高糖下IGF1对内皮细胞增殖及移行的影响

发布时间：2018-03-25 01:04

  本文选题：IGF1　切入点：FoxO1　出处：《安徽医科大学》2015年硕士论文

【摘要】：第一部分高糖下IGF1对内皮细胞增殖移行影响的研究目的:血管内皮功能失调在糖尿病血管并发症中发挥重要作用。研究发现糖尿病患者心脑血管病变及周围血管闭塞的风险远高于非糖尿病患者[1]。临床研究发现高血糖与糖尿病血管病变有关,内皮细胞功能失调可能继发于高血糖[2]。胰岛素样生长因子1(IGF1)是细胞生长、分化和凋亡的重要调节因子。Fox O(Forkhead box O)是Forkhead转录因子的一个亚家族,在哺乳动物细胞周期调控、凋亡、应激以及糖代谢调节中起着重要作用。Fox O1在内皮细胞高表达。近年来研究证明IGF1具有多种代谢和血管保护作用,从很多方面直接对抗内皮功能失调[3],目前关于IGF1的报道多关注于其在神经损伤方面的研究,在血管保护方面的报道较少,且机制不清。本研究拟探讨IGF1对高糖环境中的内皮细胞增殖及移行的保护作用及其可能机制。方法:体外培养人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC),倒置显微镜下观察内皮细胞形态。亚融合状态的血管内皮细胞经血清饥饿处理24小时后用于实验。分为对照组(5mmol/L Gs)、高糖组(25mmol/L Gs)、高渗组(20mmol/L mannitol+5mmol/L Gs)、高糖+胰岛素样生长因子-1组(25mmol/L Gs+80ng/m L IGF1)。处理48h后,用四甲基偶氮唑蓝(MTT)比色法检测内皮细胞增殖情况,Transwell移行小室观察细胞移行情况。实时荧光定量PCR测定Akt m RNA及Fox O1 m RNA水平。免疫组化测定内皮细胞Akt、p-Akt、Fox O1和p-Fox O1的蛋白表达情况,并经行半定量分析,比较Fox O1/p-Fox O1、Akt/p-Akt的比值。结果:内皮细胞在高糖组的细胞存活率、移行率比对照组明显降低(P0.01,P0.01),Fox O1 m RNA表达明显升高(P0.01),Fox O1/p-Fox O1蛋白表达增加(P=0.007),Akt/p-Akt比值增加(P0.05)。与高糖组相比,高糖+IGF1组细胞存活率、移行率明显增加(P0.05,P0.01),Fox O1 m RNA的表达明显增加(P0.05),Fox O1/p-Fox O1蛋白表达降低(P0.05),Akt/p-Akt蛋白表达降低(P=0.040);高糖+IGF1+AZD5363组细胞存活率未见明显差异(P0.05),移行未增加(P0.05),Fox O1 m RNA的表达明显降低(P0.05),Fox O1/p-Fox O1未见明显差异(P0.05)。五组Akt m RNA的表达无差异(F=0.39,P0.05)。结论:高糖环境下内皮细胞的增殖移行能力降低与Fox O1的表达有关。IGF1能改善高糖对内皮细胞增殖移行的抑制作用,其机制可能是IGF1激活Akt进一步调节Fox O1,导致Fox O1磷酸化,去磷酸化Fox O1蛋白表达减少。第二部分中国2型糖尿病患者血清IGF1与HDL-C的关系目的:2型糖尿病(T2DM)患者是脂代谢紊乱及动脉硬化的高发人群,关注IGF1在T2DM人群中的变化及对血脂的影响,对于认识胰岛素样生长因子-1(IGF1)与T2DM患者动脉粥样硬化的关系非常重要。本文探讨中国T2DM患者患者血清IGF1和血脂水平的关系。方法:检测498例T2DM患者血清IGF1和总胆固醇(TC)、甘油三脂(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)、脂蛋白A(Lp A)和脂蛋白B(Lp B)的水平,并按照第25、75百分位数将IGF1分为3组即低IGF1组(G1)、中间IGF1组(G2)和高IGF1组(G3)进行比较。对IGF1与HDL-C的相关性进行分析。结果:与G2组比较,G1组BMI,FINS显著升高,HDL-C显著降低(P0.05);G3组HDL-C,2h PBG,FINS,2h PINS,WHR,Hb A1c显著降低(P0.05),冠心病患病率明显降低(P0.05)。校正年龄和性别后,血清IGF1水平与HDL-C呈正相关,与年龄,病程,腰围,FINS,2h PINS,IR负相关。以HDL-C为因变量,校正年龄和性别进行的逐步多元线性回归分析中,TG,TC,LDL-C,WHR,2h PBG,IGF1和性别进入回归方程。IGF1每增加2.02 ng/d L,HDL-C增加1.00 mg/d L。结论:T2DM患者血清IGF1可能是HDL-C的一个独立的正性影响因素,IGF1可能对2型糖尿病患者心血管疾病的发生具有保护性作用。
[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，

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