本文關(guān)鍵詞：Angptl3基因在SD大鼠體內(nèi)組織表達(dá)差異與血脂、Ⅱ型糖尿病、IgA腎病關(guān)系的探討　出處：《重慶醫(yī)科大學(xué)》2007年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： Angptl3 高脂血癥 Ⅱ型糖尿病 IgA腎病

【摘要】： 目的:通過制作高脂血癥、Ⅱ型糖尿病并發(fā)高脂血癥、IgA腎病并發(fā)高脂血癥大鼠模型,采用Realtime PCR(SYBR GreenⅠ)檢測(cè)血管生成素樣蛋白3(angiopoietin-related protein 3,ANGPTL3)和脂蛋白脂肪酶(Lipoprotein lipase,LPL)在腦、肺、心、肝、胃、腎、脂肪等7種組織中mRNA的表達(dá)水平,探討Angptl 3組織表達(dá)特異性,分析Angptl 3、LPL在大鼠體內(nèi)上述各組織表達(dá)水平與血脂水平、Ⅱ型糖尿病、IgA腎病的相關(guān)性,為進(jìn)一步研究Angptl 3參與調(diào)節(jié)脂代謝的機(jī)制奠定基礎(chǔ)。 方法:以SD大鼠肝組織總RNA為模板,通過RT-PCR及T-A克隆,將GAPDH、Angptl3、LPL等基因片段裝載到質(zhì)粒上,并以此為模板繪制標(biāo)準(zhǔn)曲線。將40只健康雄性大鼠隨機(jī)分為正常對(duì)照組(Normal Control,NC)、高脂血癥組(Hyperlipidemia,HL)、Ⅱ型糖尿病組(Diabetes Mellitus,DM)、IgA腎病組(IgA Nephropathy,IN)等4組,10只/組。正常對(duì)照組給予普通飲食,其余三組給予高脂飲食以誘導(dǎo)高脂血癥。DM組大鼠按30mg/kg單劑量腹腔注射STZ (Streptozotocin) ; IN組聯(lián)合應(yīng)用牛血清白蛋白(Bovine serum albumin,BSA)、蓖麻油、CCl4、脂多糖(Lipopolysaccharide,LPS)。飼養(yǎng)3個(gè)月后處死動(dòng)物, Realtime PCR檢測(cè)上述7種組織內(nèi)GAPDH、Angptl3、LPL的mRNA表達(dá)水平。 結(jié)果:三條標(biāo)準(zhǔn)曲線相關(guān)系數(shù)分別為:0.9976、0.9991、0.9996,反應(yīng)效率分別為:0.9793、0.8623、0.8967,溶解曲線呈單一峰型。GAPDH在各組織均有穩(wěn)定表達(dá);Angptl3僅在肝、腎組織中表達(dá),表達(dá)水平DM組HL組IN組NC組。LPL主要在心、脂肪等組織表達(dá),表達(dá)水平HL組IN組DM組NC組。 結(jié)論:在血脂水平不同、Ⅱ型糖尿病、IgA腎病等不同影響因素下,Angptl3僅在肝、腎組織表達(dá),組織表達(dá)特異性并未改變。Angptl3的表達(dá)水平與血清膽固醇水平、胰島素缺乏等存在密切聯(lián)系,IgA腎病并未影響到Angptl3的表達(dá)水平。LPL表達(dá)水平與血脂水平、糖尿病狀態(tài)等密切相關(guān),Angptl3是否上調(diào)LPL的表達(dá),尚需進(jìn)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.

【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類號(hào)】：R-332;R587.1;R692.3

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本文編號(hào)：1417752