**Traditional Chinese Medicine Can Improve Liver Microcirculation and Reduce Portal Hypertension in Liver Cirrhosis**

Xu Lieming1,5, Gu Jie2, Lu Xiong2, Zhou Yang1,5, Tian Tian2, Zhang Jie2 and Xu Hong2 *1Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, 2Institute of Liver Diseases, Shanghai University of TCM 3Key Laboratory of Liver and Kidney Diseases, Ministry of Education 4Key Laboratory of Traditional Chinese Medicine Clinic, Shanghai 5The Key Unit of Liver Diseases, SATCM. Shanghai, China* 

#### **1. Introduction**

92 Portal Hypertension – Causes and Complications

Cárdenas, A. & Ginès, P. (2009). Portal hypertension. *Curr Opin Gastroenterol,* Vol.25, №3,

Choi, Y.J.; Baik, S.K.; Park, D.H.; Kim, M.Y.; Kim, H.S.; Lee, D.K.; Kwon, S.O.; Kim, Y.J. &

Coelho-Prabhu, N. & Kamath, P.S. (2010). Current staging and diagnosis of gastroesophageal varices. *Clin Liver Dis,* Vol.14, №2, (May 2010), pp. 195-208 de Franchis, R.; Eisen, G.M.; Laine, L.; Fernandez-Urien, I.; Herrerias, J.M.; Brown, R.D.;

with portal hypertension. *Hepatology,* Vol.47, №5, (May 2008), pp. 1595-1603 de Franchis, R. & Baveno V Faculty (2010). Revising consensus in portal hypertension:

Park, J.W. (2003). Comparison of Doppler ultrasonography and the hepatic venous pressure gradient in assessing portal hypertension in liver cirrhosis. *J Gastroenterol* 

Fisher, L.; Vargas, H.E.; Vargo, J.; Thompson, J. & Eliakim, R. (2008). Esophageal capsule endoscopy for screening and surveillance of esophageal varices in patients

report of the Baveno V consensus workshop on methodology of diagnosis and therapy in portal hypertension. *J Hepatol,* Vol.53, №4, (October 2010), pp. 762-768 Groszmann, R.; Vorobioff, J.D. & Gao, H. (2006). Measurement of portal pressure: when, how, and why to do it. *Clin Liver Dis,* Vol.10, №3, (August 2006), pp. 499-512 Japanese Research Society for Portal Hypertension (1980). The general rules for recording

endoscopic findings on esophageal varices. *Jpn J Surg,* Vol.10, №1, (March 1980),

Kroupa, R. (2005). Variceal bleeding in portal hypertension: bacterial infection and comparison of efficacy of intravenous and per-oral application of antibiotics - a randomized trial. *Eur J Gastroenterol Hepatol,* Vol.17, №10, (October 2005), pp. 1105-

(2010). Increased plasma malondialdehyde in patients with viral cirrhosis and its relationships to plasma nitric oxide, endotoxin, and portal pressure. *Dig Dis Sci,*

coagulation by endotoxin. *Thromb Diath Haemorrh,* Vol.19, №1, (March 1968), pp.

S.S. & Lo, K.J. (1995). Endotoxemia in patients with chronic liver diseases: relationship to severity of liver diseases, presence of esophageal varices, and hyperdynamic circulation. *J Hepatol,* Vol.22, №2, (February 1995), pp. 165-172 Pugh, R.N.H.; Murray-Lyon, I.M.; Dawson, J.R.; Pietroni, M.C. & Williams, R. (1973).

Transection of the oesophagus for bleeding oesophageal varices. *Br J Surg*, Vol.60,

Infection, coagulation, and variceal bleeding in cirrhosis. *Gut,* Vol.54, №4, (April

Prognosis of hepatic cirrhosis patients with esophageal or gastric variceal hemorrhage: multivariate analysis. *Hepatobiliary Pancreat Dis Int,* Vol.1, №3,

Lata, J.; Juránková, J.; Husová, L.; Senkyrík, M.; Díte, P.; Dastych, M.; Príbramská, V. &

Lee, K.C.; Yang, Y.Y.; Wang, Y.W.; Lee, F.A.; Loong, C.C.; Hou, M.C.; Lin, H.C. & Lee, S.D.

Levin, J. & Bang, F.B. (1968). Clottable protein in Limulus; its localization and kinetics of its

Lin, R.S.; Lee, F.Y.; Lee, S.D.; Tsai, Y.T.; Lin, H.C.; Lu, R.H.; Hsu, W.C.; Huang, C.C.; Wang,

Thalheimer, U.; Triantos, C.K.; Samonakis, D.N.; Patch, D. & Burroughs, A.K. (2005).

Zhao, C.; Chen, S.B.; Zhou, J.P.; Xiao, W.; Fan, H.G.; Wu, X.W.; Feng, G.X. & He, W.X. (2002).

(May 2009), pp. 195-201

pp. 84-87

1110

186-197

*Hepatol,* Vol.18, №4, (April 2003), pp. 424-429

Vol.55, №7, (July 2010), pp. 2077-2085

№8, (August 1973), pp. 646-649

(August 2002), pp. 416-419

2005), pp. 556-563

Liver microcirculation means that the blood circulation is sinusoids as a center, from the end branch of portal vein and hepatic artery through sinusoids to central vein in the liver. Liver microcirculation has two characteristics. One character is there are two import systems from portal vein and hepatic artery. Another is sinusoids with unique structure. All blood vessels which diameters are less 300m are part of microcirculation system. These vessels are able to regulate the blood flow and distribution.1

Sinusoids are a capillary bed which is major place for regulation of blood and exchange of material in the liver. Sinusoids are constituted by microvasculature, sinusoid endothelial cells(SEC), Kupffer cells (KC)and hepatic stellate cells(HSC). 2 SEC and HSC are considered as cellular factors for regulation of sinusoids blood flow. HSC is located in perisinusoids with functions include storing Vitamin A, and controlling the diameter and blood flow of sinusoids by its pseudopods around sinusoids.3 SEC, KC and HSC all have contractility because they contain fiber, microvasculature and contractile protein.2

The liver complex functions, which are biological synthesis, metabolism, detoxify and host defense, are tightly dependent perfect liver microcirculation.4 It is showed that failure of liver microcirculation is a general pathological alteration in chronic hepatitis and liver cirrhosis because changed construction of microvasculature, capillarization of sinusoid (the collagen is filled in perisinusoids) and compression of fibrous tissue. Generally, stenosis of sinusoid will lead to increase of resistence, decrease of blood flow rate and flow volume, and formation of portal hypertension. It is indicated that after hepatitis B virus infection, platelets were recruited to the liver, and their activation correlated with severely reduced sinusoidal microcirculation, delayed virus elimination and increased immunopathological liver cell damage.5

Traditional Chinese Medicine

**2.3 Results** 

1.1

1.0

.9

.8

.7

**2.3.1 In patients with small varices level** 

随访访访(m)

Follow-up (months)

Can Improve Liver Microcirculation and Reduce Portal Hypertension in Liver Cirrhosis 95

level, and moderate and severe level. The randomization code was generated by SAS software for each subject. For patients with small varices, the trial was double blinded. The patients were randomly assigned to 2 groups: Patients were treated with FZHYC in FZHYC group or received placebo in control group. For patients with moderate and severe varices, the trial was single blind. They were divided into 3 groups: FZHYC group in which patients were treated with FZHYC, propranolol group in which patients were treated with Propranolol and combination group in which patients received both FZHYC and propranolol. The dose of FZHYC was 15 capsules (0.3g per capsule) per day and orally administrated at 3 times. The placebo was composed of stir-fried wheat powder with an identical-appearance in the aspects of dosage, color of the contents and packaging. The clinical intervention period lasts 2 years and the follow up would stop once the end points

A total of 6 patients reached the primary end point of esophageal variceal bleeding: 1 of 29 patients in the Fuzheng Huayu group and 5 of 27 patients in the control group. The actuarial probability of remaining free of esophageal variceal bleeding was a significant different between 2 groups (96.3% vs. 77.01%, *P*=0.0422) (Fig. 1). Some patients with small varices were willing to be re-examined by endoscopy after treatment and small varices was dispared in several patients. There was a significant difference of esophageal varices degree

occur. The primary end point was esophageal or gastric variceal bleeding.

in FZHYC group compared to the control group (*P*=0.014). (Tab. 1 and Fig. 2).

0 6 12 18 24 30

Fig. 1. Actuarial probability of remaining free of esophageal variceal bleeding in cirrhotic patients with small varices level. The patients were treated with Fuzheng Huayu Capsule in FZHYC group or placebo in control group for 24 months. Actuarial probability of remaining free of esophageal variceal bleeding was higher in FZHYC group than in controls (*P*<0.05).

GROUP

扶扶扶扶扶扶 1.00-censored

FZHYC group Follow-up (months)

Control group Follow-up (months)

.00-censored

安安剂

In normal condition, liver microcirculation is regulated by 3 ways. (1) SEC is a regulator of blood flow in the liver. The liver injury will lead to injury of SEC, reduction of fenestrae in SEC, straitens of sinusoid within accumulation of erythrocytes and formation of microthrombus. Injury and necrosis of SEC are major causes of disorder of liver microcirculation. (2) KC can also regulate the liver blood flow by its pseudopod inner of sinusoid. Actived KC may lead stenosis of sinusoid due to conglutination of leukocytes.6 (3) HSC is able to directly regulate contraction of sinusoids in normal liver. Active HSC has stronger character of contraction with increased concentration of Ca2+. HSC is just located in the site of sinusoid contraction. Propotional relationship has been proved between contraction of HSC and activation of HSC both *in vivo* and *in vitro*.7,8 Many chemical compounds are as inductors to induce contraction of HSC. They are included Endothelin-1 (ET-1), Angiotensin II and vasopressin. ET-1 is concerned a strongest cytokine to induce contraction of HSC. Perfusion of ET-1 through portal vein leads to contraction of sinusoids. However, the compounds of NO, CO and adrenal medullar hormone have the functions to against ET-1-stimulated contraction of HSC. Contraction of active HSC (myofibroblast cell), which locates in fibrous septum in cirrhotic liver, can change construction of hepatic lobule and lead to increase resistens of microcirculation in the liver.

Once failure of microcirculation in the liver, some symptoms and signs of patient are similar "blood stasis" in Traditional Chinese Medicine. Those are twinge in inside right flank or rib, hepatomegaly or splenomegaly, dim complexion, accompanied with spider nevi and liver palms, deep-red tongue, ecchymosis, taut or thready pulse. Some Chinese herb medicine, which is able to promote blood circulation to remove blood stasis, can prevent esophageal variceal bleeding on cirrhotic patients with portal hypertension, improve liver microcirculation and decrease portal hypertension through treating cirrhotic rats *in vivo* and inhibit contraction of active HSC *in vitro* in resent 10 years.9

Fuzheng Huayu Capsule (FZHYC), a traditional Chinese medicine recipe, has demonstrated the effect of anti-fibrosis and reduced portal vein pressure in patients with chronic hepatitis B.10 Fuzheng means supporting the healthy energy. Huayu means dispersing blood stasis. The recipe is composed by 6 herbs those are Dan Shen (*Radix Salviae Miltiorrhizae*), Chong Cao (*Paecilomyces hepiali Chen & Dai*), Tao Ren (*Semen Persicae*), Jiao Gu Lan (*Gynostemma pentaphyllum*), Song Hua Fen (*Pinus armandii Franch*) and Wu Wei Zi (*Fructus Schisandrae Chinensis*). FZHYC has been used in numerous studies in China and has been found to have a satisfactory prophylaxis effect on the chronic liver injury and hepatic fibrosis in rats and humans. In addition, it enhances hepatic fibrolysis.11

#### **2. The clinical research**

#### **2.1 Aims**

To elucidate the role of FZHYC in the prevention of esophageal variceal bleeding on cirrhotic patients with portal hypertension.

#### **2.2 Methods**

In a multi-center randomized and placebo-controlled trial, 146 cirrhotic patients, whose age range was between 18 and 70 years old, with esophageal varices were enrolled. According to the degree of esophageal and gastric varices, patients were stratified as 2 levels: small level, and moderate and severe level. The randomization code was generated by SAS software for each subject. For patients with small varices, the trial was double blinded. The patients were randomly assigned to 2 groups: Patients were treated with FZHYC in FZHYC group or received placebo in control group. For patients with moderate and severe varices, the trial was single blind. They were divided into 3 groups: FZHYC group in which patients were treated with FZHYC, propranolol group in which patients were treated with Propranolol and combination group in which patients received both FZHYC and propranolol. The dose of FZHYC was 15 capsules (0.3g per capsule) per day and orally administrated at 3 times. The placebo was composed of stir-fried wheat powder with an identical-appearance in the aspects of dosage, color of the contents and packaging. The clinical intervention period lasts 2 years and the follow up would stop once the end points occur. The primary end point was esophageal or gastric variceal bleeding.

#### **2.3 Results**

94 Portal Hypertension – Causes and Complications

In normal condition, liver microcirculation is regulated by 3 ways. (1) SEC is a regulator of blood flow in the liver. The liver injury will lead to injury of SEC, reduction of fenestrae in SEC, straitens of sinusoid within accumulation of erythrocytes and formation of microthrombus. Injury and necrosis of SEC are major causes of disorder of liver microcirculation. (2) KC can also regulate the liver blood flow by its pseudopod inner of sinusoid. Actived KC may lead stenosis of sinusoid due to conglutination of leukocytes.6 (3) HSC is able to directly regulate contraction of sinusoids in normal liver. Active HSC has stronger character of contraction with increased concentration of Ca2+. HSC is just located in the site of sinusoid contraction. Propotional relationship has been proved between contraction of HSC and activation of HSC both *in vivo* and *in vitro*.7,8 Many chemical compounds are as inductors to induce contraction of HSC. They are included Endothelin-1 (ET-1), Angiotensin II and vasopressin. ET-1 is concerned a strongest cytokine to induce contraction of HSC. Perfusion of ET-1 through portal vein leads to contraction of sinusoids. However, the compounds of NO, CO and adrenal medullar hormone have the functions to against ET-1-stimulated contraction of HSC. Contraction of active HSC (myofibroblast cell), which locates in fibrous septum in cirrhotic liver, can change construction of hepatic lobule

Once failure of microcirculation in the liver, some symptoms and signs of patient are similar "blood stasis" in Traditional Chinese Medicine. Those are twinge in inside right flank or rib, hepatomegaly or splenomegaly, dim complexion, accompanied with spider nevi and liver palms, deep-red tongue, ecchymosis, taut or thready pulse. Some Chinese herb medicine, which is able to promote blood circulation to remove blood stasis, can prevent esophageal variceal bleeding on cirrhotic patients with portal hypertension, improve liver microcirculation and decrease portal hypertension through treating cirrhotic rats *in vivo* and

Fuzheng Huayu Capsule (FZHYC), a traditional Chinese medicine recipe, has demonstrated the effect of anti-fibrosis and reduced portal vein pressure in patients with chronic hepatitis B.10 Fuzheng means supporting the healthy energy. Huayu means dispersing blood stasis. The recipe is composed by 6 herbs those are Dan Shen (*Radix Salviae Miltiorrhizae*), Chong Cao (*Paecilomyces hepiali Chen & Dai*), Tao Ren (*Semen Persicae*), Jiao Gu Lan (*Gynostemma pentaphyllum*), Song Hua Fen (*Pinus armandii Franch*) and Wu Wei Zi (*Fructus Schisandrae Chinensis*). FZHYC has been used in numerous studies in China and has been found to have a satisfactory prophylaxis effect on the chronic liver injury and hepatic fibrosis in rats and

To elucidate the role of FZHYC in the prevention of esophageal variceal bleeding on

In a multi-center randomized and placebo-controlled trial, 146 cirrhotic patients, whose age range was between 18 and 70 years old, with esophageal varices were enrolled. According to the degree of esophageal and gastric varices, patients were stratified as 2 levels: small

and lead to increase resistens of microcirculation in the liver.

inhibit contraction of active HSC *in vitro* in resent 10 years.9

humans. In addition, it enhances hepatic fibrolysis.11

cirrhotic patients with portal hypertension.

**2. The clinical research** 

**2.1 Aims** 

**2.2 Methods** 

#### **2.3.1 In patients with small varices level**

A total of 6 patients reached the primary end point of esophageal variceal bleeding: 1 of 29 patients in the Fuzheng Huayu group and 5 of 27 patients in the control group. The actuarial probability of remaining free of esophageal variceal bleeding was a significant different between 2 groups (96.3% vs. 77.01%, *P*=0.0422) (Fig. 1). Some patients with small varices were willing to be re-examined by endoscopy after treatment and small varices was dispared in several patients. There was a significant difference of esophageal varices degree in FZHYC group compared to the control group (*P*=0.014). (Tab. 1 and Fig. 2).

随访访访(m) Follow-up (months)

Fig. 1. Actuarial probability of remaining free of esophageal variceal bleeding in cirrhotic patients with small varices level. The patients were treated with Fuzheng Huayu Capsule in FZHYC group or placebo in control group for 24 months. Actuarial probability of remaining free of esophageal variceal bleeding was higher in FZHYC group than in controls (*P*<0.05).

Traditional Chinese Medicine

1.1

1.0

.9

.8

.7

.6

.5

**cirrhotic rats 3.1.1 Aims** 

in cirrhotic rats.

**3.1.2 Methods** 

for same duration.

随访访访(m)

Follow-up (months)

FZHYC group, but the difference was not significant.

**3.1.2.1 Establishment of liver cirrhosis model** 

**3.1.2.2 Grouping animals and drug administration** 

Can Improve Liver Microcirculation and Reduce Portal Hypertension in Liver Cirrhosis 97

GROUP 联 联用 3.00-censored 普普普尔联 2.00-censored 扶扶扶扶扶扶联 1.00-censored

Combination group

Propranolol group

FZHYC group

0 6 12 18 24 30

Fig. 3. Actuarial probability of remaining free of esophageal variceal bleeding in cirrhotic patients with moderate and severe varices level The patients were treated with Fuzheng Huayu Capsule in FZHYC group, Propranolol in Propranolol group, or hoth in combination group for 24 months. Actuarial probability of remaining free of esophageal variceal bleeding was higher in combination group and FZHYC group than in Propranolol group. Compared with Propranolol group, the difference was significant in combination group (*P*<0.01) and in FZHYC group (*P*<0.05). Although the probability was higher in combination group than in

**3.1 Experiment 1: Chinese herb medicine can decrease portal vein pressure in** 

To investigate effect of Fuzheng Huayu Recipe and SA-B on decreasing portal vein pressure

The Sprague-Dawley male rats were randomly divided into model group (n=46) and normal group (n=14).The rats in model group were administrated intraperitoneally with 0.5% dimethylnitrosamine (DMN) at a dose of 2ml/kg body weight for 3 consecutive days and interval 4 days per week. The treatment was for 4 weeks. The rats in normal group were administrated intraperitoneally with physiological salin at a dose of 2ml/kg body weight

After molding, two rats were randomly sacrificed to observe pathological changes of the liver tissue. The rats in model group were randomly more divided into 3 groups, control


Note: compared with control group,\*: *P*=0.014

Table 1. Difference of developing small varices in FZHYC group and in controls after 24 months

#### **pre treatment post treatment**

Fig. 2. Alteration of small varices by endoscopy examination at pre and post treatment of FZHYC. Arrow shows the small varices in a cirrhotic patient at pre treatmtnt (left). The small varices, however, was disappeared in same patient at post treatmtnt (right).

#### **2.3.2 In patients with moderate and severe varices level**

5 patients in FZHYC group (n=30), 8 patients in Propranolol group (n=30) and 3 patients in the combination group (n=30) had esophageal variceal bleeding during their follow-up. Compared to the Propranolol group (56.99%), there were significant differences in the actuarial probability of remaining free of esophageal variceal bleeding in FZHYC group (76.13%, *P*=0.0131) and the combination group (87.55%, *P*=0.0086). Remaining free of variceal bleeding has a higher trend in combination group, but there was no significant difference between combination group and FZHYC group (*P*=0.3876) (Fig.3).

#### **2.4 Summary**

FZHYC could effectively reduce the risk of esophageal variceal bleeding for cirrhotic patients with varices; especially the combination of the capsule and Propranolol delivered a better effect. The capsule could reduce the varices size in patients with small ones.

#### **3. The experiment** *in vivo*

For studying the mechanism of Fuzheng Huayu Capsule on prevention of esophageal variceal bleeding, we performed several experiments. *Radix Salviae Miltiorrhizae* is a major herb medicine in Fuzheng Huayu Recipe. Salvianolic-acid B (SA-B), which was extracted from *Radix Salviae Miltiorrhizae*, has been demonstrated to inhibit proliferation and functions of HSC *in vitro* 12and anti-fibrosis *in vivo.*13 We did lot of work to deal with how Fuzheng Huayu Recipe and SA-B relieves portal hypertension.

Fig. 3. Actuarial probability of remaining free of esophageal variceal bleeding in cirrhotic patients with moderate and severe varices level The patients were treated with Fuzheng Huayu Capsule in FZHYC group, Propranolol in Propranolol group, or hoth in combination group for 24 months. Actuarial probability of remaining free of esophageal variceal bleeding was higher in combination group and FZHYC group than in Propranolol group. Compared with Propranolol group, the difference was significant in combination group (*P*<0.01) and in FZHYC group (*P*<0.05). Although the probability was higher in combination group than in FZHYC group, but the difference was not significant.

#### **3.1 Experiment 1: Chinese herb medicine can decrease portal vein pressure in cirrhotic rats**

#### **3.1.1 Aims**

96 Portal Hypertension – Causes and Complications

Groups n No varices(n) No changes(n) Enlarged varices(n)

Table 1. Difference of developing small varices in FZHYC group and in controls after 24

Fig. 2. Alteration of small varices by endoscopy examination at pre and post treatment of FZHYC. Arrow shows the small varices in a cirrhotic patient at pre treatmtnt (left). The small varices, however, was disappeared in same patient at post treatmtnt (right).

**pre treatment post treatment** 

5 patients in FZHYC group (n=30), 8 patients in Propranolol group (n=30) and 3 patients in the combination group (n=30) had esophageal variceal bleeding during their follow-up. Compared to the Propranolol group (56.99%), there were significant differences in the actuarial probability of remaining free of esophageal variceal bleeding in FZHYC group (76.13%, *P*=0.0131) and the combination group (87.55%, *P*=0.0086). Remaining free of variceal bleeding has a higher trend in combination group, but there was no significant

FZHYC could effectively reduce the risk of esophageal variceal bleeding for cirrhotic patients with varices; especially the combination of the capsule and Propranolol delivered a

For studying the mechanism of Fuzheng Huayu Capsule on prevention of esophageal variceal bleeding, we performed several experiments. *Radix Salviae Miltiorrhizae* is a major herb medicine in Fuzheng Huayu Recipe. Salvianolic-acid B (SA-B), which was extracted from *Radix Salviae Miltiorrhizae*, has been demonstrated to inhibit proliferation and functions of HSC *in vitro* 12and anti-fibrosis *in vivo.*13 We did lot of work to deal with how Fuzheng

difference between combination group and FZHYC group (*P*=0.3876) (Fig.3).

better effect. The capsule could reduce the varices size in patients with small ones.

**2.3.2 In patients with moderate and severe varices level** 

FZHYC \* 15 8 5 2 control 9 1 3 5

Note: compared with control group,\*: *P*=0.014

months

**2.4 Summary** 

**3. The experiment** *in vivo* 

Huayu Recipe and SA-B relieves portal hypertension.

To investigate effect of Fuzheng Huayu Recipe and SA-B on decreasing portal vein pressure in cirrhotic rats.

#### **3.1.2 Methods**

#### **3.1.2.1 Establishment of liver cirrhosis model**

The Sprague-Dawley male rats were randomly divided into model group (n=46) and normal group (n=14).The rats in model group were administrated intraperitoneally with 0.5% dimethylnitrosamine (DMN) at a dose of 2ml/kg body weight for 3 consecutive days and interval 4 days per week. The treatment was for 4 weeks. The rats in normal group were administrated intraperitoneally with physiological salin at a dose of 2ml/kg body weight for same duration.

#### **3.1.2.2 Grouping animals and drug administration**

After molding, two rats were randomly sacrificed to observe pathological changes of the liver tissue. The rats in model group were randomly more divided into 3 groups, control

Traditional Chinese Medicine

compared with controls. (Fig. 4)

(*P* <0.05). (Fig. 6)

Can Improve Liver Microcirculation and Reduce Portal Hypertension in Liver Cirrhosis 99

large bleeding area, a lot of swell or necrotic hepatocytes, a lot of infiltrating inflammatory cells including lymphocytes and mononuclears, distorted sinusoids and obviously widened portal areas in controls. They were varying decreased that was degeneration or necrosis of hepatocytes, and intrahepatic hemorrhage both in FZHY group and in SA-B group

A little collagen fibers located only in portal areas or surrounded central veins in normal rat liver. Increased fibrous tissue formed thick interval that invaded into the hepatic lobules and moved round to form pseudolobules with different size in controls'liver. The fibrous intervals were thinner both in FZHY group and SA-B group than in controls. It means that

**3.1.3.2.3 The liver function of cirrhotic rats can be improved by Chinese herb medicine**  Compared with normal rats, it was significantly increased in controls that are the activity of serum ALT and AST, serum level of TBiL(*P*<0.01 and *P*<0.05)but meanwhile the concentration of serum Alb were significantly decreased(*P*<0.05). However, they were obviously lower both in FZHY group and SA-B group than in controls that are the activity of serum ALT and AST(*P*<0.01), and serum level of TBiL(*P*<0.05). The concentration of serum Alb were back to normal level with significantly difference compared with controls

Fig. 4. Fuzheng Huayu Recipe and SA-B could relieve inflammation and structure damage in the liver tissue of DMN model rats A: normal group, B: control group, C: FZHY group

and D: SA-B group. The staining was by HE (200).

**3.1.3.2.2 Hepatic fibrosis was reversed by Chinese herb medicine** 

hepatic fibrosis was reversed by FZHY recipe and SA-B. (Fig. 5)

**A B** 

**C D** 

group (n=13), Fuzheng Huayu (FZHY) group (n=15) and the SA-B group (n=16), once liver cirrhosis was confirmed. The rats in the FZHY group were gavaged with extractor of Fuzheng Huayu Recipe. The content of extractor equals 36.9g crude drug per 100ml. The rats in the SA-B group were gavaged with SA-B solution (125mg per 100ml). Simultaneously, the rats in control group and normal group were gavaged with water. All dose of gavage was 1ml/100g body weight, once per day, for 3 weeks.

#### **3.1.2.3 Measurement of portal vein pressure**

The rats were anesthetized with 2% sodium pentobarbital (2ml/kg body weight) in the end of experiment. One PE-10 tube filling with heparin was inserted from superior mesenteric vein into portal vein and then connected with pressotransducer to measure pressure. Enterocoelia was exposed totally after mesurement. If ascites was found, dry cotton ball was used to blot liquid and then it was weighted for measurement of ascites. Serum separated from blood which was taken in the vena cava was used for liver function detection. The sample of liver tissue, which was 1.0cm0.8cm0.3cm, was fixed by formalin, embedded by paraffin, and cutted for 4µm thick for HE and collagen staining. Moreover, 100mg liver tissue was used to detect hydroxyproline (Hyp).

#### **3.1.3 Result**

#### **3.1.3.1 Investigating death and ascites in the rats**

During making hepatic cirrhosis model, there were no dead rats in week 4 but the death started in week 5. Until the end of week 7, there was still no death in normal group but there were 4 dead rats in control group, 2 dead rats both in FZHY group and in SA-B group, respectively. The reason that leads to death was liver failure. The number of the rats with ascites was most in control group than in other groups. The difference was significantly. (Tab. 2)


Note: compared with control group, \*: *P*<0.05

Table 2. The situation of death and ascites in cirrhotic rats in each group

#### **3.1.3.2 Observation of Pathology of liver in rats**

#### **3.1.3.2.1 Inflammation was reduced by Chinese herb medicine in the liver of cirrhotic rats**

The structure of liver lobule was clear, hepatocytes arranged as cords radially from central vein to the periphery and a few connective tissue was observed in the portal area in normal group. At the end of treatment, pathological alteration was observed in the liver that was large bleeding area, a lot of swell or necrotic hepatocytes, a lot of infiltrating inflammatory cells including lymphocytes and mononuclears, distorted sinusoids and obviously widened portal areas in controls. They were varying decreased that was degeneration or necrosis of hepatocytes, and intrahepatic hemorrhage both in FZHY group and in SA-B group compared with controls. (Fig. 4)

#### **3.1.3.2.2 Hepatic fibrosis was reversed by Chinese herb medicine**

98 Portal Hypertension – Causes and Complications

group (n=13), Fuzheng Huayu (FZHY) group (n=15) and the SA-B group (n=16), once liver cirrhosis was confirmed. The rats in the FZHY group were gavaged with extractor of Fuzheng Huayu Recipe. The content of extractor equals 36.9g crude drug per 100ml. The rats in the SA-B group were gavaged with SA-B solution (125mg per 100ml). Simultaneously, the rats in control group and normal group were gavaged with water. All

The rats were anesthetized with 2% sodium pentobarbital (2ml/kg body weight) in the end of experiment. One PE-10 tube filling with heparin was inserted from superior mesenteric vein into portal vein and then connected with pressotransducer to measure pressure. Enterocoelia was exposed totally after mesurement. If ascites was found, dry cotton ball was used to blot liquid and then it was weighted for measurement of ascites. Serum separated from blood which was taken in the vena cava was used for liver function detection. The sample of liver tissue, which was 1.0cm0.8cm0.3cm, was fixed by formalin, embedded by paraffin, and cutted for 4µm thick for HE and collagen staining. Moreover, 100mg liver

During making hepatic cirrhosis model, there were no dead rats in week 4 but the death started in week 5. Until the end of week 7, there was still no death in normal group but there were 4 dead rats in control group, 2 dead rats both in FZHY group and in SA-B group, respectively. The reason that leads to death was liver failure. The number of the rats with ascites was most in control group than in other groups. The difference was significantly.

Groups n Death(n) Ascites(n)

2 2

**3.1.3.2.1 Inflammation was reduced by Chinese herb medicine in the liver of cirrhotic rats**  The structure of liver lobule was clear, hepatocytes arranged as cords radially from central vein to the periphery and a few connective tissue was observed in the portal area in normal group. At the end of treatment, pathological alteration was observed in the liver that was

 1\* 0\*

normal 14 0 0 control 13 4 6

Table 2. The situation of death and ascites in cirrhotic rats in each group

dose of gavage was 1ml/100g body weight, once per day, for 3 weeks.

**3.1.2.3 Measurement of portal vein pressure** 

tissue was used to detect hydroxyproline (Hyp).

**3.1.3.1 Investigating death and ascites in the rats** 

15 16

**3.1.3.2 Observation of Pathology of liver in rats** 

Note: compared with control group, \*: *P*<0.05

**3.1.3 Result** 

(Tab. 2)

FZHY SA-B

A little collagen fibers located only in portal areas or surrounded central veins in normal rat liver. Increased fibrous tissue formed thick interval that invaded into the hepatic lobules and moved round to form pseudolobules with different size in controls'liver. The fibrous intervals were thinner both in FZHY group and SA-B group than in controls. It means that hepatic fibrosis was reversed by FZHY recipe and SA-B. (Fig. 5)

#### **3.1.3.2.3 The liver function of cirrhotic rats can be improved by Chinese herb medicine**

Compared with normal rats, it was significantly increased in controls that are the activity of serum ALT and AST, serum level of TBiL(*P*<0.01 and *P*<0.05)but meanwhile the concentration of serum Alb were significantly decreased(*P*<0.05). However, they were obviously lower both in FZHY group and SA-B group than in controls that are the activity of serum ALT and AST(*P*<0.01), and serum level of TBiL(*P*<0.05). The concentration of serum Alb were back to normal level with significantly difference compared with controls (*P* <0.05). (Fig. 6)

Fig. 4. Fuzheng Huayu Recipe and SA-B could relieve inflammation and structure damage in the liver tissue of DMN model rats A: normal group, B: control group, C: FZHY group and D: SA-B group. The staining was by HE (200).

Traditional Chinese Medicine

Alb (g/L)

**Chinese herb medicine** 

0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1

TBil(g/L)

Can Improve Liver Microcirculation and Reduce Portal Hypertension in Liver Cirrhosis 101

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normal controls FZHY SA-B
