| Background:Hepatic fibrosis refers to the extracellular matrix of the liver, especially excessive deposition of collagen. It is not an independent disease but a common pathological process of various chronic liver disease. It also is a necessary development stage to cirrhosis and liver cancer. Therefore antifibrosis treatment has great clinical significance for preventing cirrhosis of the liver and liver cancer, slowing the disease process, reducing the suffering and expenditure of patients. Recent studies show that the activation of hepatic stellate cells, liver sinusoidal capillaries and epithelial-mesenchymal transition are involved in the process of hepatic fibrosis. The studies on Epithelial-mesenchymal transition in liver fibrosis are relatively few, so further study of regulatory factors affect epithelial-mesenchymal transition that pathophysiological phenomenon may provide a new approach for the treatment of liver fibrosis. Western medicine has no particularly effective antifibrotic drugs, however there are a lot of experimental and clinical studies of the Chinese medicine in the prevention and treatment of liver fibrosis. The results has shown that the Chinese medicine has better efficacy and advantages. Yiqihuoxue Prescription(Huangqi, Ezhu, Jiuxiangchong, Taoren) derives from Professor Sun Guizhi’s years of clinical experience of treatment of cirrhosis and fibrosis. A large number of clinical observations by my mentor Lv Wenliang and other experts of Guang An Men Hospital hepatitis outpatient verify that Yiqihuoxue Prescription (YQHXP)does have a good anti-fibrotic efficacy. Our study verify the antifibrotic efficacy of YQHXP from animal experiments and research its mechanism from the point of view of epithelial-mesenchymal transition in order to provide a scientific basis for its better application in clinical.Objective:By observing YQHXP in rats of liver fibrosis, we verify its antifibrotic effect and try to confirm the presence of pithelial mesenchymal transition (EMT) in process of liver fibrosis. Furthermore, we also attempt to prove that the signal pathway of transforming growth factor-β(TGF-β)/transforming growth factor-beta type Ⅱ receptor (TGFβRⅡ)/Smads is involved in the EMT. Finally, we try to explore the molecular mechanism of YQHXP anti-fibrosis in order to increased clinical basis for better used in clinical practice and provide new research ideas for mining the scientific connotation of famous TCM doctor’s experience. Methods:We copied liver fibrosis model of rat using carbon and ethanol. From the fifth week onwards, rats were given drug intervention while modeling.The experiment finished after12weeks of drug intervention. We took the serum, livers and spleens of rats at the end of the experiment. Serum was used for biochemical detection. We weighed livers and spleens, afterwards calculated liver index and spleen index. The same part of each rat liver was taken to HE staining and then observed pathological results in each group under light microscope, finally fibrosis stage was determined. E-cadherin, TGF-betal, TGFβRⅡ and Smad2of liver tissue were detected by immunohistochemical method, and then we applied the Axio Vision Release4.8.2image analysis software to measure the total expression area and percentage of positive area of various indicators.Results:After modeling of carbon tetrachloride and ethanol, liver wet weight, spleen wet weight, liver index and spleen index of the model group are higher than the blank control group, furthermore the difference is statistically significant (P<0.05). After treatment of YQHX these four indicators decrease compared with the model group, and the difference is statistically significant (P<0.05). ALT and AST of model group increase with ALB reduce, in addition the difference between the model group and the blank control group is statistically significant (P<0.05). After treatment of YQHX, ALT and AST level reduce with ALB increase compared with the model group, and the difference is statistically significant (P<0.05).The HE staining show that liver fibrosis of blank control group are all in S0, but model group are mainly concentrated in the S4stage, furthermore the difference between two groups is statistically significant (P<0.01). YQHX group are mainly distributed in the S1to S3period, and the difference is statistically significant (P<0.01) compared with the model group.Immunohistochemical results show that E-cadherin downregulation of liver tissue of the model group with upregulation of TGF-betal, TGFβRⅡ and Smad2. Furthmore the difference between the model group and the blank control group is statistically significant (P<0.05). After treatment of YQHX, E-cadherin upregulation with downregulation of TGF-betal, TGFβRⅡ and Smad2compared with model group. The difference between two groups is statistically significant (P<0.05).Conclusions:YQHXP can reduce liver wet weight, spleen wet weight, liver index and spleen index of rat of liver fibrosis. It also can reduce ALT and AST on the one hand, on the other hand improve the ALB level. In short YQHXP could ameliorate liver and spleen morphology of rat of liver fibrosis while ameliorate liver function. From the pathological point of view, YQHXP has good anti-fibrotic efficacy by ameliorating the degree of inflammation of the liver, reducing the deposition of extracellular matrix and reversing hepatic fibrosis. EMT is involved in the development process of rat liver fibrosis, and decreased expression of E-cadherin of the model group is the evidence of its participation in fibrosis. YQHXP could increase the expression of E-cadherin suggesting that it can reverse the EMT and then reverse liver fibrosis. YQHXP is able to block TGF-β/TGFβRⅡ/Smad pathway by downward adjusting the expression of TGF-β1, TGFβRⅡ and Smad2, thereby block the process of liver fibrosis. In view of the TGF-β/TGFβR Ⅱ/Smads pathway that is one of the main path of the EMT, and our research found YQHXP can not only reverse the EMT but also block TGF-β/TGFβRⅡ/Smads pathway, then we can boldly inferred that YQHXP has good antifibrotic effect by blocking TGF-β/TGFβRⅡ/Smads pathway so as to reverse EMT. |
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