加味茵陈蒿汤对大鼠胆汁淤积性肝病的治疗作用

目的:探讨加味茵陈蒿汤对α-异硫氰酸萘酯(ANIT)诱导胆汁淤积性肝病(CSLD)大鼠模型的治疗作用。方法:将56只大鼠随机分为7组,分别为空白组,模型组,复方甘草酸苷胶囊组(22.5,45 mg·kg^-1),加味茵陈蒿汤低、中、高剂量组(4.1,8.1,16.2 g·kg^-1)。以ANIT(100 mg·kg^-1)灌胃1次,制备胆汁淤积性肝损伤模型。复方甘草酸苷胶囊及加味茵陈蒿汤在造模第2天开始灌胃给药,连续4 d。第5天进行胆管插管手术测量大鼠胆汁流速,取血清检测各组总胆红素(TBIL),直接胆红素(DBIL),间接胆红素(IBIL),丙氨酸氨基转移酶(ALT)和总胆汁酸(TBA)等血清学指标。采用苏木素-伊红(HE)染色观察肝组织病理变化。采用蛋白免疫印迹法(Western blot)检测大鼠肝组织中G蛋白偶联胆汁酸受体(TGR5),核苷酸结合寡聚化结构域样受体3(NLRP3),半胱氨酸蛋白酶-1(Caspase-1)蛋白表达情况。结果:与空白组比较,模型组胆汁流速显著降低(P<0.01),各血清学指标均显著增高(P<0.01),肝组织病变严重,且肝组织中TGR5,Caspase-1蛋白表达显著增强(P<0.01);与模型组比较,复方甘草酸苷胶囊组对于胆汁流速和血清学指标无显著影响,加味茵陈蒿汤高剂量组胆汁流速增快(P<0.05),各血清学指标均降低(P<0.05),复方甘草酸苷胶囊组和加味茵陈蒿汤组大鼠肝组织病变均有不同程度地改善,且肝组织中TGR5,Caspase-1蛋白表达均明显降低(P<0.05)。结论:加味茵陈蒿汤可有效治疗大鼠CSLD,其机制可能与胆汁酸分子及胆汁酸受体TGR5介导的炎症因子有关。

Therapeutic Effect of Modified Yinchenhao Tang on Cholestatic Liver Disease in Rats

α-isothiocyanate(ANIT)-induced cholestatic liver disease(CSLD).Method:Wistar rats were randomly divided into 7 groups:blank control group,model control group,compound Glycyrrhizin capsules group(22.5,45 mg·kg^-1),modified Yinchenhao Tang low,middle and high dose groups(4.1,8.1,16.2 g·kg^-1).A model of cholestatic liver injury was prepared by intragastric administration of ANIT(100 mg·kg^-1).Glycyrrhizin capsules and modified Yinchenhao Tang were administered intragastrically on the second day of modeling for 4 consecutive days.And bile duct intubation was performed on the fifth day to measure the bile flow rate of the rats,and serum was taken to test the total bilirubin(TBIL),direct bilirubin(DBIL),indirect bilirubin(IBIL),alanine aminotransferase(ALT)and total bile acid(TBA)serological indicators of each group.Pathological changes of liver tissues were observed by hematoxylin-eosin(HE)staining.The expression levels of G proteincoupled bile acid receptor(TGR5),nucleotide binding oligomerization domain-like receptor 3(NLRP3)and cysteinyl aspartate specific proteinase-1(Caspase-1)proteins in the iver tissues were detected by Western blot.Result:Compared with the blank control group,bile flow rate in the model group decreased significantly(P<0.01).TBIL,DBIL,IBIL,ALT and TBA level in serum were significantly increased(P<0.01),liver tissue lesions were severe,and significantly increased the expression of liver tissue TGR5 and Caspase-1.Compare with model group,the compound Glycyrrhizin capsules group had no significant effect on bile flow rate and TBIL,DBIL,IBIL,ALT and TBA level in serum.Bile flow rate increased and TBIL,DBIL,IBIL,ALT and TBA level in serum decreased significantly in modified Yinchenhao Tang high dose group.The compound Glycyrrhizin capsules group and modified Yinchenhao Tang group have different extents of improvement the pathological changes of the lung tissues,and the protein expression of TGR5 and Caspase-1 were significantly decreased in the liver tissue(P<0.01).Conclusion:Modified Yinchenhao Tang can effectively treat CSLD in rats,and its mechanism may be related to bile acid and bile acid receptor TGR5-mediated inflammatory factors.