| 高静,李晓岚,王敏哲,等.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[J].安徽医药,2019,23(9):1784-1789. |
| 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 |
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| DOI：10.3969/j.issn.1009-6469.2019.09.021 |
| 中文关键词: 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 |
| 英文关键词: Vascular endothelial growth factor Stromal cell-derived factor-1 Diabetic retinopathy Human microvascular endothelial cells RNA interference |
| 基金项目:新疆维吾尔自治区自然科学基金(2014211C129) |
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| 摘要点击次数: 1654 |
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| 中文摘要: |
| 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。Western 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，差异有统计学意义(t＝5.907，P＜0.05)。结论 VEGF、SDF-1基因均参能促进HMEC-1的增殖和迁移能力，并抑制HMEC-1的凋亡水平，从而可能在糖尿病血管病变发生发展中发挥作用，且在此过程中VEGF对SDF-1表达具有调节机制。 |
| 英文摘要: |
| 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 cultured in vitro，and VEGF and SDF-1 cells were mediated by small interfering RNA(siRNA) and assigned into control group(I group)，scramble group(Ⅱ group)，transfection siVEGF group(Ⅲ group) and transfection siSDF-1 group(IV group).The proliferation，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 of Ⅲ group(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 and Ⅱ group，and the difference was statistically significant(t＝7.381，7.984，P＜0.05)，but the difference of VEGF165 protein expression 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 Ⅲ group ［(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，the cell proliferation inhibition and apoptosis rate of Ⅲ group was increased more significantly(t＝6.217，5.487，P＜0.05).There were no obvious difference of cell proliferation inhibition and apoptosis rate between I and Ⅱ group(P＞0.05).In cell migration experiment，the migration distance of I，Ⅱ，Ⅲ，IV group were(579.65±11.59) μm，(553.76±9.47) μm，(234.72±10.28) μm and(318.36±9.95) μm，respectively.The number of cells in Ⅲ group and IV group was significantly lower than that of I and Ⅱ group(t＝10.972，9.894，10.972，9.894，P＜0.05).There was no statistically significant difference between I and Ⅱ group(P＞0.05)，and the migration distance of Ⅲ group was significantly less than that of IV group(t＝5.907，P＜0.05).Conclusion VEGF and SDF-1 could promote the proliferation and migration of HMEC-1，and inhibit the apoptosis of HMEC-1，which may play a role in the development of diabetes vascular lesions occur，and VEGF may modulate the expressions of SDF-1 in this process. |
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