| 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。 |
| 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 |
| DOI：10.16781/j.0258-879x.2020.07.0769 |
| 投稿时间：2019-12-09修订日期：2020-04-15 |
| 基金项目:上海市科学技术委员会重点基础项目（13ZR1410200）. |
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| Overexpression of adeno-associated virus-mediated adenosine kinase ameliorates high fat diet-induced glucose intolerance and hepatic steatosis in mice |
| WU Yue1,GAO Qing-xiang1,SHEN Yang1,QU Shu-ping2,CHENG Qing-bao1,JIANG Xiao-qing1* |
(1. Department of Biliary Tract Surgery(Ⅰ), Eastern Hepatobiliary Surgery Hospital, Naval Medical University(Second Military Medical University), Shanghai 200438, China;
2. Department of Hepatic Surgery(Ⅰ), Eastern Hepatobiliary Surgery Hospital, Naval Medical University(Second Military Medical University), Shanghai 200438, China
*Corresponding author) |
| Abstract: |
| 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 Hepatocyte-specific overexpression of ADK can improve high fat diet-induced glucose intolerance and hepatic steatosis in mice. |
| Key words: adenosine kinase high fat diet glucose intolerance non-alcoholic fatty liver |
