Dissertation
Dissertation > Medicine, health > Chinese Medicine > Of Pharmacy > Pharmacology > Chinese medicine Experimental Pharmacology

The Effect of Traditional Chinese Medcine Huazhuo Jiedu Tiaogan on the Expression of TGF-β1 and Smad3 of Experimental Hepatic Fibrosis in Rats

Author LouYingYing
Tutor LiDianGui
School Hebei Medical University
Course Traditional Chinese Medicine
Keywords ZhuoDu theory Huazhuo Jiedu Tiaogan’formula hepatic fibrosis transforming growth factor-beta1-1 Smad3 Channel protein
CLC R285.5
Type Master's thesis
Year 2010
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Objectives:We used the pig’s serum to set the hepatic fibrosis rats’model by intraperitoneal injection.we observed the effect of Huazhuo Jiedu Tiaogan’formula on the levels of serum HA,PCⅢand Pathomorphology in hepatic fibrosis rats,the expression of TGF-β1 and protein smad3 in liver tissue of hepatic fibrosis rats.Investigated on Pathological Mechanism of Hepatic fibrosis,so that provided theoretical and experimental basis for treating and preventing liver fibrosis.Methods:Total 50 clean male Wistar rats weighed 200~220 grams were randomly divided into five groups:the normal group,the model group, colchicine treatment group,Huazhuo Jiedu Tiaogan’formula low dose group(HZD),Huazhuo Jiedu Tiaogan’formula high dose group(HZG) with 10 rats for each group.Each group was freely fed with ordinary particles,water in the animal laboratory(temperature:22±2℃,humidity:70%±5%).In addition to the normal group,pig’s serum(0.5ml per mouse,)was established in rats by intraperitoneal injection for eight weeks.The time was fixed at 9am on Monday and Friday morning.The drugs were given through stomach-perfusion to HZD group(1.5g/ml),HZG group(3.0g/ml) and colchicine treatment group(0.154g/kg·d) after the model was successful.The rats in the normal control and model groups received intragastric administration of 0.9% saline(10ml/kg).The rats were killed in each group at the 12th weekend.We draw blood from abdominal aorta.The level of HA and PCⅢwas measured with radioimmunoassay(RIA).The specimens were taken and the degree of hepatic fibrosis was judged by routine haematoxylin-eosin staining.The expression of TGF-β1 protein was determined using immunohistochemical technique.The expression of Smad3 was mensurated by Western- immunoblotting method.Data was collected and analyzed through t test or analysis of variance. The statistic software was SPSS13.0.Results1 The ecumenic states of ratsthere was 1 rat dead in model group and the rats in normal control group were all survival with good appetite,smooth hair,good mental state and gradual weight gain.The rats in model group were apathetic, anorexia.The hair were pell-mell and the urine was yellow.There was blood in Some rats’noses.The weight was not increased obviously compared with the normal control group.The rats of the other three treatment group were in good condition with gradual weight gain.But there was no significant difference compared with the normal control group.2 Effect of Huazhuo Jiedu Tiaogan’formula to pathomorphology in hepatic fibrosis ratsWith the macroscopic observation,The liver of normal group was soft,fresh,and smooth with no stain and prominency.The colour was deep red.The liver of model group was hard,big and rough with prominency.The brink was blunt.The colour was dark red.The liver of western medicine and TCM treatment group was not swollen obviously.The liver was crisp,smooth with a slight dark red colour.Sometimes there was hyperplasia as silver sand.Under the light microscope,the hepatic lobules of normal group were arranged in order and ranged radially around the central vein.There was not degeneration and necrosis in liver tissue.The fibrous tissue was expressed little around the central vein.The Structure of model control rats was destroyed. Most of hepatocytes were hydropic degeneration ,ballooning degeneration and necrosis.There were neutrophilic granulocytes and lymphocytes around central vein and portal area.The fibrous tissue was in proliferation.Pseudolobule proliferation could be seen in individual animal.The degree of hepatocyte injury in all medicine intervention group was less serious than model group.But the pathomorphology between the colchicine treatment group and TCM treatment group had no significant difference.3 Effect of Huazhuo Jiedu Tiaogan’formula on the levels of serum HA,PCⅢin hepatic fibrosis ratsCompared with the normal control group,the levels of serum HA,PCⅢin model group rats could be increased with significant difference(P<0.01). Compared with the model control group,The levels of serum HA,PCⅢcould be decreased from all medicine intervention group significantly at the end of 12weeks(P<0.01).The difference between the colchicine treatment group and HZD group was not significant(P>0.05).The level of serum HA,PCⅢin HZG group was significantly reduced compared with colchicine treatment group (P<0.05).4 Effect of Huazhuo Jiedu Tiaogan’formula on TGF-β1 protein of the hepatic fibrosis ratsThe positive expression cells of normal group was a few. It could be manifested as heterogeneously claybank particles.Compared with the normal control group,the cell proportion of TGF-β1 cell expression in the liver tissue of rats from model group were significantly higer than that of the normal group(P<0.05).Compared with the model control group,the level of TGF-β1 protein could be decreased from all medicine intervention group significantly at the end of 12weeks(P<0.05).The difference of TGF-β1 protein between the colchicine treatment group and HZD group was not significant(P>0.05).The expression of TGF-β1 protein in HZG group was significantly reduced compared with colchicine treatment group(P<0.05).5 Effect of Huazhuo Jiedu Tiaogan’formula on Smad3 protein of the hepatic fibrosis ratsCompared with the normal control group,the expression of smad3 in the liver tissue of rats form model group were significantly higer than that of the normal group(P<0.01).Compared with the model control group,the level of Smad3 protein could be decreased from all medicine intervention group significantly(P<0.01).The expression of Smad3 between the colchicine treatment group and HZD group was not significant(P>0.05). The expression of Smad3 in HZG group was significantly reduced compared with the colchicine treatment group(P<0.01).Conclusions1 Huazhuo Jiedu Tiaogan’formula could improve the rats’general condition, and ameliorate hepatocyte injury in hepatic fibrosis rats.It could also reduce the creation of the collagenous fiber and improve the liver pathologic injury.2 Huazhuo Jiedu Tiaogan’formula could significantly reduce serum HA,PCⅢin liver of hepatic fibrosis rats,decrease the deposition of extracellular matrix,exert an anti-hepatic fibrosis effect.3 Huazhuo Jiedu Tiaogan’formula could significantly reduce the expression of TGF-β1 and Smad3 protein.It could inhibit the TGF-β/Smad signal transduction pathway,decrease the deposition of extracellular matrix,and prolong the process of hepatic fibrosis.Therefore,it is practical and feasible to treat hepatic fibrosis on the base of ZhuoDu theory.

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