摘要
目的:探讨左归降糖清肝方改善2型糖尿病(T2DM)合并非酒精性脂肪性肝病骨骼肌特异性胰岛素样生长因子-1受体功能缺失(MKR)小鼠糖脂代谢的作用机制。方法:以MKR小鼠为研究对象,高脂饲料喂养8周诱导形成NAFLD,随机分为模型组、二甲双胍组(0.067 g·kg^(-1))、左归降糖清肝方高、低剂量组(14.8、7.4 g·kg^(-1)),同龄FVB小鼠10只作为正常组。分别药物治疗8周后,予以糖耐量测试(OGTT),取血清检测甘油三酯(TG)、总胆固醇(TC),称取小鼠肝脏湿重,取肝组织进行苏木素-伊红(HE)染色、油红O染色观察组织病理形态变化,实时荧光定量聚合酶链式反应(Real-time PCR)及蛋白免疫印迹法(Western blot)检测肝组织叉头框蛋白O1(FoxO1)、磷酸烯醇式丙酮酸羧激酶(PEPCK)、葡萄糖6磷酸酶(G6Pase)、载脂蛋白C3(ApoC-Ⅲ)的mRNA表达及其蛋白表达情况。结果:与正常组比较,模型组小鼠空腹血糖、肝脏指数、血清TG、血清TC、OGTT水平显著上升(P<0.01);与模型组比较,二甲双胍组及左归降糖清肝方组小鼠空腹血糖、肝脏指数显著下降(P<0.01),左归降糖清肝方高剂量组血清的TG、TC水平均明显下降(P<0.05,P<0.01),二甲双胍组及左归降糖清肝方高剂量组可以降低OGTT水平(P<0.05);在组织病理上,与正常组比较,模型组小鼠可见肝组织脂滴及肝细胞内空泡减少,体积变大;与模型组比较,左归降糖清肝方组和二甲双胍组可见肝组织脂滴减少,肝细胞内空泡减少,体积变小;在分子层面,与正常组比较,模型组小鼠肝组织FoxO1、PEPCK、G6Pase、ApoC-Ⅲ的mRNA水平显著上调(P<0.01),FoxO1、PEPCK、G6Pase、ApoC-Ⅲ的蛋白表达显著上调(P<0.01);与模型组比较,左归降糖清肝方组FoxO1、PEPCK、G6Pase、ApoC-Ⅲ的mRNA水平显著下调(P<0.01),FoxO1、PEPCK、G6Pase、ApoC-Ⅲ的蛋白表达显著下调(P<0.01)。结论:左归降糖清肝方能改善T2DM合并NAFLD糖脂代谢并能修复肝脏病理损伤,可能是通过调控肝组织FoxO1、PEPCK、G6Pase、ApoC-Ⅲ相关蛋白的表达,达到调脂、降糖及延缓肝脏脂肪病变的作用。
Objective:To investigate the mechanism of Zuogui Jiangtang Qinggan prescription(ZJQP)in improving glucose and lipid metabolism in loss of skeletalmuscle-specific insulin-like growth factor-1 receptor function(MKR)mice with type 2 diabetes mellitus(T2DM)and non-alcoholic fatty liver disease(NAFLD).Method:NAFLD was induced by high-fat diet feeding for 8 weeks in MKR mice,which were randomly divided into model group,metformin group(0.067 g·kg^(-1)),and ZJQP high and low-dose groups(14.8,7.4 g·kg^(-1)).Ten FVB mice of the same age were used as the normal group.After 8 weeks of drug treatment,the oral glucose tolerance test(OGTT)was performed,the serum was taken to detect triacylglycerol(TG)and total cholesterol(TC),and the wet weight of the mouse liver was weighed.Haematoxylin-eosin(HE)staining and oil red O staining were performed to assess histopathology of liver.The mRNA expression and protein expression of Fork head box protein O1(FoxO1),phosphoenolpyruvate carboxykinase(PEPCK),glucose-6-phosphatase(G6Pase),and apolipoprotein C3(ApoC-Ⅲ)in liver tissues were detected by real-time fluorescent quantitative PCR(Real-time PCR)and Western blot,respectively.Result:As compared with the normal group,the levels of fasting blood glucose,liver index,serum TG,TC,and OGTT of mice in the model group increased significantly(P<0.01).As compared with model group,the fasting blood glucose and liver index of the mice in the metformin group and the ZJQP group decreased significantly(P<0.01),the serum levels of TG and TC in the high-dose ZJQP group decreased significantly(P<0.05,P<0.01),and the OGTT of mice in the metformin group and the high-dose ZJQP group improved(P<0.05).In histopathology,as compared with the normal group,mice in the model group showed decreased lipid droplets and vacuoles in hepatocytes,and their volumes became larger.Compared with the model group,the ZJQP group and metformin group showed that the lipid droplets in liver tissues were reduced,the vacuoles in liver cells were reduced,and the volume was smaller.At the molecular level,as compared with the normal group,the mRNA and protein levels of FoxO1,PEPCK,G6Pase,and ApoC-Ⅲin liver tissues of mice in the model group were significantly up-regulated(P<0.01).As compared with the model group,the mRNA and protein levels of FoxO1,PEPCK,G6Pase,and ApoC-Ⅲin the ZJQP group was significantly decreased(P<0.01).Conclusion:ZJQP can improve the glucose and lipid metabolism of T2DM with NAFLD and repair the pathological damage of liver,which may be through regulating the expression of FoxO1,PEPCK,G6Pase,ApoC-Ⅲ-related proteins in liver tissues to achieve the effects of regulating lipid,lowering glucose,and delaying hepatic steatosis.
作者
王一阳
邹俊驹
陆源源
向琴
喻嵘
WANG Yiyang;ZOU Junju;LU Yuanyuan;XIANG Qin;YU Rong(Hunan University of Chinese Medicine,Changsha 410208,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2023年第10期102-109,共8页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金联合基金重点支持项目(U21A20411)
国家自然科学基金项目(82074400,82004185)
湖南省教育厅科技计划项目(20B450,20K094)。
