Angptl3基因在SD大鼠体内组织表达差异与血脂、Ⅱ型糖尿

发布时间：2018-01-13 06:03

本文关键词：Angptl3基因在SD大鼠体内组织表达差异与血脂、Ⅱ型糖尿病、IgA肾病关系的探讨　出处：《重庆医科大学》2007年硕士论文　论文类型：学位论文

【摘要】： 目的:通过制作高脂血症、Ⅱ型糖尿病并发高脂血症、IgA肾病并发高脂血症大鼠模型,采用Realtime PCR(SYBR GreenⅠ)检测血管生成素样蛋白3(angiopoietin-related protein 3,ANGPTL3)和脂蛋白脂肪酶(Lipoprotein lipase,LPL)在脑、肺、心、肝、胃、肾、脂肪等7种组织中mRNA的表达水平,探讨Angptl 3组织表达特异性,分析Angptl 3、LPL在大鼠体内上述各组织表达水平与血脂水平、Ⅱ型糖尿病、IgA肾病的相关性,为进一步研究Angptl 3参与调节脂代谢的机制奠定基础。方法:以SD大鼠肝组织总RNA为模板,通过RT-PCR及T-A克隆,将GAPDH、Angptl3、LPL等基因片段装载到质粒上,并以此为模板绘制标准曲线。将40只健康雄性大鼠随机分为正常对照组(Normal Control,NC)、高脂血症组(Hyperlipidemia,HL)、Ⅱ型糖尿病组(Diabetes Mellitus,DM)、IgA肾病组(IgA Nephropathy,IN)等4组,10只/组。正常对照组给予普通饮食,其余三组给予高脂饮食以诱导高脂血症。DM组大鼠按30mg/kg单剂量腹腔注射STZ (Streptozotocin) ; IN组联合应用牛血清白蛋白(Bovine serum albumin,BSA)、蓖麻油、CCl4、脂多糖(Lipopolysaccharide,LPS)。饲养3个月后处死动物, Realtime PCR检测上述7种组织内GAPDH、Angptl3、LPL的mRNA表达水平。结果:三条标准曲线相关系数分别为:0.9976、0.9991、0.9996,反应效率分别为:0.9793、0.8623、0.8967,溶解曲线呈单一峰型。GAPDH在各组织均有稳定表达;Angptl3仅在肝、肾组织中表达,表达水平DM组HL组IN组NC组。LPL主要在心、脂肪等组织表达,表达水平HL组IN组DM组NC组。结论:在血脂水平不同、Ⅱ型糖尿病、IgA肾病等不同影响因素下,Angptl3仅在肝、肾组织表达,组织表达特异性并未改变。Angptl3的表达水平与血清胆固醇水平、胰岛素缺乏等存在密切联系,IgA肾病并未影响到Angptl3的表达水平。LPL表达水平与血脂水平、糖尿病状态等密切相关,Angptl3是否上调LPL的表达,尚需进一步研究。
[Abstract]:Objective: through the production of hyperlipidemia, diabetic hyperlipidemia, IgA nephropathy complicated with hyperlipidemia rat model, using Realtime PCR (SYBR Green 1) detection of angiopoietin like protein 3 (angiopoietin-related protein 3, ANGPTL3) and lipoprotein lipase (Lipoprotein lipase, LPL) in the brain, lung, heart liver, stomach, kidney, fat, the expression level of mRNA in 7 different tissues, the expression of Angptl and tissue specificity of 3, the analysis of Angptl 3, LPL level and blood lipid level in rats. The expression of tissue type II diabetic nephropathy associated IgA, for the further study of Angptl 3 mechanisms involved in the regulation of lipid metabolism to lay the foundation.
Methods: total RNA of liver tissues of rats with SD as template by RT-PCR and T-A GAPDH, Angptl3, cloning, loading LPL gene fragment into plasmid, and as a standard curve. A total of 40 healthy male rats were randomly divided into normal control group (Normal, Control, NC), hyperlipidemia group (Hyperlipidemia, HL), diabetes group (Diabetes, Mellitus, DM), IgA (IgA Nephropathy, IN nephropathy group) 4 groups, 10 rats in each group. The control group was given normal diet, the other three groups were given high fat diet to induce hyperlipidemia.DM rats by a single dose of kg 30mg/ intraperitoneal injection of STZ (Streptozotocin); IN group combined with bovine albumin serum (Bovine serum, albumin, BSA), castor oil, CCl4, lipopolysaccharide (Lipopolysaccharide, LPS). The animals were sacrificed 3 months after the animal, Realtime PCR detection of the 7 tissues of GAPDH, Angptl3, the expression level of LPL mRNA.
Results: the correlation coefficient of three standard curves were: 0.9976,0.9991,0.9996, the reaction efficiency is 0.9793,0.8623,0.8967, dissolution curve expression was single peak type.GAPDH in each tissue was stable; Angptl3 only in the liver, renal tissue, the expression level of DM group HL group IN group NC group.LPL in the heart and adipose tissue expression. The expression level of HL group IN group DM group NC group.
Conclusion: the different lipid levels in type II diabetic nephropathy, IgA and other different factors, Angptl3 only in the liver, kidney tissue expression, tissue specific expression did not change the expression of.Angptl3 and serum cholesterol levels, insulin deficiency are closely linked, IgA nephropathy did not affect the expression level of.LPL Angptl3 expression level and lipids level the closely related diabetes status, whether Angptl3 increase the expression of LPL still need further study.

【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R-332;R587.1;R692.3

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