Experimental Study On Regulation Of FXR Of Intrahepatic Cholestasis With The Treatment Of Dispersing Stagnated Liver Qi For Promoting Bile Flow And Related Clnical Study

BackgroundOur country has a high incidence of liver diseases, various liver diseases could cause liver cholestasis, which will induce liver inflammation or fibrosis even cirrhosis, thus the treatment of cholestatic is very important to delaying the process liver disease. Cholestasis mainly refers to the abnormally elevated concentration of bile acids. FXR as a regulator of bile acid metabolism involved in transcriptional and regulation of bile acid metabolism in varies of related membrane transporter, is the hotspot in treatment of cholestatic research. NTCP can entere bile salts into liver cells, BSEP pump bile salts into the bile duct. Intrahepatic cholestasis is called "jaundice" in Traditional Chinese Medicine, TCM have its distinct character-istics and advantages. Dispersing stagnated liver Qi for promoting bile flow is the basic treatment principles for jaundice. FXR and BESP are considered as the same level as an indicator in most Chinese medicine research. Review of the literature, we found most of them are clinical observations while few prospective studies, most of them are small amounts of clinical cases observed while few randomized, double-blind, multi-center controlled studies.ObjectiveEstablish disease and syndrome combined model with dampness-heat syndrome by the composite factors as hot and damp environment and fat dietary and drug, we observe the effect of Yinzhihuang injection on FXR, NTCP, BESP expression, the serum biochemical parameters and pathological changes in liver tissue of damp-heat syndrome rats, in order to clear Yinzhihuang injection specific targets in the treatment of intrahepatic cholestasis. At the same time, we will evaluate the efficacy of Yinzhihuang injection by clinical experiments. MethodsExperiment on rats:1modeling:we simulate Damp-heat treatment factors, place the rats in hot and humid boxes, fed then with a mixture of glucose and fat pig, and then ANIT which could induce liver injury to make rat model.2grouping and intervention:51male SPF level SD rats were randomly divided into7groups: Normal control Group(7), Model Group(7), Yinzhihuang injection Group(7), GW4064Group(7), Guggulsterone Group(7), Yinzhihuang injection and GW4064Group(8), Yinzhihuang injection and Guggulsterone Group(8).Normal control group (N):Every morning1mL/100g0.9%saline gavage.Model group (M):Every morning1mL/100g0.9%saline gavage.Yinzhihuang injection group (Y):ANIT gavage48hours later, Yinzhihuang intraperitoneal injection with1mL/100g/day for7days.GW4064group (G):ANIT gavage48hours later, GW4064with100mg/kg/day for7days.Guggulsterone Group (GS):ANIT gavage48hours later, Guggulsterone with100mg/kg/day for7days.Yinzhihuang+GW4064Group (+G):ANIT gavage48hours later, Yinzhihuang intraperitoneal injection with1mL/100g/day and GW4064with100mg/kg/day for7days.Yinzhihuang+Guggulsterone Group (+GS):ANIT gavage48hours later, Yinzhihuang intraperitoneal injection with1mL/100g/day and Guggulsterone with100mg/kg/day for7days.3indicatrix and methods:3.1Analyze TE、DB、IB、ALT、AST、ALP、γ-GT in rats serum by automatic biochemical analyzer.3.2Observe the morphological changes in liver tissue by microscopy.3.3Western Blot to detect the rat liver FXR, NTCP, BSEP expression.3.4RT-PCR to detect the rat liver FXR, NTCP, BSEP mRNA expression.Clinical research:Clinical research on Yinzhihuang injection treatment of intrahepatic cholestasis:60patients were observed based on the diagnostic criteria of liver and gallbladder damp-heat syndrome and intrahepatic cholestasis from Spleen and Stomach Diseases Branch of First Affiliated Hospital of Guangzhou University of TCM. Patients were randomly divided into two groups by random number table, Control group:glutathione1.2g/d, SAM1g/d, and UDCA250mg/time, twice/d, Treatment group:Yinzhihuang injection20ml/d on the basis of the control group.Observer TCM symptoms, liver function as well as comparative efficacy of TCM and Western Medicine, evaluate the efficacy of Yinzhihuang Injection for intrahepatic cholestasis.Result1Yinzhihuang injection can improve the symptoms of damp-heat syndrome, reduce TB, DB, ALP, γ-GT and the hepatic pathological damage compared with model group.2Yinzhihuang injection can promote the expression of FXR and the mRNA expression of BESP, there was no significant difference in protein and gene expression of NTCP between Yinzhihuang group and Model group. The expression of NTCP is lower, the higher expression of FXR, BESP, the higher expression of NTCP, lower expression of FXR, BESP. BESP was significantly associated with FXR while NECP has no significant correlation with FXR.3Yinzhihuang group has a significant effect in reducing serum bilirubin and liver enzymes compared with Control group, but there was no significant difference. Yinzhihuang injection combined with western medicine showed better efficacy than the control group, and both of them have no obvious side effects.Conclusion1The model eatablished by hot and damp environment and fat dietary and drug is consistent with the theory of TCM and Western Medicine. Yinzhihuang injection can effectively improve the damp-heat syndrome, reduce TB, DB, ALP, γ-GT and the hepatic pathological damage compared with model group.2Yinzhihuang injection can intervene FXR to promote the expression of BESP for intrahepatic cholestasis.3Yinzhihuang injection combined with western drugs is superior in terms of improving syndromes than medicine group, the effect of lowering serum bilirubin and liver enzymes is significant but there was no significant difference between the two groups. None has toxic side effects.