http://dx.doi.org/10.5772/intechopen.108151*

defense mechanisms, and the kind and quantity of FA supplied to transition cows [83]. Uncontrolled or chronic inflammatory states can be harmful, even though they often result in the recovery from infection after a controlled inflammatory phase. Therefore, after the removal of the infectious agents, a speedy resolution phase is required for a perfect acute inflammatory response. Anti-inflammatory cytokines like IL-10 and n-3 (omega-3) fatty acid derivatives like resolving and protectors are important resolving signals.

#### **8. Cytokines**

Gene activity TNF-alpha is one of the most significant cytokines involved in starting and growing the acute-phase response [84]. TNF-α is necessary for healthy liver regeneration and increases hepatic DNA synthesis by activating NF-B [85]. Numerous cell types, particularly macrophages and mast cells, generate cytokines such tumor necrosis factor (TNF), interleukin (IL)-1, and interleukin (IL)-6. By stimulating the acute phase response and activating leukocytes and endothelial cells, they play significant roles in the inflammatory response. The liver of rats that have experienced chronic starvation maintains its acute-phase reactivity. According to the new research, although the reported rise in TNFA expression between days 14 and there was small, it may have been the result of a systemic inflammatory response that was triggered within the uterus as parturition neared (i.e., higher IL-1 and IL-8 production) [86]. The cows' metabolisms were put under additional stress due to a decreased energy balance brought on by the decrease in energy intake. It is generally known that IL-1 suppresses appetite in humans, but plasma concentrations of TNF- are linked to higher energy expenditure [87, 88]. Early lactation cows were given recombinant bovine TNF- alpha, which raised blood haptoglobin, NEFA, cortisol, growth hormone, and nitric oxide while decreasing feed intake. Some of these symptoms closely reflect periparturient-period reactions that we and others have seen, as well as reactions to endotoxin treatment [89, 90]. Since proinflammatory and signaling genes are upregulated in the liver of mice that have been induced to develop fatty liver and insulin resistance by high-fat diets, TNFA upregulation in liver macrophages may act in a paracrine manner and cause potent upregulation of SAA1 in hepatocytes [91, 92]. Increased inflammation during the formation of fatty livers in transition dairy cows may be caused by NF-B-mediated proinflammatory signals [93]. Bovine recombinant When given to breastfeeding cattle, TNF- raised their blood levels of haptoglobin, NEFA, cortisol, growth hormone, and nitric oxide while decreasing their appetite for food [94]. SAA1's expression has increased by over sixfold, which is partially explained by the fact that TNF- and IL-1 boost the manufacture of positive acute phase proteins like SAA1. The acute-phase reaction causes a 1000-fold increase in SAA1 production in the liver [95]. Furthermore, it has been demonstrated in several reports that pro-inflammatory and signalling genes are upregulated in the liver of mice induced to develop fatty liver and insulin resistance by high-fat diets. This suggests that the upregulation of TNFA in liver macrophages may act in a paracrine manner and result in a potent upregulation of SAA1 in hepatocytes.

In addition to having a deleterious impact on the neurophysiological systems controlling feed intake, IL-8 and IL-1 released from the placenta directly upregulate the expression of SAA1 and TNFA in the liver [96]. Negative energy balance, hyperinsulinemia, increased adipose tissue lipolysis, and decreased feed intake all have an impact on the liver's ability to access nutrients. Blood NEFA and -hydroxybutyrate

levels may be considerably elevated and lipolysis may be further stimulated by cytokines from the liver and/or placenta (BHBA) [97]. Circulating NEFA most likely operate as endogenous ligands for PPARA and HNF4A, upregulating them and activating downstream genes involved in fatty acid oxidation, ketogenesis, and gluconeogenesis (ACSL1, ACOX1, carnitine palmitoyl transferase 1A (CPT1A), ACADVL) (PCK1) [98, 99]. The outcome of metabolic processes that were partially sparked by the overexpression of PPARA and HNF4A is net hepatic glucose synthesis and glucose and amino acids for milk synthesis [100]. Through the direct overexpression of GPAM, fatty acid synthase (FASN), ATP-citrate lyase (ACLY), and Spot 14, activation of PPARA can reduce the expression of the gene strongly connected with lipid synthesis, sterol regulatory element binding transcription factor 1 (SREBF1) (S14) [101]. Higher levels of liver triacylglycerol are correlated with the upregulation of SREBF1 via cytokines or fatty acids and GPAM [102]. Limitations in insulin and amino acid delivery to the liver may suppress IGFBP3, EIF4B, 3-phosphoinositide dependent protein kinase-1 (PDPK1), proteasome (prosome, macropain) 26S subunit, ATPase 2 (PSMC2), and/or PDPK1-dependent protein kinase, resulting in a reduction in hepatic protein synthesis, circulating blood IGF-I, and liver glycogen [103]. Increased lipid peroxidation in the liver may result from the downregulation of GSTM5 expression. The danger for hepatic periparturient health disorder is increased by both less ability to detoxify lipid peroxidation products and larger triacylglycerol build-up in the liver [104].

#### **9. Endoplasmic reticulum stress triggers cytoprotective pathways**

It's interesting to note that cryoprotective pathways, including the nuclear factor E2-related factor 2 (Nrf2) pathway, are activated by ER stress brought on by ROS or pro-inflammatory cytokines [105]. This activation is PERK-dependent. Various antioxidative and cryoprotective proteins are controlled by the redox-sensitive transcription factor Nrf2. ER stress causes Keap1 to become disassociated from Nrf2, allowing Nrf2 to move into the nucleus and activate antioxidant and cytoprotective genes by binding to antioxidant response elements in the promoter regions of its target genes. In the absence of ER stress-inducing stimuli, Nrf2 is inactive and retained in the cytoplasm by interaction with Kelch-like ECH-associated protein 1 (Keap1) [106]. The activation of Nrf2 also lowers pro-inflammatory signalling, attenuates inflammatory damage, and neutralizes ROS generated under pro-inflammatory situations, which decreases the vulnerability of tissues to oxidative damage and cytotoxicity [107]. Thus, it has been proposed that ER stress-induced activation of Nrf2 is a method for reducing oxidative damage that is brought on by ER stress.Following the presence of enhanced ROS and pro-inflammatory cytokines in the liver of transition dairy cows, we have recently noticed Nrf2 activation as demonstrated by overexpression of Nrf2 target genes, including catalase, glutathione peroxidase 3, microsomal glutathione S-transferase 3, haem oxygenase 2, metallothionein 2A, NAD(P)H dehydrogenase, quinone 1 [108]. Furthermore, it's been proposed that Nrf2 is a physiological target to preserve liver function and enhance overall health in high-yielding dairy cows. It is not unlikely that interindividual variations in the effectiveness of activating the Nrf2 pathway in response to ROS or inflammatory stimuli explain the molecular level for variations in the susceptibility to develop liver-associated diseases between earlylactating dairy cows with similar NEB and milk yield [109]. This is because Nrf2 plays a crucial role in preventing liver damage.

*The Focus on Core Genetic Factors That Regulate Hepatic Injury in Cattle Seems to Be… DOI: http://dx.doi.org/10.5772/intechopen.108151*

#### **10. Tnf-α and NF-kb**

The pattern-recognition receptor toll-like receptor 4 (TLR4), which recognizes endogenous ligands and external pathogen-associated molecular patterns, is crucial in the development of the inflammatory response [110]. The production of proinflammatory cytokines and the activation of nuclear factor kappa B (NF-B) signalling pathways in a variety of cell types are both correlated with TLR4 activation. In rat models of cardiac ischemia-reperfusion, it has been found that TLR4 expression positively correlates with the levels of tumor necrosis factor (TNF) and interleukin-6 (IL-6) [111]. In dairy cattle, disease -affected cows' milk and intra-mammary epithelial cells showed elevated NF-B activity. Lymphocyte antigen-96, also known as MD-2, and CD14 form a complex when lipopolysaccharide (LPS) binds to TLR4 [112]. This complex then triggers TIR (Toll/IL1 Receptor domain) intracellular signalling through adaptor molecules, primarily myeloid differentiation actor 88. (MyD88) [113]. This TLR4 and damage signalling causes downstream kinases to become active, which in turn causes IKB to degrade, releasing NF-B to go to the nucleus [114, 115]. In the promoter region of genes encoding pro-inflammatory cytokines, such as IL-1B and IL-6, it binds B sites [116]. Bovine mammary epithelial cells (bMEC) react differently to diverse pathogenic stressors, according to some researchers. While the reaction to *Staphylococcus aureus* culture supernatant (SaS) was linked to an AP-1 and IL-17A signalling route, crude LPS from *Escherichia coli* was linked to an NF-B and Fas signalling network [117]. The impact of intra-mammary cephapirin therapy, either alone or in combination with prednisolone, on gene expression patterns in experimentally induced mastitis in Holstein Friesian cows was examined [118]. In comparison to challenged, untreated areas, they discovered that both treatments led to a down-regulation of gene transcripts implicated in chemokine and TLR-signaling pathways. It is widely known that TLR4 is a key cell surface receptor for the inflammatory response because it recognizes LPS from the cell wall of gram-negative bacteria and starts the MyD88-IKKNF-B pathway response [119]. The MyD88-dependent pathway is activated by TLR4 regulation of LPS (Mediated by TLR-IL-1 receptor domain containing adapter protein/TIRAP), which triggers the immediate activation of NF-B and the subsequent induction of a number of pro-inflammatory cytokines [120, 121]. Additionally, it was shown that thymol may suppress NF-B activation and down-regulate the mRNA production of tracheal antimicrobial peptide and -defensin, hence reducing the internalization of *S. aureus* into bMEC (BNBD5) [122].

#### **11. AKT and mTOR**

A subset of genes involved in lysosomal biogenesis and function, as well as those involved in the creation of autophagosomes, are controlled by transcription factor EB (TFEB), a master transcription regulator [123]. Lysosome-associated membrane protein (LAMP1) and the V0 domain of the vacuolar ATPase (ATP6V0A1) are two examples of the hepatic lysosome-regulated genes that were abundantly expressed in mice after TFEB activation or overexpression. Furthermore, TFEB overexpression in HeLa cells increased the expression of autophagy-related genes including genes such sequestosome-1 (SQSTM1) and microtubule-associated protein 1 light chain 3 (MAP1LC3) (ATG12) [124]. It's important to note that studies have shown that the livers of mice and people with non-alcoholic fatty liver disease had decreased

TFEB transcriptional activity and lysosomal function [125, 126]. Furthermore, low levels of the genes MAP1LC3, SQSTM1, ATG7, and ATG12 in the liver of ketotic dairy cattle propose a reduction in the formation of autophagosomes, suggesting that impaired TFEB transcriptional activity may exist in the liver of dairy cows with ketosis and result in elevated aminotransferase enzyme levels [127]. Mechanistic target of rapamycin kinase complex 1 (mTORC1) phosphorylates the transcription factor TFEB at Ser 211 in nonruminants to prevent its subcellular localization and activity [128]. Other kinases, such as protein kinase B (Akt), glycogen synthase kinase-3 (GSK3), and extracellular signal-regulated kinase (ERK1/2), which phosphorylate TFEB at Ser 467, Ser 138, and Ser 211, respectively, also influence TFEB nuclear localization [129]. These other kinases work in conjunction with mTORC1 to affect TFEB nuclear localization. In the livers of dairy cattle in ketosis, changes in the activity of Akt, GSK3, and ERK1/2 have been observed [130, 131]. The upstream substrates of mTORC1 that are activated support anabolic pathways while blocking catabolic ones. RPS6KB, EIF4EBP1, and TFEB were more heavily phosphorylated in the current research, which is suggestive of an overactive hepatic mTORC1 state during ketosis [132]. It's probable that a similar process operates in dairy cows with ketosis as hepatic overactivation of mTORC1 lowered TFEB transcriptional activity and compromised lysosomal function in mice with fatty liver. Therefore, the decrease in molecules related to lysosomal function that researcher found in ketotic cattle may have been brought on by an overactive mTORC1 that inhibits the transcription of TFEB. It has been observed that Akt, GSK3 and ERK1/2 cause TFEB to become more phosphorylated and less likely to go into the nucleus [133]. Accordingly, in line with earlier research, phosphorylated Akt, GSK3, and ERK1/2 were reduced in the liver of dairy cows in ketosis, which decreased their impacts on TFEB transcriptional activity regulation [134].

#### **12. Conclusion**

The environment and dietary habits have a substantial influence on the health of cattle and their hepatic condition in the future. Climate change may encourage the formation of poisons or poisonous spores, which cattle are exposed to when grazing diseased grass. Using mechanical or natural methods, senescent rough dog's tail grass contaminants might be moved to a neighbouring palatable pasture. The specific toxin(s) and their source, however, are still speculative, and it is uncertain if they are stable in the environment. The analysis of tissues from dead animals may thus be able to shed some light on this scenario, even if the concentration of the toxic substances that cause disease would be higher in feed source materials. Any unusual components may reveal information about the type of poison, according to a theory (s). Additionally, it is suspicious that there were many types of insects and regularly contaminated food present during an outbreak of liver injury. As a result, it is suggested that the source of the relevant toxin(s) may be one of numerous factors linked to increased liver damage in the cattle.

#### **Acknowledgements**

The authors thank the editors and anonymous publisher for their helpful feedback for this manuscript.

*The Focus on Core Genetic Factors That Regulate Hepatic Injury in Cattle Seems to Be… DOI: http://dx.doi.org/10.5772/intechopen.108151*

#### **Declaration of interest**

The authors declare that there is no conflict of interest for this manuscript.

#### **Informed consent statement**

Not applicable.

#### **Author details**

Avishek Mandal Department of Pharmaceutical Technology, Jadavpur University, Kolkata, India

\*Address all correspondence to: avishek7477@gmail.com

© 2022 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

#### **References**

[1] Zorov DB, Juhaszova M, Sollott SJ. Mitochondrial reactive oxygen species (ROS) and ROS-induced ROS release. Physiological Reviews. 2014;**94**(3):909- 950. DOI: 10.1152/physrev.00026. 2013

[2] Underwood WJ, Blauwiekel R, Delano ML, Gillesby R, Mischler SA, Schoell A. Biology and diseases of ruminants (sheep, goats, and cattle). Laboratory Animal Medicine. 2015;**11**:623-694. DOI: 10.1016/ B978-0-12-409527-4.00015-8

[3] Fabbrini E, Sullivan S, Klein S. Obesity and nonalcoholic fatty liver disease: Biochemical, metabolic, and clinical implications. Hepatology (Baltimore, Md.). 2010;**51**(2):679-689. DOI: 10.1002/hep.23280

[4] Savic D, Hodson L, Neubauer S, Pavlides M. The importance of the fatty acid transporter L-carnitine in nonalcoholic fatty liver disease (NAFLD). Nutrients. 2020;**12**(8):2178. DOI: 10.3390/nu12082178

[5] Sundrum A. Metabolic disorders in the transition period indicate that the dairy cows' ability to adapt is overstressed. Animals: An Open Access Journal from MDPI. 2015;**5**(4):978-1020. DOI: 10.3390/ani5040395

[6] Habel J, Chapoutot P, Koch C, Sundrum A. Estimation of individual glucose reserves in high-yielding dairy cows. Dairy. 2022;**3**(3):438-464

[7] Kuhla B. Review: Pro-inflammatory cytokines and hypothalamic inflammation: Implications for insufficient feed intake of transition dairy cows. Animal: An International Journal of Animal Bioscience.

2020;**14**(S1):s65-s77. DOI: 10.1017/ S1751731119003124

[8] Jain S, Gautam V, Naseem S. Acutephase proteins: As diagnostic tool. Journal of Pharmacy & Bioallied Sciences. 2011;**3**(1):118-127. DOI: 10.4103/0975-7406.76489

[9] Yuan K, Farney JK, Mamedova LK, Sordillo LM, Bradford BJ. TNFα altered inflammatory responses, impaired health and productivity, but did not affect glucose or lipid metabolism in early-lactation dairy cows. PLoS One. 2013;**8**(11):e80316. DOI: 10.1371/journal. pone.0080316

[10] Bossaert P, Trevisi E, Opsomer G, Bertoni G, De Vliegher S, Leroy J. The association between indicators of inflammation and liver variables during the transition period in high-yielding dairy cows: An observational study. Veterinary Journal (London, England : 1997). 2011;**192**:222-225. DOI: 10.1016/j. tvjl.2011.06.004

[11] Trevisi E, Minuti A. Assessment of the innate immune response in the periparturient cow. Research in Veterinary Science. 2017:**116**:116 DOI: 10.1016/j.rvsc.2017.12.001

[12] Rui L. Energy metabolism in the liver. Comprehensive Physiology. 2014;**4**(1):177-197. DOI: 10.1002/cphy. c130024

[13] Alves-Bezerra M, Cohen DE. Triglyceride metabolism in the liver. Comprehensive Physiology. 2017;**8**(1): 1-8. DOI: 10.1002/cphy.c170012

[14] Choi SH, Ginsberg HN. Increased very low density lipoprotein (VLDL) secretion, hepatic steatosis, and insulin *The Focus on Core Genetic Factors That Regulate Hepatic Injury in Cattle Seems to Be… DOI: http://dx.doi.org/10.5772/intechopen.108151*

resistance. Trends in Endocrinology and Metabolism: TEM. 2011;**22**(9):353-363. DOI: 10.1016/j.tem.2011.04.007

[15] Paschos P, Paletas K. Non alcoholic fatty liver disease and metabolic syndrome. Hippokratia. 2009;**13**(1):9-19

[16] Takahashi Y, Soejima Y, Fukusato T. Animal models of nonalcoholic fatty liver disease/nonalcoholic steatohepatitis. World Journal of Gastroenterology. 2012;**18**(19):2300-2308. DOI: 10.3748/ wjg.v18.i19.2300

[17] Simoes I, Janikiewicz J, Bauer J, Karkucinska-Wieckowska A, Kalinowski P, Dobrzyń A, et al. Fat and sugar-A dangerous duet. A comparative review on metabolic remodeling in rodent models of nonalcoholic fatty liver disease. Nutrients. 2019;**11**(12):2871. DOI: 10.3390/nu11122871

[18] Chen Z, Tian R, She Z, Cai J, Li H. Role of oxidative stress in the pathogenesis of nonalcoholic fatty liver disease. Free Radical Biology and Medicine. 2020;**152**:116-141

[19] Di Ciaula A, Passarella S, Shanmugam H, Noviello M, Bonfrate L, Wang DQ, et al. Nonalcoholic fatty liver disease (NAFLD). Mitochondria as players and targets of therapies? International Journal of Molecular Sciences. 2021;**22**(10):5375. DOI: 10.3390/ijms22105375

[20] Takaki A, Kawai D, Yamamoto K. Multiple hits, including oxidative stress, as pathogenesis and treatment target in non-alcoholic steatohepatitis (NASH). International Journal of Molecular Sciences. 2013;**14**(10):20704-20728. DOI: 10.3390/ijms141020704

[21] Mashek D. Hepatic lipid droplets: A balancing act between energy storage and metabolic dysfunction in NAFLD. Molecular Metabolism. 2021;**50**:101115

[22] Lin JH, Walter P, Yen TS. Endoplasmic reticulum stress in disease pathogenesis. Annual Review of Pathology. 2008;**3**:399-425. DOI: 10.1146/ annurev.pathmechdis.3.121806.151434

[23] Basseri S, Austin RC. Endoplasmic reticulum stress and lipid metabolism: Mechanisms and therapeutic potential. Biochemistry Research International. 2012;**2012**:841362. DOI: 10.1155/ 2012/841362

[24] Friedman SL, Neuschwander-Tetri BA, Rinella M, Sanyal AJ. Mechanisms of NAFLD development and therapeutic strategies. Nature Medicine. 2018;**24**(7):908-922. DOI: 10.1038/ s41591-018-0104-9

[25] Fujii J, Homma T, Kobayashi S, Seo H. Mutual interaction between oxidative stress and endoplasmic reticulum stress in the pathogenesis of diseases specifically focusing on non-alcoholic fatty liver disease. World Journal of Biological Chemistry. 2018;**9**:1-15. DOI: 10.4331/wjbc.v9.i1.1

[26] Benedict M, Zhang X. Non-alcoholic fatty liver disease: An expanded review. World Journal of Hepatology. 2017;**9**(16):715-732. DOI: 10.4254/wjh. v9.i16.715

[27] Tilg H, Moschen A. Insulin resistance, inflammation, and nonalcoholic fatty liver disease. Trends in Endocrinology and Metabolism: TEM. 2008;**19**:371-379. DOI: 10.1016/j. tem.2008.08.005

[28] Hussain MM, Nijstad N, Franceschini L. Regulation of microsomal triglyceride transfer protein. Clinical Lipidology. 2011;**6**(3):293-303. DOI: 10.2217/clp.11.21

[29] Tietge UJ, Bakillah A, Maugeais C, Tsukamoto K, Hussain M, Rader DJ.

Hepatic overexpression of microsomal triglyceride transfer protein (MTP) results in increased in vivo secretion of VLDL triglycerides and apolipoprotein B. Journal of Lipid Research. 1999;**40**(11):2134-2139

[30] Hocquette J, Bauchart D. Intestinal absorption, blood transport and hepatic and muscle metabolism of fatty acids in preruminant and ruminant animals. Reproduction Nutrition Development. 1999;**39**(1):27-48

[31] Améen C, Edvardsson U, Ljungberg A, Asp L, Akerblad P, Tuneld A, et al. Activation of peroxisome proliferator-activated receptor increases the expression and activity of microsomal triglyceride transfer protein in the liver. The Journal of Biological Chemistry. 2005;**280**:1224-1229. DOI: 10.1074/jbc.M412107200

[32] Bakillah A, Hussain MM. Mice subjected to *aP2-Cre* mediated ablation of microsomal triglyceride transfer protein are resistant to high fat diet induced obesity. Nutrition & Metabolism (London). 2016;**13**:1. DOI: 10.1186/ s12986-016-0061-6

[33] Kersten S. Peroxisome proliferator activated receptors and lipoprotein metabolism. PPAR Research. 2008;**2008**:1-11

[34] Trajkovski M, Hausser J, Soutschek J, Bhat B, Akin A, Zavolan M, et al. MicroRNAs 103 and 107 regulate insulin sensitivity. Nature. 2011;**474**(7353):649-653. DOI: 10.1038/ nature10112

[35] Rau CS, Wu SC, Lu TH, Wu YC, Wu CJ, Chien PC, et al. Effect of lowfat diet in obese mice lacking toll-like receptors. Nutrients. 2018;**10**(10):1464. DOI: 10.3390/nu10101464

[36] Foley N, O'Neill L. miR-107: A toll-like receptor-regulated miRNA dysregulated in obesity and type II diabetes. Journal of Leukocyte Biology. 2012;**92**:521-527. DOI: 10.1189/ jlb.0312160

[37] Borrelli A, Bonelli P, Tuccillo FM, Goldfine ID, Evans JL, Buonaguro FM, et al. Role of gut microbiota and oxidative stress in the progression of non-alcoholic fatty liver disease to hepatocarcinoma: Current and innovative therapeutic approaches. Redox Biology. 2018;**15**:467-479. DOI: 10.1016/j.redox.2018.01.009

[38] Penn JS, Madan A, Caldwell RB, Bartoli M, Caldwell RW, Hartnett ME. Vascular endothelial growth factor in eye disease. Progress in Retinal and Eye Research. 2008;**27**(4):331-371. DOI: 10.1016/j.preteyeres.2008.05.001

[39] Ferrara N. Role of vascular endothelial growth factor in the regulation of angiogenesis. Kidney International. 1999;**56**(3):794-814

[40] Shibuya M. Vascular endothelial growth factor (VEGF) and its receptor (VEGFR) signaling in angiogenesis: A crucial target for anti- and pro-Angiogenic therapies. Genes & Cancer. 2011;**2**(12):1097-1105. DOI: 10.1177/ 1947601911423031

[41] Rahimi N. VEGFR-1 and VEGFR-2: Two non-identical twins with a unique physiognomy. Frontiers in Bioscience: a Journal and Virtual Library. 2006;**11**:818- 829. DOI: 10.2741/1839

[42] Kaufman N, Dhingra S, Jois S, Vicente M. Molecular targeting of epidermal growth factor receptor (EGFR) and vascular endothelial growth factor receptor (VEGFR). Molecules. 2021;**26**(4):1076

*The Focus on Core Genetic Factors That Regulate Hepatic Injury in Cattle Seems to Be… DOI: http://dx.doi.org/10.5772/intechopen.108151*

[43] Li YL, Zhao H, Ren XB. Relationship of VEGF/VEGFR with immune and cancer cells: Staggering or forward? Cancer Biology & Medicine. 2016;**13**(2):206-214. DOI: 10.20892/j. issn.2095-3941.2015.0070

[44] Spencer TE. Biological roles of uterine glands in pregnancy. Seminars in Reproductive Medicine. 2014;**32**(5):346-357. DOI: 10.1055/ s-0034-1376354

[45] Pfarrer C, Ruziwa S, Winther H, Callesen H, Leiser R, Schams D, et al. Localization of vascular endothelial growth factor (Vegf) and its receptors Vegfr-1 and Vegfr-2 In bovine Placentomes from implantation until term. Placenta. 2006;**27**(8):889-898

[46] Chiumia D, Hankele A-K, Groebner A, Schulke K, Reichenbach H-D, Giller K, et al. Vascular endothelial growth factor A and VEGFR-1 change during preimplantation in heifers. International Journal of Molecular Sciences. 2020;**21**:544. DOI: 10.3390/ ijms21020544

[47] Carvajal L, Gutiérrez J, Morselli E, Leiva A. Autophagy process in trophoblast cells invasion and differentiation: Similitude and differences with cancer cells. Frontiers in Oncology. 2021;**11**:11

[48] Kucera O, Cervinkova Z. Experimental models of non-alcoholic fatty liver disease in rats. World Journal of Gastroenterology. 2014;**20**(26):8364- 8376. DOI: 10.3748/wjg.v20.i26.8364

[49] Lei L, Ei Mourabit H, Housset C, Cadoret A, Lemoinne S. Role of angiogenesis in the pathogenesis of NAFLD. Journal of Clinical Medicine. 2021;**10**(7):1338. DOI: 10.3390/ jcm10071338

[50] Li Q, Dhyani M, Grajo JR, Sirlin C, Samir AE. Current status of imaging in nonalcoholic fatty liver disease. World Journal of Hepatology. 2018;**10**(8):530- 542. DOI: 10.4254/wjh.v10.i8.530

[51] Ofoedu CE, Iwouno JO, Ofoedu EO, Ogueke CC, Igwe VS, Agunwah IM, et al. Revisiting food-sourced vitamins for consumer diet and health needs: A perspective review, from vitamin classification, metabolic functions, absorption, utilization, to balancing nutritional requirements. PeerJ. 2021;**9**:e11940. DOI: 10.7717/peerj.11940

[52] Moghadaszadeh B, Beggs AH. Selenoproteins and their impact on human health through diverse physiological pathways. Physiology (Bethesda, Md.). 2006;**21**:307-315. DOI: 10.1152/physiol.00021.2006

[53] Surai PF, Kochish II, Fisinin VI, Juniper DT. Revisiting oxidative stress and the use of organic selenium in dairy cow nutrition. Animals: An Open Access Journal from MDPI. 2019;**9**(7):462. DOI: 10.3390/ani9070462

[54] Abuelo A, Hernández J, Benedito J, Castillo C. Redox biology in transition periods of dairy cattle: Role in the health of Periparturient and neonatal. Animals. 2019;**8**:20. DOI: 10.3390/antiox8010020

[55] Traber MG. Vitamin E inadequacy in humans: Causes and consequences. Advances in nutrition (Bethesda, Md.). 2014;**5**(5):503-514. DOI: 10.3945/ an.114.006254

[56] Deters E, Hansen S. Invited review: Linking road transportation with oxidative stress in cattle and other species. Applied Animal Science. 2020;**36**(2):183-200

[57] Richeson J, Falkner T. Bovine respiratory disease vaccination.

Veterinary Clinics of North America: Food Animal Practice. 2020;**36**:473-485. DOI: 10.1016/j.cvfa.2020.03.013

[58] Wilson BK, Richards CJ, Step DL, Krehbiel CR. Best management practices for newly weaned calves for improved health and well-being. Journal of Animal Science. 2017;**95**(5):2170-2182. DOI: 10.2527/jas.2016.1006

[59] Ponnampalam E, Hopkins D, Bruce H, Baldi G, Bekhit A. Causes and contributing factors to "dark cutting" meat: Current trends and future directions: A review. Comprehensive Reviews in Food Science and Food Safety. 2017;**16**:16. DOI: 10.1111/1541-4337.12258

[60] Moussa Z, Judeh ZM, Ahmed SA. Nonenzymatic exogenous and endogenous antioxidants. Free Radical Medicine and Biology. 2020;**1** DOI: 10.5772/intechopen.87778

[61] Ma Q. Role of nrf2 in oxidative stress and toxicity. Annual Review of Pharmacology and Toxicology. 2013;**53**:401-426. DOI: 10.1146/ annurev-pharmtox-011112-140320

[62] Zhong Q, Mishra M, Kowluru R. Transcription factor Nrf2-mediated antioxidant defense system in the development of diabetic retinopathy. Investigative Opthalmology & Visual Science. 2013;**54**(6):3941

[63] Melnik BC, Schmitz G. Milk's role as an epigenetic regulator in health and disease. Diseases (Basel, Switzerland). 2017;**5**(1):12. DOI: 10.3390/ diseases5010012

[64] Aydin Y, Chedid M, Chava S, Danielle Williams D, Liu S, Hagedorn CH, et al. Activation of PERK-Nrf2 oncogenic signaling promotes Mdm2-mediated Rb degradation in persistently infected HCV culture.

Scientific Reports. 2017;**7**(1):9223. DOI: 10.1038/s41598-017-10087-6. (Retraction published Sci Rep. 2021 Sep 1;11(1):17779)

[65] Mittal M, Siddiqui MR, Tran K, Reddy SP, Malik AB. Reactive oxygen species in inflammation and tissue injury. Antioxidants & Redox Signaling. 2014;**20**(7):1126-1167. DOI: 10.1089/ ars.2012.5149

[66] Tan BL, Norhaizan ME, Liew WP, Sulaiman Rahman H. Antioxidant and oxidative stress: A mutual interplay in age-related diseases. Frontiers in Pharmacology. 2018;**9**:1162. DOI: 10.3389/fphar.2018.01162

[67] Kurutas EB. The importance of antioxidants which play the role in cellular response against oxidative/ nitrosative stress: Current state. Nutrition Journal. 2015;**15**:71. DOI: 10.1186/s12937-016-0186-5

[68] Judge A, Dodd MS. Metabolism. Essays in Biochemistry. 2020;**64**(4): 607-647. DOI: 10.1042/EBC20190041

[69] Marchi S, Giorgi C, Suski J, Agnoletto C, Bononi A, Bonora M, et al. Mitochondria-Ros crosstalk in the control of cell death and aging. Journal of Signal Transduction. 2012;**2012**:1-17

[70] Videla L, Rodrigo R, Araya J, Poniachik J. Insulin resistance and oxidative stress interdependency in non-alcoholic fatty liver disease. Trends in Molecular Medicine. 2007;**12**:555-558. DOI: 10.1016/j.molmed.2006.10.001

[71] Arroyave-Ospina JC, Wu Z, Geng Y, Moshage H. Role of oxidative stress in the pathogenesis of non-alcoholic fatty liver disease: Implications for prevention and therapy. Antioxidants (Basel, Switzerland). 2021;**10**(2):174. DOI: 10.3390/antiox10020174

*The Focus on Core Genetic Factors That Regulate Hepatic Injury in Cattle Seems to Be… DOI: http://dx.doi.org/10.5772/intechopen.108151*

[72] Harjumäki R, Pridgeon CS, Ingelman-Sundberg M. CYP2E1 in alcoholic and non-alcoholic liver injury. Roles of ROS, reactive intermediates and lipid overload. International Journal of Molecular Sciences. 2021;**22**(15):8221. DOI: 10.3390/ijms22158221

[73] Liput KP, Lepczyński A, Ogłuszka M, Nawrocka A, Poławska E, Grzesiak A, et al. Effects of dietary n-3 and n-6 polyunsaturated fatty acids in inflammation and Cancerogenesis. International Journal of Molecular Sciences. 2021;**22**(13):6965. DOI: 10.3390/ijms22136965

[74] Tangvarasittichai S. Oxidative stress, insulin resistance, dyslipidemia and type 2 diabetes mellitus. World Journal of Diabetes. 2015;**6**(3):456-480. DOI: 10.4239/wjd.v6.i3.456

[75] Ma Y, Lee G, Heo SY, Roh YS. Oxidative stress is a key modulator in the development of nonalcoholic fatty liver disease. Antioxidants (Basel, Switzerland). 2021;**11**(1):91. DOI: 10.3390/antiox11010091

[76] Di Meo S, Reed TT, Venditti P, Victor VM. Role of ROS and RNS sources in physiological and pathological conditions. Oxidative Medicine and Cellular Longevity. 2016;**2016**:1245049. DOI: 10.1155/2016/1245049

[77] Bhattacharyya A, Chattopadhyay R, Mitra S, Crowe SE. Oxidative stress: An essential factor in the pathogenesis of gastrointestinal mucosal diseases. Physiological Reviews. 2014;**94**(2):329- 354. DOI: 10.1152/physrev.00040.2012

[78] Chawengsub Y, Gauthier KM, Campbell WB. Role of arachidonic acid lipoxygenase metabolites in the regulation of vascular tone. American Journal of Physiology. Heart and Circulatory Physiology.

2009;**297**(2):H495-H507. DOI: 10.1152/ ajpheart.00349.2009

[79] McFadden J. Review: Lipid Biology in the Periparturient Dairy Cow: Animal Contemporary Perspectives. 2022;**14**

[80] Strickland J, Wisnieski L, Mavangira V, Sordillo L. Serum vitamin D is associated with antioxidant potential in Peri-parturient cows. Antioxidants. 2021;**10**(9):1420

[81] Ito F, Sono Y, Ito T. Measurement and clinical significance of lipid peroxidation as a biomarker of oxidative stress: Oxidative stress in diabetes, atherosclerosis, and chronic inflammation. Antioxidants (Basel, Switzerland). 2019;**8**(3):72. DOI: 10.3390/antiox8030072

[82] Mavangira V, Gandy J, Zhang C, Ryman V, Jones A, Sordillo L. Polyunsaturated fatty acids influence differential biosynthesis of oxylipids and other lipid mediators during bovine coliform mastitis. Journal of Dairy Science. 2015;**98**:1-14. DOI: 10.3168/ jds.2015-9570

[83] Mattmiller S, Carlson B, Gandy J, Sordillo L. Reduced macrophage selenoprotein expression alters oxidized lipid metabolite biosynthesis from arachidonic and linoleic acid. The Journal of Nutritional Biochemistry. 2014;**25**: 1-16. DOI: 10.1016/j.jnutbio.2014.02.005

[84] Silva L, dos Santos Neto A, Maia S, dos Santos Guimarães C, Quidute I, Carvalho A, et al. The role of TNF-α as a Proinflammatory cytokine in pathological processes. The Open Dentistry Journal. 2019;**13**(1):332-338

[85] Yamada Y, Kirillova I, Peschon JJ, Fausto N. Initiation of liver growth by tumor necrosis factor: Deficient liver regeneration in mice lacking type I tumor necrosis factor receptor. Proceedings of the National Academy of Sciences of the United States of America. 1997;**94**(4):1441-1446. DOI: 10.1073/ pnas.94.4.1441

[86] Jaffer U, Wade RG, Gourlay T. Cytokines in the systemic inflammatory response syndrome: A review. HSR Proceedings in Intensive Care & Cardiovascular Anesthesia. 2010;**2**(3):161-175

[87] Wheelock JB, Rhoads RP, Vanbaale MJ, Sanders SR, Baumgard LH. Effects of heat stress on energetic metabolism in lactating Holstein cows. Journal of Dairy Science. 2010;**93**(2):644-655. DOI: 10.3168/ jds.2009-2295

[88] Hassan FU, Nadeem A, Javed M, Saif-Ur-Rehman M, Shahzad MA, Azhar J, et al. Nutrigenomic interventions to address metabolic stress and related disorders in transition cows. BioMed Research International. 2022;**2022**:2295017. DOI: 10.1155/2022/ 2295017

[89] de Boer G, Robinson PH, Kennelly JJ. Hormonal responses to bovine somatotropin and dietary protein in early lactation dairy cows. Journal of Dairy Science. 1991;**74**(8):2623-2632. DOI: 10.3168/jds.S0022-0302(91)78441-5

[90] Kushibiki S, Hodate K, Shingu H, Obara Y, Touno E, Shinoda M, et al. Metabolic and Lactational responses during recombinant bovine tumor necrosis factor-α treatment in lactating cows. Journal of Dairy Science. 2003;**86**(3):819-827

[91] Chait A, den Hartigh LJ. Adipose tissue distribution, inflammation and its metabolic consequences, including diabetes and cardiovascular disease. Frontiers in Cardiovascular

Medicine. 2020;**7**:22. DOI: 10.3389/ fcvm.2020.00022

[92] Lima R, Nunes P, Viana A, Oliveira F, Silva R, Alves A, et al. α,β-Amyrin prevents steatosis and insulin resistance in a high-fat diet-induced mouse model of NAFLD via the AMPKmTORC1-SREBP1 signaling mechanism. Brazilian Journal of Medical and Biological Research. 2021;**54**(10)

[93] Mezzetti M, Bionaz M, Trevisi E. Interaction between inflammation and metabolism in periparturient dairy cows. Journal of Animal Science. 2020;**98**(Suppl 1):1-9. DOI: 10.1093/jas/ skaa134

[94] Kushibiki S, Shingu H, Komatsu T, Itoh F, Kasuya E, Aso H, et al. Effect of recombinant tumor necrosis factor-α on hormone release in lactating cows. Animal Science Journal. 2006;**77**:603- 612. DOI: 10.1111/j.1740-0929. 2006.00392.x

[95] Abouelasrar Salama S, De Bondt M, De Buck M, Berghmans N, Proost P, Oliveira V, et al. Serum amyloid A1 (SAA1) revisited: Restricted leukocyte-activating properties of homogeneous SAA1. Frontiers in Immunology. 2020;**11**:11

[96] Yang YM, Seki E. TNFα in liver fibrosis. Current Pathobiology Reports. 2015;**3**(4):253-261. DOI: 10.1007/ s40139-015-0093-z

[97] Tao S, Dahl G. Invited review: Heat stress effects during late gestation on dry cows and their calves. Journal of Dairy Science. 2013;**96**(7):4079-4093

[98] Loor J, Dann H, Everts R, Oliveira R, Green C, Guretzky N, et al. Temporal gene expression profiling of liver from periparturient dairy cows reveals complex adaptive mechanisms in hepatic *The Focus on Core Genetic Factors That Regulate Hepatic Injury in Cattle Seems to Be… DOI: http://dx.doi.org/10.5772/intechopen.108151*

function. Physiological Genomics. 2005;**23**(2):217-226

[99] Thering BJ, Bionaz M, Loor J. Longchain fatty acid effects on peroxisome proliferator-activated receptor-αregulated genes in Madin-Darby bovine kidney cells: Optimization of culture conditions using palmitate. Journal of Dairy Science. 2009;**92**:2027-2037. DOI: 10.3168/jds.2008-1749

[100] Contreras AV, Rangel-Escareño C, Torres N, Alemán-Escondrillas G, Ortiz V, Noriega LG, et al. PPARα via HNF4α regulates the expression of genes encoding hepatic amino acid catabolizing enzymes to maintain metabolic homeostasis. Genes & Nutrition. 2015;**10**(2):452. DOI: 10.1007/ s12263-014-0452-0

[101] Xu H, Luo J, Zhao W, Yang Y, Tian H, Shi H, et al. Overexpression of SREBP1 (sterol regulatory element binding protein 1) promotes de novo fatty acid synthesis and triacylglycerol accumulation in goat mammary epithelial cells. Journal of Dairy Science. 2016;**99**(1):783-795

[102] Ress C, Kaser S. Mechanisms of intrahepatic triglyceride accumulation. World Journal of Gastroenterology. 2016;**22**(4):1664-1673. DOI: 10.3748/wjg. v22.i4.1664

[103] Brismar K, Fernqvist-Forbes E, Wahren J, Hall K. Effect of insulin on the hepatic production of insulin-like growth factor-binding protein-1 (IGFBP-1), IGFBP-3, and IGF-I in insulindependent diabetes. The Journal of Clinical Endocrinology and Metabolism. 1994;**79**(3):872-878. DOI: 10.1210/ jcem.79.3.7521354

[104] Ijuin T, Takenawa T. Regulation of insulin signaling by the phosphatidylinositol 3,4,5-triphosphate phosphatase SKIP through the scaffolding function of Pak1. Molecular and Cellular Biology. 2012;**32**(17):3570- 3584. DOI: 10.1128/MCB.00636-12

[105] Saha S, Buttari B, Panieri E, Profumo E, Saso L. An overview of Nrf2 signaling pathway and its role in inflammation. Molecules (Basel, Switzerland). 2020;**25**(22):5474. DOI: 10.3390/molecules25225474

[106] Deshmukh P, Unni S, Krishnappa G, Padmanabhan B. The Keap1-Nrf2 pathway: Promising therapeutic target to counteract ROS-mediated damage in cancers and neurodegenerative diseases. Biophysical Reviews. 2017;**9**(1):41-56. DOI: 10.1007/ s12551-016-0244-4

[107] Sharifi-Rad M, Anil Kumar N, Zucca P, Varoni E, Dini L, Panzarini E, et al. Lifestyle, oxidative stress, and antioxidants: Back and forth in the pathophysiology of chronic diseases. Frontiers in Physiology. 2020;**11**:11

[108] Xiong L, Xie J, Song C, Liu J, Zheng J, Liu C, et al. The activation of Nrf2 and its downstream regulated genes mediates the Antioxidative activities of Xueshuan Xinmaining tablet in human umbilical vein endothelial cells. Evidence-Based Complementary and Alternative Medicine: eCAM. 2015;**2015**:187265. DOI: 10.1155/ 2015/187265

[109] Ringseis R, Gessner D, Eder K. Molecular insights into the mechanisms of liver-associated diseases in early-lactating dairy cows: Hypothetical role of endoplasmic reticulum stress. Journal of Animal Physiology and Animal Nutrition. 2014;**99**(4):626-645

[110] Miyake K. Innate immune sensing of pathogens and danger signals

by cell surface toll-like receptors. SeminImmunol. 2007;**19**:3-10

[111] Medzhitov R, Preston-Hurlburt P, Janeway CA. A human homologue of the drosophila toll protein signals activation of adaptive immunity. Nature. 1997;**388**:394-397

[112] Akira S, Takeda K, Kaisho T. Tolllike receptors: Critical proteins linking innate and acquired immunity. Nature Immunology. 2007;**2**:675-680

[113] Kuhn H, Petzold K, Hammerschmidt S, Wirtz H. Interaction of cyclic mechanical stretch and toll-like receptor4-mediated innate immunity in rat alveolar type cells. Respirology. 2014;**19**:67-73. DOI: 10.1111/resp.12149

[114] Yang J, Yang J, Ding JW, Chen LH, Wang YL, Li S, et al. Sequential expression of TLR4 and its effects on the myocardium of rats with myocardial ischemia-reperfusion injury. Inflammation. 2008;**31**:304-312. DOI: 10.1007/s10753-008-9079-x

[115] O'Neill LA, Bowie AG. The family of five: TIR-domain-containing adaptors in toll-like receptor signalling. Nature Reviews. Immunology. 2007;**7**:353-364

[116] Fitzgerald KA, Palsson-McDermott EM, Bowie AG, Jefferies CA, Mansell AS, Brady G, et al. Mal (MyD88 adapter-like) is required for toll-like receptor-4 signal transduction. Nature. 2001;**413**:78-83

[117] Gilbert FB, Cunha P, Jensen K, Glass EJ, Foucras G, Robert-Granié C, et al. Differential response of bovine mammary epithelial cells to Staphylococcus aureus or Escherichia coli agonists of the innate immune system. Veterinary Research. 2013;**44**:40. DOI: 10.1186/1297-9716-44-40

[118] Sipka A, Klaessig S, Duhamel GE, Swinkels J, Rainard P, Schukken Y. Impact of Intramammary treatment on gene expression profiles in bovine Escherichia coli mastitis. PLoS One. 2014;**9**(1):e85579. DOI: 10.1371/journal. pone.0085579

[119] Blasi E, Ardizzoni A, Colombari B, Neglia R, Baschieri C, Peppoloni S, et al. NF-κB activation and p38 phosphorylation in microglial cells infected with Leptospira or exposed to partially purified leptospiral lipoproteins. Microbial Pathogenesis. 2007;**42**:80-87

[120] Liu SL, Kielian T. Microglial activation by Citrobacter koseri is mediated by TLR4-and MyD88 dependent pathways. Journal of Immunology. 2009;**183**:5537-5547. DOI: 10.4049/jimmunol.0900083

[121] Wei Z, Zhu NS, Bai D, Miao JF, Zou SX. The crosstalk between Dectin1 and TLR4 via NF-κB subunits p65/ RelB in mammary epithelial cells. International Immunopharmacology; **23** DOI: 10.1016/j.intimp.2014;09.004

[122] Wei ZK, Zhou ES, Guo CM, Fu YH, Yu YQ, Li YM, et al. Thymol inhibits Staphylococcus aureus internalization into bovine mammary epithelial cells by inhibiting NF-kB activation. Microbial Pathogenesis. 2014;**11**:1-5

[123] Martina JA, Diab HI, Lishu L, Jeong-A L, Patange S, Raben N, et al. The nutrient-responsive transcription factor TFE3 promotes autophagy, lysosomal biogenesis, and clearance of cellular debris. Science Signaling. 2014;**7**:ra9. DOI: 10.1126/scisignal.2004754

[124] Yin Q, Jian Y, Xu M, Huang X, Wang N, Liu Z, et al. CDK4/6 regulate lysosome biogenesis through TFEB/ TFE3. The Journal of Cell Biology.

*The Focus on Core Genetic Factors That Regulate Hepatic Injury in Cattle Seems to Be… DOI: http://dx.doi.org/10.5772/intechopen.108151*

2020;**219**:e201911036. DOI: 10.1083/ jcb.201911036

[125] Settembre C, De Cegli R, Mansueto G, Saha PK, Vetrini F, Visvikis O, et al. TFEB controls cellular lipid metabolism through a starvationinduced autoregulatory loop. Nature Cell Biology. 2013;**15**:647-658. DOI: 10.1038/ ncb2718

[126] Settembre C, Di Malta C, Polito VA, Garcia Arencibia M, Vetrini F, Erdin S, et al. TFEB links autophagy to lysosomal biogenesis. Science. 2011;**332**:1429-1433. DOI: 10.1126/science.1204592

[127] Sinha RA, Farah BL, Singh BK, Siddique MM, Li Y, Wu Y, et al. Caffeine stimulates hepatic lipid metabolism by the autophagylysosomal pathway in mice. Hepatology. 2014;**59**:1366-1380

[128] Shen T, Li X, Loor JJ, Zhu Y, Du X, Wang X, et al. Hepatic nuclear factor kappa B signaling pathway and NLR family pyrin domain containing 3 inflammasome is over-activated in ketotic dairy cows. Journal of Dairy Science. 2019;**102**:10554-10563. DOI: 10.3168/jds.2019-16706

[129] Napolitano G, Di Malta C, Esposito A, de Araujo MEG, Pece S, Bertalot G, et al. A substrate-specific mTORC1 pathway underlies Birt-Hogg-Dubé syndrome. Nature. 2020;**585**:597- 602. DOI: 10.1038/s41586-020-2444-0

[130] Palmieri M, Pal R, Nelvagal HR, Lotfi P, Stinnett GR, Seymour ML, et al. mTORC1-independent TFEB activation via Akt inhibition promotes cellular clearance in neurodegenerative storage diseases. Nature Communications. 2017;**8**:14338. DOI: 10.1038/ ncomms14338

[131] Li X, Shi Z, Zhu Y, Shen T, Wang H, Shui G, et al. Cyanidin-3-O-glucoside

improves non-alcoholic fatty liver disease by promoting PINK1-mediated mitophagy in mice. British Journal of Pharmacology. 2020;**177**:3591-3607. DOI: 10.1111/bph.15083

[132] Zachut M, Honig H, Striem S, Zick Y, Boura-Halfon S, Moallem U. Periparturient dairy cows do not exhibit hepatic insulin resistance, yet adiposespecific insulin resistance occurs in cows prone to high weight loss. Journal of Dairy Science. 2013;**96**:5656-5669. DOI: 10.3168/jds.2012-6142

[133] Laplante M, Sabatini DM. mTOR signaling in growth control and disease. Cell. 2012;**149**:274-293. DOI: 10.1016/j. cell.2012.03.017

[134] Hou B, Li Y, Li X, Zhang C, Zhao Z, Chen Q, et al. HGF protected against diabetic nephropathy via autophagylysosome pathway in podocyte by modulating PI3K/ Akt-GSK3β-TFEB axis. Cellular Signalling. 2020;**75**:109744. DOI: 10.1016/j.cellsig.2020.109744
