重组腺相关病毒介导的VLDLR基因对2型糖尿病大鼠脂代谢紊乱的作用  

Effect of rAAV-mediated VLDLR gene on the hyperlipidemia in the model of type 2 diabetes mellitus

作　　者：刘永健[1,2] 张木勋[1] 张建华[1] 徐咏平[1] 孙婷婷[1] 杨雁[1] 李昌林[1] 袁刚[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院内分泌科,湖北武汉430030 [2]武汉市普爱医院内分泌科

出　　处：《中国病理生理杂志》2009年第2期344-350,共7页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30400220)

摘　　要：目的:探索增加极低密度脂蛋白受体(VLDLR)基因表达对2型糖尿病大鼠脂代谢紊乱和动脉粥样斑块的影响。方法:构建携带人VLDLR基因的重组腺相关病毒载体(rAAV-VLDLR)。高脂高糖饮食8周后尾静脉注射小剂量链脲佐菌素(STZ)造2型糖尿病动物模型,VLDLR治疗组大鼠注射rAAV-VLDLR,糖尿病对照组注射rAAV-0。RT-PCR及Western blotting分别检测大鼠骨骼肌、心脏、主动脉、肝脏、脂肪VLDLR mRNA及蛋白表达水平。检测VLDLR基因对血糖、胰岛素、稳态模型胰岛素抵抗指数(HOMA-IR)、甘油三酯(TG)、胆固醇(TC)、脂蛋白脂酶(LPL)活性及主动脉粥样斑块的影响。结果:rAAV-0组大鼠骨骼肌、主动脉、脂肪VLDLR mRNA及蛋白水平低于正常组,VLDLR治疗组VLDLR mRNA及蛋白水平较rAAV-0组有不同程度增加。VLDLR治疗后4、8周的TG及4、8、12周的TC明显降低(P<0.05),8周HOMA-IR明显降低(P<0.05),LPL活性明显增强(P<0.05),主动脉内膜损伤减轻。结论:增加VLDLR基因表达对改善2型糖尿病大鼠脂代谢紊乱效果显著,并可在此基础上减轻主动脉粥样斑块程度。AIM ： To investigate the effect of very low density lipoprotein receptor （VLDLR） gene therapy on the hyperlipidemia and atherosclerosis in type 2 diabetes rats. METHODS ： Rats in type 2 diabetic group was fed with high fat and high glucose diet, after 8 weeks STZ （streptozotocin, 25 mg/kg body weight） was injected via the tail vein, while the normal control group was fed with normal diet. Recombinant adeno - associated virus （rAAV） mediated VLDLR gene or rAAV -0 was introduced into the diabetic rats. The expressions of VLDLR mRNA and protein were detected by reverse transcription polymerase chain reaction （RT - PCR） and Western blotting in muscle, heart, aorta, liver and adipose tissues. The fasting blood glucose, fasting insulin, homeostasis model assessment of insulin resistance （ HOMA - IR）, triglyceride （TG）, total cholesterol （TC） and lipoprotein lipase activity were measured after VLDLR treatment. The atherosclerotic plaque was determined bY histochemical method. RESULTS： The VLDLR mRNA and protein levels in muscle, aorta, adipose were lower in rAAV - 0 rats than those in normal control rats. Compared with the rAAV - 0 group, the VLDLR mRNA and protein expressions increased in rAAV- VLDLR group. The triglyceride （4 weeks, 8 weeks） , cholesterol （4 weeks, 8 weeks, 12 weeks） and HOMA - IR （8 weeks） in rAAV - VLDLR group were significantly lower than those in rAAV - 0 group after treatment （P 〈 0. 05 ）. In rAAV - VLDLR group, the plasma LPL activity was significantly enhanced than that in rAAV - 0 group （P 〈 0. 05 ）. The endothelial damage of the aortas in rAAV - VLDLR rats was less severe than that in rAAV - 0 rats. CONCLUSION ： The recombinant adeno - associated virus mediated VLDLR gene therapy improves the hyperlipidemia and attenuates the development of atherosclerosis in type 2 diabetic rats.

关 键 词：受体 LDL 基因疗法 腺病毒 糖尿病 高脂血症 动脉硬化 

分 类 号：R363[医药卫生—病理学]