运动联合奥利司他对非酒精性脂肪肝炎的药效作用研究

Study on the pharmacodynamic effects of exercise in combination with orlistat in non-alcoholic steatohepatitis

关键词:  非酒精性脂肪肝炎  运动  奥利司他  脂肪变性  炎症

Key words:  nonalcoholic steatohepatitis  exercise  orlistat  fatty degeneration  inflammation

摘要:

目的：探讨运动联合奥利司他对高脂饮食诱导的小鼠非酒精性脂肪肝炎(NASH)的治疗作用。方法：40只雄性C57BL/6J小鼠按照体重随机分为正常组、模型组、运动组、给药组和联合治疗组，每组8只。每周三次称量并记录小鼠体重，在最后一次给药结束后禁食12小时，测量空腹血糖后全部安乐死，测量各组小鼠体长、肝重、附睾脂肪重量，检测血清胰岛素水平、血清肝功能指标AST、ALT、TC、TG、HDL、LDL和血清炎症因子TNF-α、IL-6、IL-1β表达水平，计算Lee’s指数和胰岛素抵抗指数；HE染色和油红O染色观察肝组织和肝细胞病理变化、脂质蓄积情况和炎症情况；蛋白免疫印迹法(Western blot)和免疫荧光染色(Immunofluorescence, IF)检测肝脏脂质代谢相关蛋白的表达情况。结果：与模型组相比，运动组、给药组和联合治疗组小鼠的体重、肝重、肥胖指数、附睾脂肪重量、肝功能指标AST、ALT、TC、TG、LDL、HDL水平、炎症因子TNF-α、IL-6、IL-1β水平、空腹血糖、胰岛素水平和胰岛素抵抗指数均明显降低；肝脏脂肪变性程度、炎症、NAS评分、脂滴沉积情况均有明显改善；肝脏脂质合成蛋白FASN、ACC水平显著降低，脂质氧化蛋白PPARα、CPT1α水平显著升高，各项指标均以联合治疗组效果最好。结论：运动联合奥利司他对NASH小鼠具有良好的治疗作用，可以显著改善肝功能和胰岛素抵抗，抑制肝脏脂肪沉积和炎症，与激活AMPK相关通路，降低脂质合成蛋白FASN、ACC水平、提高脂质氧化蛋白PPARα、CPT1α含量有关。

Abstract:

Objective: To investigate the therapeutic effect of exercise in combination with orlistat on high-fat diet-induced non-alcoholic steatohepatitis (NASH) in mice. Methods: 40 male C57BL/6J mice were randomly assigned to one of five treatment groups based on their body weight: normal, model, exercise, drug administration, and combined treatment, with 8 mice in each group. The normal group was fed a normal diet, while the other groups were fed a high-fat diet. Treatment began at week 13, with mice in the exercise group receiving 1 hour of exercise 5 times per week, mice in the dosing group receiving orlistat once daily at a dose of 60 mg/kg by gavage, and mice in the combined treatment group receiving both exercise and orlistat at the same time. The dosing cycle lasted 8 weeks, from week 13 to week 20. The body weight of the mice was weighed and recorded three times a week. At the end of the last administration, all mice were fasted for 12 hours and euthanized after measurement of fasting blood glucose levels. The body length, liver weight, and epididymal fat weight of the mice in each group were measured and serum insulin levels, serum liver function indices AST, ALT, TC, TG, HDL, LDL, and the expression levels of inflammatory cytokines TNF-α, IL-6, and IL-1β in serum were determined. Lee's index and insulin resistance index were calculated. HE staining and oil red O staining were used to examine the pathological changes, lipid accumulation, and inflammation of liver tissue and hepatocytes. Western blot and Immunofluorescence were used to detect the expression of FASN, ACC, CPT1α, PPARα, and other proteins in liver lipid metabolism. Results: Compared to the normal group, body weight, liver weight, obesity index, epididymal fat weight, liver function indices AST, ALT, TC, TG, LDL, HDL levels, levels of the inflammatory cytokines TNF-α, IL-6, IL-1β, fasting blood glucose, insulin levels and insulin resistance index were significantly increased. The degree of hepatic steatosis, inflammation, NAS score and lipid droplet deposition were significantly increased. The levels of lipid synthesis proteins FASN and ACC were significantly increased, and the levels of lipid oxidizing proteins PPARα and CPT1α were significantly decreased. Compared to the model group, the weight, liver weight, obesity index, epididymal fat weight, liver function indexes AST, ALT, TC, TG, LDL, HDL, levels of inflammatory cytokines TNF-α, IL-6, IL-1β, fasting blood glucose, insulin level and the insulin resistance index decreased significantly in the exercise group, administration group and combined treatment group. The degree of hepatic steatosis, inflammation, NAS score, and lipid droplet deposition was significantly improved. The levels of lipid synthesis protein FASN and ACC were significantly decreased, and the levels of lipid oxidation proteins PPARα and CPT1α were significantly increased. Conclusion: Exercise in combination with orlistat has a good therapeutic effect in NASH mice. It can significantly improve liver function and insulin resistance, inhibit fat deposition in the liver and inflammation, and is associated with the activation of AMPK-related signaling pathways, the reduction of the levels of lipid synthesis proteins FASN and ACC, and the increase of the levels of lipid oxidation proteins PPARα and CPT1α.