左归降糖清肝方通过调控FoxO1/MTP/APOB信号通路改善2型糖尿病合并非酒精性脂肪肝糖脂代谢  

Zuogui Jiangtang Qinggan Formula improves glucolipid metabolism in type 2 diabetes mellitus complicated with non-alcoholic fatty liver disease by regulating FoxO1/MTP/APOB signaling pathway

作　　者：项逸欣 黄雅兰 周敏[1] 邹俊驹 刘秀[1] 刘子毓[1] 肖凡[1] 喻嵘[1] 向琴[1] XIANG Yi-xin;HUANG Ya-lan;ZHOU Min;ZOU Jun-ju;LIU Xiu;LIU Zi-yu;XIAO Fan;YU Rong;XIANG Qin(Hunan University of Chinese Medicine,Changsha 410208,China)

机构地区：[1]湖南中医药大学,湖南长沙410208

出　　处：《中国中药杂志》2023年第16期4438-4445,共8页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(82004185,82074400,U21A20411);湖南省教育厅科技计划项目(20B450,20K094);湖南省大学生创新创业训练计划项目(2021-2452);湖南中医药大学2020年大学生创新创业训练计划项目。

摘　　要：旨在探讨左归降糖清肝方对2型糖尿病(type 2 diabetes mellitus,T2DM)合并非酒精性脂肪肝(non-alcoholic fatty liver disease,NAFLD)糖脂代谢的影响及作用机制。以MKR小鼠(T2DM小鼠)为研究对象,采用高脂饮食诱导NAFLD,建立T2DM合并NAFLD模型,随机将40只小鼠分为模型组,二甲双胍组(0.067 g·kg^(-1)),左归降糖清肝方高、低剂量组(29.64、14.82 g·kg^(-1)),每组10只,以同龄FVB小鼠作为正常组。除正常组和模型组外,其他各组连续灌胃给药4周后采集血清和肝组织标本,检测空腹血糖(fasting blood glucose,FBG)、空腹胰岛素(fasting lisulin,FINS)水平;采用GPO-PAP单试剂微板法检测总胆固醇(total cholesterol,TC)、甘油三酯(triglyceride,TG)、低密度脂蛋白(low-density lipoprotein,LDL)水平;酶联免疫吸附测定法(enzyme linked immunosorbent assay,ELISA)检测极低密度脂蛋白(very low-density lipoprotein,VLDL)水平;赖式微板法检测丙氨酸氨基转移酶(alanine aminotransferase,ALT)和天冬氨酸氨基转移酶(aspartate aminotransferase,AST)水平;通过苏木素-伊红(hematoxylin-eosin,HE)染色和油红O染色观察肝脏病理学形态变化;采用实时荧光定量聚合酶链式反应(real-time PCR)和蛋白免疫印迹法(Western blot)检测肝脏中叉头框转录因子O亚族1(forkhead box transcription factor O1,FoxO1)、微粒体甘油三酯转移蛋白(microsomal triglyceride transfer protein,MTP)和载脂蛋白B(apolipoproteins B,APOB)的基因和蛋白表达情况。结果显示,高剂量左归降糖清肝方能显著降低T2DM合并NAFLD小鼠的FBG、FINS水平(P<0.05,P<0.01),改善葡萄糖耐受和胰岛素抵抗(P<0.05,P<0.01);并能缓解高脂饮食导致的肝脏损伤,表现在改善T2DM合并NAFLD小鼠肝脏脂肪变性程度,显著降低血清中TC、TG、LDL、VLDL、ALT及AST水平(P<0.05,P<0.01);同时研究发现,高剂量左归降糖清肝方干预后肝脏中FoxO1、MTP和APOB的基因和蛋白表达显著下调(P<0.05,P<0.01)。上述研究表明,This study aimed to investigate the effect and mechanism of Zuogui Jiangtang Qinggan Formula(ZGJTQG)on the glucolipid metabolism of type 2 diabetes mellitus(T2DM)complicated with non-alcoholic fatty liver disease(NAFLD).NAFLD was induced by a high-fat diet(HFD)in MKR mice(T2DM mice),and a model of T2DM combined with NAFLD was established.Forty mice were randomly divided into a model group,a metformin group(0.067 g·kg^(-1)),and high-and low-dose ZGJTQG groups(29.64 and 14.82 g·kg^(-1)),with 10 mice in each group.Ten FVB mice of the same age were assigned to the normal group.Serum and liver tissue specimens were collected from mice except for those in the normal and model groups after four weeks of drug administration by gavage,and fasting blood glucose(FBG)and fasting insulin(FINS)levels were measured.The levels of total cholesterol(TC),triglyceride(TG),and low-density lipoprotein(LDL)were detected by the single reagent GPO-PAP method.Very low-density lipoprotein(VLDL)was detected by enzyme-linked immunosorbent assay(ELISA).Alanine aminotransferase(ALT)and aspartate aminotransferase(AST)were determined by the Reitman-Frankel assay.The pathological changes in the liver were observed by hematoxylin-eosin(HE)staining and oil red O staining.Real-time fluorescence-based quantitative polymerase chain reaction(real-time PCR)and Western blot were adopted to detect the mRNA and protein expression of forkhead transcription factor O1(FoxO1),microsomal triglyceride transfer protein(MTP),and apolipoprotein B(APOB)in the liver.The results showed that high-dose ZGJTQG could significantly reduce the FBG and FINS levels(P<0.05,P<0.01),improve glucose tolerance and insulin resistance(P<0.05,P<0.01),alleviate the liver damage caused by HFD which was reflected in improving liver steatosis,and reduce the serum levels of TC,TG,LDL,VLDL,ALT,and AST(P<0.05,P<0.01)in T2DM mice combined with NAFLD.The findings also revealed that the mRNA and protein expression of FoxO1,MTP,and APOB in the liver was significantly down-regulated after the

关 键 词：左归降糖清肝方 2型糖尿病 非酒精性脂肪肝 FOXO1 糖脂代谢 

分 类 号：R285.5[医药卫生—中药学]