腺相关病毒介导的腺苷激酶过表达减轻高脂饮食诱导的小鼠葡萄糖耐量异常与肝脏脂肪变性  

Overexpression of adeno-associated virus-mediated adenosine kinase ameliorates high fat diet-induced glucose intolerance and hepatic steatosis in mice

作　　者：吴越[1] 高庆祥 沈洋[1] 屈淑平[2] 程庆保[1] 姜小清[1] WU Yue;GAO Qing-xiang;SHEN Yang;QU Shu-ping;CHENG Qing-bao;JIANG Xiao-qing(Department of Biliary Tract Surgery(Ⅰ),Eastern Hepatobiliary Surgery Hospital,Naval Medical University(Second Military Medical University),Shanghai 200438,China;Department of Hepatic Surgery(Ⅰ),Eastern Hepatobiliary Surgery Hospital,Naval Medical University(Second Military Medical University),Shanghai 200438,China)

机构地区：[1]海军军医大学(第二军医大学)东方肝胆外科医院胆道一科,上海200438 [2]海军军医大学(第二军医大学)东方肝胆外科医院肝外一科,上海200438

出　　处：《第二军医大学学报》2020年第7期769-774,共6页Academic Journal of Second Military Medical University

基　　金：上海市科学技术委员会重点基础项目(13ZR1410200)。

摘　　要：目的探讨特异性上调肝细胞中腺苷激酶(ADK)表达对高脂饮食诱导的小鼠葡萄糖耐量异常与肝脏脂肪变性的影响。方法20只雄性C57BL/6J小鼠随机分为2组,每组10只,其中1组通过尾静脉注射将携带甲状腺素结合球蛋白(TBG)启动子驱动表达ADK的腺相关病毒(AAV8-TBG-ADK)注入小鼠体内(AAV8-TBG-ADK组),另1组注射对照腺相关病毒AAV8-TBG(AAV8-TBG组)。上述2组小鼠接受腺相关病毒注射后,再分别随机分为2个亚组,每亚组5只,分别给予8周普通饮食或高脂饮食。造模过程中检测各组小鼠体质量变化。造模结束后处死小鼠,采用蛋白质印迹法与qRT-PCR检测各组小鼠肝脏ADK表达,葡萄糖耐量试验检测小鼠葡萄糖耐量情况,H-E染色、油红O染色检测肝组织病理改变及脂滴沉积,qRT-PCR检测小鼠肝组织中糖异生相关基因[葡萄糖-6-磷酸酶(G6P)、磷酸烯醇式丙酮酸羧激酶(PEPCK)]和脂肪生成相关基因[胆固醇调节元件结合蛋白1(SREBP-1)、乙酰辅酶A羧化酶1(ACC-1)、脂肪酸合成酶(FAS)、硬脂酰辅酶A脱氢酶1(SCD-1)]的mRNA水平。结果与普通饮食饲喂小鼠相比,高脂饮食可导致小鼠体质量增加、葡萄糖代谢紊乱和肝脏脂质沉积。与注射AAV8-TBG的小鼠相比,注射AAV8-TBG-ADK特异性上调小鼠肝脏ADK的表达后,对普通饮食和高脂饮食饲喂的小鼠体质量均无明显影响,但可减轻高脂饮食导致的葡萄糖耐量异常和肝脏脂质沉积(P均<0.05),并可抑制高脂饮食饲喂小鼠肝脏糖异生与脂肪生成相关基因的表达(P均<0.05)。结论利用腺相关病毒特异性上调肝细胞ADK表达可改善高脂饮食导致的小鼠葡萄糖耐量异常与肝脏脂肪变性。Objective To explore the effects of hepatocyte-specific overexpression of adenosine kinase(ADK)on glucose intolerance and hepatic steatosis induced by high fat diet in mice.Methods Twenty C57BL/6J male mice were randomly divided into two groups(n=10):the mice in one group were injected through tail vein with adeno-associated virus vector carrying thyroxine-binding globulin(TBG)promotor driven expression of ADK(AAV8-TBG-ADK group),while those in the other group were injected with control vector AAV8-TBG(AAV8-TBG group).After adeno-associated virus administration,the mice in the two groups were randomly divided into two subgroups(n=5)to receive normal or high fat diet for the following 8 weeks,and changes in mice body weight were recorded.Mice were sacrificed after modeling.The expressions of ADK in the liver were determined by Western blotting and quantitative real-time polymerase chain reaction(qRT-PCR).The glucose homeostasis was assessed by glucose tolerance test.The pathological changes and lipid deposition of liver tissues were detected by Hematoxylin-Eosin(H-E)staining and oil red O staining,respectively.The mRNA levels of gluconeogenesis related genes(glucose-6-phosphatase[G6P]and phosphoenolpyruvate carboxykinase[PEPCK])and fatty acid synthesis related genes(sterol-regulatory element binding protein-1[SREBP-1],acetyl-CoA carboxylase-1[ACC-1],fatty acid synthase[FAS]and stearoyl-CoA desaturase-1[SCD-1])in liver tissues were analyzed by qRT-PCR.Results Compared with the normal diet subgroup,high fat diet induced weight gain,glucose intolerance and hepatic steatosis.There was no significant difference in the body weight between the AAV8-TBG and AAV8-TBG-ADK groups(P>0.05).Compared with the AAV8-TBG group,AAV-TBG-ADK mediated ADK overexpression significantly ameliorated high fat diet-induced glucose intolerance and hepatic steatosis(all P<0.05),and significantly reduced the mRNA levels of gluconeogenesis related genes and fatty acid synthesis related genes in liver tissues(all P<0.05).Conclusion Hepatocytespec

关 键 词：腺苷激酶 高脂饮食 葡萄糖耐量异常 非酒精性脂肪肝 

分 类 号：R589.2[医药卫生—内分泌]