發(fā)布時(shí)間：2018-03-25 01:04

  本文選題：IGF1　切入點(diǎn)：FoxO1　出處：《安徽醫(yī)科大學(xué)》2015年碩士論文

【摘要】：第一部分高糖下IGF1對(duì)內(nèi)皮細(xì)胞增殖移行影響的研究目的:血管內(nèi)皮功能失調(diào)在糖尿病血管并發(fā)癥中發(fā)揮重要作用。研究發(fā)現(xiàn)糖尿病患者心腦血管病變及周?chē)荛]塞的風(fēng)險(xiǎn)遠(yuǎn)高于非糖尿病患者[1]。臨床研究發(fā)現(xiàn)高血糖與糖尿病血管病變有關(guān),內(nèi)皮細(xì)胞功能失調(diào)可能繼發(fā)于高血糖[2]。胰島素樣生長(zhǎng)因子1(IGF1)是細(xì)胞生長(zhǎng)、分化和凋亡的重要調(diào)節(jié)因子。Fox O(Forkhead box O)是Forkhead轉(zhuǎn)錄因子的一個(gè)亞家族,在哺乳動(dòng)物細(xì)胞周期調(diào)控、凋亡、應(yīng)激以及糖代謝調(diào)節(jié)中起著重要作用。Fox O1在內(nèi)皮細(xì)胞高表達(dá)。近年來(lái)研究證明IGF1具有多種代謝和血管保護(hù)作用,從很多方面直接對(duì)抗內(nèi)皮功能失調(diào)[3],目前關(guān)于IGF1的報(bào)道多關(guān)注于其在神經(jīng)損傷方面的研究,在血管保護(hù)方面的報(bào)道較少,且機(jī)制不清。本研究擬探討IGF1對(duì)高糖環(huán)境中的內(nèi)皮細(xì)胞增殖及移行的保護(hù)作用及其可能機(jī)制。方法:體外培養(yǎng)人臍靜脈內(nèi)皮細(xì)胞(human umbilical vein endothelial cells,HUVEC),倒置顯微鏡下觀察內(nèi)皮細(xì)胞形態(tài)。亞融合狀態(tài)的血管內(nèi)皮細(xì)胞經(jīng)血清饑餓處理24小時(shí)后用于實(shí)驗(yàn)。分為對(duì)照組(5mmol/L Gs)、高糖組(25mmol/L Gs)、高滲組(20mmol/L mannitol+5mmol/L Gs)、高糖+胰島素樣生長(zhǎng)因子-1組(25mmol/L Gs+80ng/m L IGF1)。處理48h后,用四甲基偶氮唑藍(lán)(MTT)比色法檢測(cè)內(nèi)皮細(xì)胞增殖情況,Transwell移行小室觀察細(xì)胞移行情況。實(shí)時(shí)熒光定量PCR測(cè)定Akt m RNA及Fox O1 m RNA水平。免疫組化測(cè)定內(nèi)皮細(xì)胞Akt、p-Akt、Fox O1和p-Fox O1的蛋白表達(dá)情況,并經(jīng)行半定量分析,比較Fox O1/p-Fox O1、Akt/p-Akt的比值。結(jié)果:內(nèi)皮細(xì)胞在高糖組的細(xì)胞存活率、移行率比對(duì)照組明顯降低(P0.01,P0.01),Fox O1 m RNA表達(dá)明顯升高(P0.01),Fox O1/p-Fox O1蛋白表達(dá)增加(P=0.007),Akt/p-Akt比值增加(P0.05)。與高糖組相比,高糖+IGF1組細(xì)胞存活率、移行率明顯增加(P0.05,P0.01),Fox O1 m RNA的表達(dá)明顯增加(P0.05),Fox O1/p-Fox O1蛋白表達(dá)降低(P0.05),Akt/p-Akt蛋白表達(dá)降低(P=0.040);高糖+IGF1+AZD5363組細(xì)胞存活率未見(jiàn)明顯差異(P0.05),移行未增加(P0.05),Fox O1 m RNA的表達(dá)明顯降低(P0.05),Fox O1/p-Fox O1未見(jiàn)明顯差異(P0.05)。五組Akt m RNA的表達(dá)無(wú)差異(F=0.39,P0.05)。結(jié)論:高糖環(huán)境下內(nèi)皮細(xì)胞的增殖移行能力降低與Fox O1的表達(dá)有關(guān)。IGF1能改善高糖對(duì)內(nèi)皮細(xì)胞增殖移行的抑制作用,其機(jī)制可能是IGF1激活A(yù)kt進(jìn)一步調(diào)節(jié)Fox O1,導(dǎo)致Fox O1磷酸化,去磷酸化Fox O1蛋白表達(dá)減少。第二部分中國(guó)2型糖尿病患者血清IGF1與HDL-C的關(guān)系目的:2型糖尿病(T2DM)患者是脂代謝紊亂及動(dòng)脈硬化的高發(fā)人群,關(guān)注IGF1在T2DM人群中的變化及對(duì)血脂的影響,對(duì)于認(rèn)識(shí)胰島素樣生長(zhǎng)因子-1(IGF1)與T2DM患者動(dòng)脈粥樣硬化的關(guān)系非常重要。本文探討中國(guó)T2DM患者患者血清IGF1和血脂水平的關(guān)系。方法:檢測(cè)498例T2DM患者血清IGF1和總膽固醇(TC)、甘油三脂(TG)、低密度脂蛋白膽固醇(LDL-C)、高密度脂蛋白膽固醇(HDL-C)、脂蛋白A(Lp A)和脂蛋白B(Lp B)的水平,并按照第25、75百分位數(shù)將IGF1分為3組即低IGF1組(G1)、中間IGF1組(G2)和高IGF1組(G3)進(jìn)行比較。對(duì)IGF1與HDL-C的相關(guān)性進(jìn)行分析。結(jié)果:與G2組比較,G1組BMI,FINS顯著升高,HDL-C顯著降低(P0.05);G3組HDL-C,2h PBG,FINS,2h PINS,WHR,Hb A1c顯著降低(P0.05),冠心病患病率明顯降低(P0.05)。校正年齡和性別后,血清IGF1水平與HDL-C呈正相關(guān),與年齡,病程,腰圍,FINS,2h PINS,IR負(fù)相關(guān)。以HDL-C為因變量,校正年齡和性別進(jìn)行的逐步多元線性回歸分析中,TG,TC,LDL-C,WHR,2h PBG,IGF1和性別進(jìn)入回歸方程。IGF1每增加2.02 ng/d L,HDL-C增加1.00 mg/d L。結(jié)論:T2DM患者血清IGF1可能是HDL-C的一個(gè)獨(dú)立的正性影響因素,IGF1可能對(duì)2型糖尿病患者心血管疾病的發(fā)生具有保護(hù)性作用。
[Abstract]:Objective to study the first part under high glucose IGF1 transitional effects on the proliferation of endothelial cells: endothelial dysfunction plays an important role in diabetic vascular complications. The study found that the risk of diabetes and cardiovascular disease in patients with peripheral vascular occlusion is much higher than that in non-diabetic patients with [1]. clinical study found that high blood sugar is associated with diabetic vascular disease, endothelial dysfunction may secondary to high blood glucose [2]. insulin-like growth factor 1 (IGF1) is an important regulator of cell growth, differentiation and apoptosis of.Fox O (Forkhead box O) is a subfamily of Forkhead transcription factors in mammalian cell cycle regulation, apoptosis, stress and glucose metabolism plays an important role in the expression of O1 in.Fox endothelial cells. Recent studies show that IGF1 has the metabolism and vascular protection, from many aspects directly against endothelial dysfunction in [3]. Previous reports on IGF1 pay more attention to the study on the nerve injury, in vascular protection reported less, and the mechanism is not clear. This study aims to investigate the protective effect of IGF1 on proliferation of endothelial cells in high glucose environment and migration and its possible mechanism. Methods: human umbilical vein endothelial cells in vitro (human umbilical vein endothelial cells, HUVEC), endothelial cells were observed under inverted microscope. The sub fusion state of vascular endothelial cells by serum starvation for 24 hours after the experiment. Divided into control group (5mmol/L Gs), high glucose group (25mmol/L Gs), hypertonic group (20mmol/L mannitol+5mmol/L Gs), high glucose + insulin like growth factor -1 group (25mmol/L Gs+80ng/m L IGF1). After 48h treatment, with four methyl thiazolyl tetrazolium (MTT) assay of endothelial cell proliferation than color method, Transwell transitional cell were observed shift situation. Real time fluorescent quantitative PCR determination Akt m and RNA Fox O1 m RNA. Immunohistochemical determination of endothelial cells Akt, p-Akt, O1 and p-Fox expression of Fox O1 protein, and analyzed by semi quantitative Fox, O1/p-Fox O1, the ratio of Akt/p-Akt. Results: the survival rate of endothelial cells in high glucose group the cell migration rate than the control group decreased (P0.01, P0.01), the expression of Fox O1 m RNA increased significantly (P0.01), Fox O1/p-Fox increased the expression of O1 (P=0.007), the ratio of Akt/p-Akt increased (P0.05). Compared with the high glucose group, high glucose group +IGF1 cell survival rate, migration rate was increased significantly (P0.05, P0.01), the expression of Fox O1 m RNA significantly increased (P0.05), reduced the expression of Fox O1 protein O1/p-Fox (P0.05), Akt/p-Akt protein expression decreased (P=0.040); high survival rate in +IGF1+AZD5363 group showed no significant difference (P0.05), transitional did not increase the expression of Fox (P0.05), O1 m RNA (P0.05, Fox) significantly decreased O1/p-Fox O1 showed no significant difference (P0 .05). No difference in expression of five groups of Akt m RNA (F=0.39, P0.05). Conclusion:.IGF1 can improve the glucose and reduce the shift inhibitory effect on endothelial cell proliferation the expression of Fox O1 endothelial cell proliferation induced by high glucose migration, the mechanism may be the activation of IGF1 Akt Fox O1 to further regulate. Fox O1 phosphorylation, dephosphorylation of Fox O1 protein expression decreased. The purpose of the second part is the relationship between Chinese in patients with type 2 diabetes, serum IGF1 and HDL-C: type 2 diabetes mellitus (T2DM) patients are at high risk of lipid metabolism and atherosclerosis, changes on IGF1 in the T2DM population and its effect on serum lipid, insulin for understanding insulin-like growth factor -1 (IGF1) and atherosclerosis in T2DM patients is very important. This paper discusses the relationship between China serum IGF1 and lipid levels in patients with T2DM. Methods: 498 T2DM patients serum IGF1 and total cholesterol (TC), glycerol three 鑴，

本文編號(hào)：1660834