**5. Molecular mechanisms of PH-induced LPO**

*Acridocarpus smeathmannii* (DC.) is widely used for the treatment of male infertility, anemia, and pains in traditional medicine as well as an herbal mixture. *A. smeathmannii* has gained popularity particularly worldwide for the management of infertility, anemia, pain, and some cutaneous infections [20, 41]. Studies have demonstrated the ethnobotanical relevance of this plant. Thus, recent reports demonstrated the bioactive compounds, sexual behavior, and associated reproductive function via biochemical and pharmacological mechanisms due to the hydroethanolic root extract of *A. smethmannii* (HAS) and subchronic oral toxicity effects of HAS [31]. Also, the organization for economic co-operation and development (Test No. 453) [39] has approved that very long-term toxicological and/or carcinogenicity studies be carried, in particular for PH that are considered not to be potentially toxic at therapeutic and supratherapeutic doses. In this study, the possible systemic toxicological changes following a 180 days administration in Wistar rats of both sexes under approved guidelines for animal use following the procedures as documented by Kilkenny et al. [42] for reporting animal research. Animals (Wistar rats, male: female = 1:1 = 48) received distilled water (10 mL/kg) or HAS (250, 500 or 1000 mg/kg body weight per day) consecutively for 180 days. From the results obtained, HAS (500 and 1000 mg/kg) demonstrates such a tendency to increase oxidative stress parameters via an increase in LPO as malondialdehyde (MDA) levels in vital organs (**Figure 1**) in rats. Although, evidence abounds of the turnover of products of LPO in the body, however, the presence of an adverse reaction may become aggravated time-dependently or produces an interaction with other substances presences [14, 26]. This could impose either a self or even exaggerates the effect of HAS in a given period. Increasing levels of the intracellular antioxidants, GSH levels, could not overcome the LPO products induced by the highest dose of extract in rats, thus, highlighting potential adverse effects of HAS in vivo. This suggests that PH can induce LPO metabolites such as MDA which by this observation could translate into a clinically relevant situation. More so, oestrogen level was reduced in treated female rats as obtained in these results. Also, in both sexes, HAS (500 mg/kg) and HAS (1000 mg/kg) showed elevated serum nitric oxide (NO) levels respectively (**Table 2**).

#### **Figure 1.**

*Effect of HAS on lipid peroxidation in normal Wistar rats. Results are expressed as mean ± S.E.M. n: Total number per group. n = 12. Mortality = HAS (500) (male, 25%), HAS (1000) (male, 33.3%), HAS1 (250 mg/kg) (female, 8.3%), HAS2 (500 mg/kg) (female, 25%) and HAS (1000 mg/kg) (female, 41.7%) respectively. \* p < 0.05 or \*\*p < 0.01 compared with control (distilled water: DW, 10 mL/kg) group. HAS1 (250): 250 mg/kg, HAS2 (500): 500 mg/kg, HAS3 (1000): 1000 mg/kg, HAS: Hydroethanolic extract of*  Acridocarpus smeathmannii *root.*


*Results are expressed as mean ± S.E.M. n = 12. HAS: hydroethanolic extract of Acridocarpus smeathmannii root. \* p < 0.05 or \*\*p < 0.01 compared with control distilled water group. "x" and "y" in superscript represented "male" and "female" rats respectively. Mortality: HAS (500) (male, 25%), HAS (1000) (male, 33.3%), HAS (250) ( female, 8.3%), HAS (500) ( female, 25%) and HAS (1000) ( female, 41.7%). Oestrogen (pg/mL), Progesterone (ng/mL), Testosterone (ng/mL), Serum Nitric Oxide (nmol/mL), TNF-*α*: Tumor Necrosis Factor-alpha, PSA: Prostate Specific Antigen (ng/mL). Differences between groups were determined by one-way analysis of variance (ANOVA) using Statistical Package for Social Sciences (SPSS, version 20.0) software for windows and Post hoc test for intergroup using the least significant difference, followed by Dunnett's test. Significance was considered at p < 0.05. All results were expressed as the mean ± standard error of the mean.*

#### **Table 2.**

*Effect of hydroethanolic root of* Acridocarpus smeathmannii *extract on body hormones and molecular biomarkers in serum of normal and treated rats using enzyme-linked immunosorbent assays.*

#### *Lipid Peroxidation and the Redox Effects of Polyherbal DOI: http://dx.doi.org/10.5772/intechopen.97625*

Nitric oxide plays a very significant role as a signaling molecule in several biological events [43]. Also, the highest dose, HAS (1000 mg/kg) increased serum tumor necrosis factor-alpha (TNF-α) level, a hallmark biomarker of tissue injury, in rats of both sexes (**Table 2**). The presence of TNF-α level could indicate some processes involving the action of nuclear factor kappa-B and a host of other mediators of injury [44]. Even at the lowest dose (250 mg/kg) of HAS administered to rats, serum NF-kB levels increase (**Table 2**). The aforementioned provides such suitability that HAS administration may induce cellular changes ranging from inflammatory reactions to LPO metabolites. An inducible nitric oxide synthase (iNOS) is an importantly upregulated isoform of NOS that synthesis the inducible nitric oxide (iNO) [45]. iNO is a disease-causing agent that can increase iNOS mRNA, protein, and then causes activity that may significantly alter NO turnover [42]. From the study results, administration of HAS after 180 days increases the chances of iNOS expression (**Figure 2**) in rats thereby showing the potential inherent pro-inflammatory and pro-oxidant properties in HAS that can cause tissue damage.
