Study On The Effects Of Supplemental Guizhi Fuling Pill On TNF-? And IL-1 In Liver Organization Of Immune Hepatic Fibrosis Rats

Objective:The objective of this study was to establish a rat model of immune hepatic fibrosis,observe the expression of Supplemental Guizhi Fuling Pill on TNF-?and IL-1 in liver organization of immune hepatic fibrosis rats,and analyze the effect of Supplemental Guizhi Fuling Pill on the prevention and cure of hepatic fibrosis,thus further clarify the mechanism of treating and preventing liver fibrosis with Supplemental Guizhi Fuling Pill,and finally provide a scientific basis for the clinical application and promotion of Supplemental Guizhi Fuling Pill.Methods:Sixty SPF Wistar male rats,aged 6-7 weeks,weighted 180-200 g,were randomly divided into six groups,including negative control group,model group,Daihuang Zhechong Pill control group,high-medium-and low-dose Supplemental Guizhi Fuling Pill treatment groups,with 10 rats.in each group.In model group,0.5ml porcine serum was injected into abdominal cavity,twice per week,incessantly for ten weeks.The rest five groups,except for normal control group which was through intraperitoneal injection of normal saline,were given the same as model group gastric perfusion with Daihuang Zhechong Pill,low-dose Supplemental Guizhi Fuling Pill,medium-dose Supplemental Guizhi Fuling Pill,high-dose Supplemental Guizhi Fuling Pill,respectively once per day.For model and normal control groups,they were treated with distilled water 12.6ml/Kg by gavage.Adjust the amount of dose base on weight of rats each week.After 10 weeks,stop modeling.Weigh rats after fasting 24 hours,then fix in supine position after anesthetization with 2% pentobarbital.Open abdominal cavity to weigh liver and spleen,and obtain 1.5cm*1cm*0.5cm size or so liver tissue with the same position in the right lobe of liver.Then fixe the liver tissue by 10%formaldehyde solution,dehydrate by automatic dehydrator,and embed sections by paraffin.By means of HE staining,observe liver pathomorphology change of rats in each group.Adopt immune histochemistry S-P method to observe the positive expression of TNF-?and IL-1 in rat liver tissue in each group.Results:1.Supplemental Guizhi Fuling Pill can significantly reduce the expression of liver tissue TNF-? in hepatic fibrosis rats.Supplemental Guizhi Fuling Pill loe-dose group showed inflammatory cell infiltration and liver cell edema,also,the number of inflammatory cells and the expression of interstitial cell cytoplasm and periportal was decreased compared with the model group.Supplemental Guizhi Fuling Pill medium-dose group showed interstitial cells,and inflammatory cells expression were significantly reduced compared with the model group.In Supplemental Guizhi Fuling Pill high-dose group,the hepatic lobule structure was close to the normal liver tissues,and the positive expression was mainly located in the cytoplasm of inflammatory cell.The differences between model group and the rest groups were statistically significant(P < 0.01);Dahuang Zhechong Pill control group was higher than that in the Supplemental Guizhi Fuling Pill low-dose,medium-dose and high-dose group(P < 0.01);The difference between blank control group and Supplemental Guizhi Fuling Pill high-dose group was not significant(P > 0.05);The difference between Dahuang Zhechong Pill control group and Supplemental Guizhi Fuling Pilll low-dose group was not significant(P > 0.05).The expression of Supplemental Guizhi Fuling Pill high-dose group is lower than that of the medium-dose group,low-dose group and they were significantly different(P < 0.01).2.Supplemental Guizhi Fuling Pill can significantly reduce the expression of liver hepatic tissue IL-1 in hepatic fibrosis rats.Supplemental Guizhi Fuling Pill low-dose group showed liver lobule structure damage and liver cell degeneration,portal,also,the ex[ression of portal,fibrous septa and around sinusoidal wall was reduced compared with the model group.In Supplemental Guizhi Fuling Pill medium-dose group,the liver lobules injury was relieved,showing visible within fibrous septa and periportal expression,which was significantly reduced compared with the model group.In Supplemental Guizhi Fuling Pill high-dose group,the structure of hepatic lobule damage was significantly improved,showing very few portal tracts and septa expression,which was lighter than the rest of the treatment groups.The difference was significant between the model control group and the rest groups and were significantly different(P < 0.01);Dahuang Zhechong Pill control group was higher than that in the Supplemental Guizhi Fuling Pill medium-dose and high-dose group(P < 0.01);The difference between Dahuang Zhechong Pill control group and Supplemental Guizhi Fuling Pill low-dose group was not significant(P > 0.05).The expression level of Supplemental Guizhi Fuling Pill high-dose group expression level is lower than that of the medium dose-group,low-dose group and they were significantly different(P < 0.01).Conclusion: Supplemental Guizhi Fuling Pill can significantly improve the pathomorphology of liver organization of immune hepatic fibrosis rats and reduce the positive expression of TNF-?and IL-1 in rat liver tissue,and there was a dose-effect relationship in this phenomenon.During the experiment,the rats in Supplemental Guizhi Fuling Pill groups had no obvious side effects and adverse reactions,suggesting that it was safe and reliable.The experiment also showed that TNF-?and IL-1 had a close relationship with the progress and stage of hepatic fibrosis.