脂联素基因表达载体的构建及体外表达研究

发布时间：2018-08-24 09:44

【摘要】：随着脂肪细胞因子的不断发现,目前认为,脂肪组织已不再是仅供能量储备的器官,它是一个具有多种内分泌功能的内分泌器官。脂联素(Adiponectin,APN)是由分化的脂肪细胞特异性分泌的一种蛋白质,在体内和体外实验中被证实具有增加脂肪酸氧化、改善胰岛素抵抗、抗动脉粥样硬化等作用。在临床上,低 APN 血症与肥胖、2 型糖尿病及心血管疾病密切相关。Kubota 等人研究发现,杂合子小鼠(adipo+/-)与野生型小鼠相比表现出微弱的胰岛素抵抗,而纯合子小鼠(adipo-/-)表现出中度的胰岛素抵抗和轻微的糖耐量异常;且 APN敲除纯合子小鼠(adipo-/-)在给予外源性(机械性)血管损伤后,其动脉内膜增厚的发生率要比野生型小鼠高两倍以上,而感染能产生APN 的腺病毒之后却能完全扭转动脉内膜的过度增生。因此,APN 在体内可能有改善胰岛素抵抗和抗动脉粥样硬化的作用。APN 基因治疗可能为肥胖、2 型糖尿病和动脉粥样硬化等疾病开辟新的治疗途径。2型糖尿病常常与胰岛素抵抗、高血压、血脂紊乱及慢性低度炎症反应(通称代谢综合征,MS)同时存在,上述因素是动脉粥样硬化危险因素,他们之间可能存在共同的发病机制。本研究旨在构建 APN 基因表达载体,研究其在体外培养的表达情况,为进一步研究 APN 基因转移抗糖尿病、抗动脉粥样硬化作用打下基础。目的:构建人 APN(humanAPN, hAPN)基因的质粒表达载体,并检测其体外转染后能否在真核细胞中表达,探讨该 APN 基因表达载体的构建是否成功。
[Abstract]:With the continuous discovery of adipocyte factors, it is believed that adipose tissue is no longer an organ that only supplies energy reserve, but an endocrine organ with many endocrine functions. Adiponectin (Adiponectin,APN) is a protein secreted by differentiated adipocytes. It has been proved in vivo and in vitro to increase the oxidation of fatty acids, improve insulin resistance and anti-atherosclerosis. Clinically, hypoglycemia is closely associated with obesity, type 2 diabetes and cardiovascular disease. Studies in heterozygous mice (adipo / -) show weak insulin resistance compared with wild-type mice. Homozygous mice (adipo-/-) showed moderate insulin resistance and mild glucose tolerance abnormality, and APN knockout homozygous mice (adipo-/-) were exposed to exogenous (mechanical) vascular injury. The incidence of intimal thickening was more than twice as high as that of wild-type mice, but the adenovirus that produced APN could completely reverse the intimal hypertrophy. Therefore, APN may improve insulin resistance and anti-atherosclerosis in vivo. APN gene therapy may open up a new therapeutic pathway for obesity, type 2 diabetes mellitus and atherosclerosis, type 2 diabetes mellitus and insulin resistance. Hypertension, dyslipidemia and chronic low-grade inflammation (commonly known as metabolic syndrome) occur simultaneously. These factors are risk factors for atherosclerosis and may have common pathogenesis among them. The purpose of this study was to construct a APN gene expression vector and to study its expression in vitro, and to lay a foundation for further study on the anti-diabetes and anti-atherosclerosis effects of APN gene transfer. Aim: to construct a plasmid expressing human APN (humanAPN, hAPN) gene and detect whether it can be expressed in eukaryotic cells after transfection in vitro.
【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R346

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