低氧对人微血管内皮细胞HIF-1α表达及生物学行为的影响

发布时间：2018-10-12 21:11

【摘要】：下肢静脉曲张目前是一种常见疾病,在我国发病率较高,严重影响人民生活质量,因此对静脉性溃疡形成的机制研究有重大的意义。研究目的:研究低氧对人皮肤微血管内皮细胞(human dermal microvascular endothelial cells,HDMEC)迁移、凋亡及HIF-1表达的影响。研究方法:培养人微血管内皮细胞,分为正常氧组(对照组,采取常规培养)和低氧组(实验组,CoCl_2模拟化学缺氧)。CCK-8细胞生存实验确定合适处理浓度(200μmol/L CoCl_2),划痕实验检测细胞迁移能力,流式细胞技术观察细胞凋亡及周期变化;应用RT-PCR和Western Blotting检测HIF-1α、HIF-1β、VEGF、iNOS基因m RNA和蛋白表达。研究结果:与对照组比较,低氧组细胞迁移能力增加(P0.05),凋亡增多(P0.05),均呈时间依赖性。低氧组HIF-1α、HIF-1β、VEGF、iNOS基因mRNA和蛋白表达均增加(P值均小于0.05),具有一定时间依赖性。研究结论:低氧能增强人皮肤微血管内皮细胞迁移能力,并促进其凋亡,其可能机制与HIF-1α及其下游基因表达升高有关。
[Abstract]:Varicose veins of the lower extremity is a common disease at present. The incidence of varicose veins in China is high, which seriously affects the quality of life of the people, so it is of great significance to study the mechanism of the formation of venous ulcer. Aim: to study the effects of hypoxia on the migration, apoptosis and HIF-1 expression of human microvascular endothelial cells (human dermal microvascular endothelial cells,HDMEC). Methods: human microvascular endothelial cells were cultured and divided into normal oxygen group (control group). The normal culture and hypoxia group (experimental group, CoCl_2 simulated chemical hypoxia) were used. The suitable concentration of CCK-8 cell survival was determined (200 渭 mol/L CoCl_2), the cell migration ability was detected by scratch test, and cell apoptosis and cell cycle change were observed by flow cytometry. HIF-1 伪, HIF-1 尾, VEGF,iNOS gene m RNA and protein expression were detected by RT-PCR and Western Blotting. Results: compared with the control group, the migration ability of hypoxia group increased (P0.05), apoptosis increased (P0.05), all were time-dependent. In hypoxia group, the expression of HIF-1 伪, HIF-1 尾, VEGF,iNOS gene mRNA and protein were all increased (P < 0. 05). Conclusion: hypoxia can enhance the migration and apoptosis of human skin microvascular endothelial cells, which may be related to the increased expression of HIF-1 伪 and its downstream genes.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R543.6

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