茵陈蒿汤对MKR鼠2型糖尿病合并非酒精性脂肪性肝病的作用

Effect of Yinchenhao Tang on Type 2 Diabetes Mellitus Complicated with Non-alcoholic Fatty Liver Disease in MKR Mice

目的:探讨茵陈蒿汤对MKR鼠2型糖尿病合并非酒精性脂肪性肝病的作用机制。方法:8周龄MKR小鼠40只高脂喂养8周后,随机分为模型组、茵陈蒿汤原方剂量组(17.16 g·kg^(-1)),茵陈蒿汤教材剂量组(4.68 g·kg^(-1)),二甲双胍+辛伐他汀组[(65+2.6)×10^(-3) g·kg^(-1)],同龄MKR鼠10只作为空白组,FVB鼠10只分别作为正常组。各组灌胃给药8周后,测定小鼠肝脏湿重,口服糖耐量(OGTT),血清炎性因子白细胞介素-6(IL-6),肿瘤坏死因子-α(TNF-α),血脂和肝功能的变化,透射电镜和苏木素-伊红(HE)染色对肝脏组织形态进行观察,蛋白免疫印迹法(Western blot)检测肝脏组织Toll样受体4(TLR4),髓样分化因子88(MyD88),核转录因子-κB(NF-κB)蛋白表达变化。结果:与模型组比较,给药组小鼠肝湿重指数显著降低(P<0.01),OGTT较模型组明显有所改善(P<0.05),血清中IL-6,TNF-α含量显著降低(P<0.01),形态学改变上,茵陈蒿汤对肝细胞结构有保护作用,减少肝细胞脂肪样变性;茵陈蒿汤可以明显下调T2DM合并NAFLD模型小鼠肝脏组织TLR4,MyD88,NF-κB蛋白含量(P<0.05),且对于TLR4,NF-κB的下调,茵陈蒿汤原方剂量明显优于茵陈蒿汤剂量(P<0.05)。结论:茵陈蒿汤能降低炎症因子水平,肝组织TLR4,MyD88,NF-κB的表达水平,改善肝脏病理损伤,干预MKR鼠T2DM合并NAFLD疾病状态,具有一定保护肝功能,降低血脂以及延缓肝脏炎症反应的作用。

Objective:To investigate the action mechanism of Yinchenhao Tang against type 2 diabetes mellitus(T2DM)complicated with non-alcoholic fatty liver disease(NAFLD)in MKR mice.Method:Forty eight-week-old MKR mice were fed a high-fat diet for eight weeks and then divided into the model group,original Yinchenhao Tang(17.16 g·kg^(-1))group,Yinchenhao Tang group at a specified dose(4.68 g·kg^(-1))in teaching materials,and positive drug[metformin+simvastatin,(65+2.6)×10^(-3) g·kg^(-1)]group.Another 10 MKR mice of the same age were classified into the blank group and 10 FVB mice into the normal group.After eight weeks of intragastric administration in each group,the liver wet weight,oral glucose tolerance test(OGTT),serum inflammatory factors interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α),and changes in blood lipid and liver function were determined.Hematoxylin-eosin(HE)staining was conducted for observing the morphological changes in liver tissue under a transmission electron microscope,followed by the detection of Tolllike receptor 4(TLR4),myeloid differentiation factor 88(MyD88),and nuclear transcription factor-κB(NF-κB)protein expression by Western blot.Result:Compared with the model group,the medication groups exhibited significantly reduced liver wet weight index(P<0.01),improved OGTT result(P<0.05),and downregulated serum IL-6 and TNF-αlevels(P<0.01).In terms of morphological changes,Yinchenhao Tang protected the hepatocyte structure and alleviated hepatocyte steatosis.Moreover,Yinchenhao Tang obviously down-regulated the protein expression levels of TLR4,MyD88,and NF-κB in liver tissue of MKR mice with T2DM combined with NAFLD(P<0.05),and the down-regulation of TLR4 and NF-κB in the original Yinchenhao Tang group was better than that in the Yinchenhao Tang group at a specified dose in teaching materials(P<0.05).Conclusion:Yinchenhao Tang is able to reduce inflammatory factor levels and downregulate TLR4,MyD88,and NF-κB expression in liver tissue to relieve the pathological liver injury and interfere with T2DM combined with NAFLD of MKR mice.It exerts a certain liver-protective effect by lowering the blood lipids and delaying the hepatic inflammation.