芪参益气滴丸对高脂饮食联合链脲霉素诱导糖尿病小鼠肝损伤的作用及机制

Effect and mechanism of Qishen Yiqi Droplet on liver injury induced by high-fat diet combined with streptozotocin in diabetes mice

关键词:  芪参益气滴丸  糖尿病  肝损伤  肝细胞凋亡  脂肪变性

Key words:  Qishen Yiqi Droplet  diabetes  liver injury  hepatocyte apoptosis  steatosis

摘要:

[目的] 探讨芪参益气滴丸对糖尿病小鼠肝损伤的保护作用及机制。[方法] 80只雄性小鼠随机分为对照组，模型组，芪参益气滴丸低剂量组（227.5 mg/kg）、高剂量组（455 mg/kg）和EX-527抑制剂组。对照组喂养正常饲料，其余4组予高脂饲料，第3周开始连续7 d腹腔注射50 mg/kg链脲霉素，建立2型糖尿病肝损伤模型，对照组注射等剂量生理盐水。第5周予药物干预14 d后，取血清检测天门冬氨酸氨基转移酶（AST）、丙氨酸氨基转移酶（ALT）、总胆固醇（TC）和三酰甘油（TG）水平。病理染色检测肝脏形态结构、胶原纤维面积、肝脂肪变性程度和肝细胞凋亡数量。蛋白免疫印迹法（Western Blot）检测肝组织沉默信息调节因子1（Sirt1）、AMP依赖的蛋白激酶（AMPK）、过氧化物酶体增殖物激活受体γ-共激活因子1-α（PGC-1α）蛋白表达。[结果] 与对照组比较，模型组小鼠血清中AST、ALT、TC和TG均明显升高（P<0.01），肝小叶结构紊乱，细胞核深染、固缩，局部炎性浸润，肝细胞内存在大量红色脂肪滴，肝纤维化面积和肝细胞凋亡数目显著增加（P<0.01），p-AMPKα/AMPKα表达降低（P<0.01），Sirt1和PGC-1α含量显著升高（P<0.05或P<0.01）。芪参益气滴丸低、高剂量组均能降低肝功能损伤指标（P<0.05或P<0.01），抑制肝脏结构病变及肝脂肪变性，减轻肝纤维化损伤，降低肝细胞凋亡数目（P<0.01），提升Sirt1和p-AMPKα/AMPKα水平（P<0.05或P<0.01），下调PGC-1α表达（P<0.01）。而EX-527则部分逆转芪参益气滴丸的保护效应。[结论] 芪参益气滴丸可能通过激活Sirt1/AMPKα/PGC-1α信号，抑制糖尿病小鼠肝纤维化，减轻肝脂肪变性和肝细胞凋亡，进而促进糖尿病肝功能修复。

Abstract:

[Objective] To explore the protective effect and mechanism of Qishen Yiqi Droplets on liver injury in diabetes mice. [Methods] Eighty male mice were randomly divided into control group，model group，Qishen Yiqi Droplets low-dose group（227.5 mg/kg），high-dose group（455 mg/kg），EX-527 inhibitor group. The control group was fed with normal diet，and the other four groups were fed with high-fat diet. From the third week，50 mg/kg streptozotocin was intraperitoneally injected for 7 consecutive days to establish the liver injury model of type 2 diabetes. The control group was injected with equal dose of normal saline. In the fifth week，14 days after Qishen Yiqi Droplets intervention，serum was taken to detect the levels of AST，ALT，TC and TG. The morphology and structure of the liver，the area of collagen fibers，the degree of hepatic steatosis and the number of hepatocyte apoptosis were detected by pathological staining. The expressions of Sirt1，AMPK and PGC-1α were measured by western blot. [Results] Compared with the control group，the serum AST，ALT，TC and TG of diabetes mice were significantly increased （P<0.01），the structure of hepatic lobule was disordered，the nucleus was deeply stained，pyknotic，local inflammatory infiltration，there were a lot of red fat droplets in hepatocyte，the area of liver fibrosis and the number of apoptotic hepatocyte were significantly increased（P<0.01）. The p-AMPKα/AMPKα expression reduced （P<0.01），the content of Sirt1 and PGC-1α was increased significantly （P<0.05 or P<0.01）. Both the low and high dose groups of Qishen Yiqi Droplets can diminish liver function damage indicators（P<0.05 or P<0.01），inhibit the pathological changes of liver structure and liver steatosis，alleviate liver fibrosis damage，reduce the number of hepatocyte apoptosis （P<0.01），and improve Sirt1 and p-AMPKα/AMPKα level （P<0.05 or P<0.01），abate PGC-1α expression（P<0.01）. EX-527 partially reversed the protective effect of Qishen Yiqi Droplets. [Conclusion] Qishen Yiqi Droplets may activate Sirt1/AMPKα/PGC-1α signal，inhibit hepatic fibrosis in diabetes mice，reduce liver steatosis and hepatocyte apoptosis，and then promote the repair of liver function in diabetes.