血管内皮生长因子、基质细胞衍生因子-1基因对人微血管内皮细胞增殖、迁移、凋亡的影响  

Influence of VEGF,SDF-1 gene on proliferation,migration and apoptosis of human microvascular endothelial cells

作　　者：高静[1] 李晓岚[1] 王敏哲[1] 刘贯英 杨雪松[1] GAO Jing;LI Xiaolan;WANG Minzhe;LIU Guanying;YANG Xuesong(Department of Endocrinology,The Fifth Affiliated Hospital,Xinjiang Medical University,Wulumuqi,Xinjiang Uygur Autonomous Region 830054,China)

机构地区：[1]新疆医科大学第五附属医院内分泌科,新疆维吾尔自治区乌鲁木齐830054

出　　处：《安徽医药》2019年第9期1784-1789,共6页Anhui Medical and Pharmaceutical Journal

基　　金：新疆维吾尔自治区自然科学基金（2014211C129）

摘　　要：目的探讨血管内皮生长因子(VEGF)、基质细胞衍生因子?1(SDF?1)基因对人微血管内皮细胞(HMEC?1)增殖、迁移、凋亡的影响。方法体外培养HMEC?1,以小干扰RNA(siRNA)介导低表达VEGF、SDF?1处理细胞,分组为Ⅰ组(空白对照组)、Ⅱ组(转染非特异性对照组)、Ⅲ组(转染si VEGF组)、Ⅳ组(转染si SDF?1组)。检测各组细胞增殖、迁移、凋亡的情况。West?ern Blot检测细胞中VEGF、SDF?1的表达情况,MTT和流式细胞分析检测HMEC?1增殖和凋亡能力的变化情况,划痕愈合实验检测HMEC?1迁移能力。结果转染了VEGF165?siRNA后,Ⅲ组VEGF165、SDF?1蛋白的表达强度(0.48±0.07)、(0.62±0.08)均明显弱于Ⅰ组(1.92±0.42)、(1.29±0.38)和Ⅱ组(1.87±0.35)、(1.32±0.47),差异有统计学意义(t=9.836,8.279,7.846,8.045,P<0.05);转染了SDF?1?siRNA后,Ⅳ组SDF?1蛋白的表达强度(0.37±0.05)均明显弱于Ⅰ组和Ⅱ组(t=7.381,7.984,P<0.05),差异有统计学意义,而VEGF165蛋白表达变化差异无统计学意义(P>0.05)。Ⅰ组与Ⅱ组比较,VEGF165、SDF?1蛋白表达水平差异无统计学意义(P>0.05)。经siRNA转染后,Ⅲ组、Ⅳ组细胞均出现增殖抑制率(42.37±1.25)%、(29.15±1.32)%和凋亡率(21.6±1.8)%、(16.8±1.6)%明显增高,与Ⅰ组、Ⅱ组增殖抑制率0.00%、(0.24±0.02)%和凋亡率(4.9±0.4)%、(5.2±0.5)%比较,差异有统计学意义(P<0.05)。但Ⅲ组和Ⅳ组之间比较,Ⅲ组细胞增殖抑制率和凋亡率增高更显著,差异有统计学意义(t=6.217,5.487,P<0.05)。Ⅰ组与Ⅱ组比较,细胞增殖抑制率和凋亡率均差异无统计学意义(P>0.05)。细胞迁移实验中,Ⅰ组、Ⅱ组、Ⅲ组、Ⅳ组迁移距离分别为(579.65±11.59)μm、(553.76±9.47)μm、(234.72±10.28)μm、(318.36±9.95)μm,Ⅲ组和Ⅳ组细胞明显少于Ⅰ组和Ⅱ组,差异有统计学意义(t=10.972,9.894,8.426,7.904,P<0.05),而Ⅰ组与Ⅱ组之间,差异无统计学意义(P>0.05),其中Ⅲ组和Ⅳ组之间比较,Ⅲ组细胞明显少于Ⅳ组,差异有�Objective To study the influence of vascular endothelial growth factor(VEGF),stromal cell?derived factor?1(SDF?1)gene on proliferation,migration and apoptosis of human microvascular endothelial cells(HMEC?1).Methods HMEC?1 was cul?tured in vitro,and VEGF and SDF?1 cells were mediated by small interfering RNA(siRNA)and assigned into control group(I group),scramble group(Ⅱgroup),transfection siVEGF group(Ⅲgroup)and transfection siSDF?1 group(IV group).The prolifera?tion,migration,and apoptosis of each group of cells were examined.The alteration of VEGF,SDF?1 protein expression were detected by western blot.The proliferation ability was determined by MTT and the cell apoptosis were determined by flow cytometry analysis.The cell migrating ability was detected by transwell migration assay.Results After the transfection of VEGF165?siRNA,VEGF165 and SDF?1 protein expression ofⅢgroup(0.48±0.07),(0.62±0.08)were significantly lower than(1.92±0.42),(1.29±0.38)of I group and(1.87±0.35),(1.32±0.47)of II group,and the difference was statistically significant(t=9.836,8.279,9.836,8.279,P<0.05).After transfection SDF?1?siRNA,SDF?1 protein expression of IV group(0.37±0.05)was significantly lower than that of I andⅡgroup,and the difference was statistically significant(t=7.381,7.984,P<0.05),but the difference of VEGF165 protein expres?sion was not statistically significant(P>0.05).The differences of VEGF165 and SDF?1expression levels were no statistically significant between I group and II group(P>0.05).After siRNA transfection,the cell proliferation inhibition,and apoptosis rate of Ⅲgroup[(42.37±1.25)%,(21.6±1.8)%]and IV group[(29.15±1.32)%,(16.8±1.6%)]were significantly higher than[0.00%,(4.9±0.4)%]I group and[(0.24±0.02)%,(5.2±0.5)%]Ⅱ group with statistical significance(P<0.05).Compared with IV groups,thecell proliferation inhibition and apoptosis rate of Ⅲ group was increased more significantly(t=6.217,5.487,P<0.05).There wereno obvious difference of cell proliferation inhibition and apoptosis rate b

关 键 词：血管内皮生长因子 基质细胞衍生因子-1 糖尿病视网膜病变 人微血管内皮细胞 RNA干扰技术 

分 类 号：R73[医药卫生—肿瘤]