低氧增加人皮肤微血管内皮细胞系迁移并促进凋亡

Hypoxia promotes the migration and apoptosis of human dermal micro-vascular endothelial cells

目的研究低氧环境对人皮肤微血管内皮细胞(HDMEC)迁移、凋亡及相关因子HIF-1α、VEGF、i NOS基因及蛋白表达的影响。方法低氧培养人微血管内皮细胞,分为常氧组(对照组)和低氧组(Co Cl2模拟化学低氧)。CCK-8细胞生存实验确定合适处理浓度(200μmol/L Co Cl2),划痕实验检测细胞迁移能力,流式细胞技术观察细胞凋亡;用RT-PCR和Western blot技术检测HIF-1α、VEGF、i NOS mRNA和蛋白表达。结果与常氧组比较,低氧组细胞迁移能力增加(P<0.05),凋亡增多(P<0.05),均呈时间依赖性。低氧组HIF-1α、HIF-1β、VEGF、i NOS mRNA表达较常氧组比较均增加(P<0.05),HIF-1α、VEGF、i NOS蛋白表达较常氧组比较均增加(P<0.05),具有一定时间依赖性。结论低氧能增强人皮肤微血管内皮细胞迁移能力,并促进其细胞凋亡,其可能机制与HIF-1α、VEGF、i NOS蛋白表达升高有关。

Objective To study the effects of hypoxia on the cell migration,apoptosis and expression of related genes and proteins of human dermal microvascular endothelial cells（ HDMEC）. Methods To culture HDMEC in hypoxia condition by putting Co Cl2 in the cell incubator and all the subjects were divided to two groups which included normal group and hypoxia group. To find the appropriate Co Cl2 dose by CCK-8 experiment and the 200 μmol/L Co Cl2 was the best to establish hypoxia model. The migration of cells was measured by wound healing test. Apoptosis rate was detected by flow cytometry. The gene expressions were detected by RT-PCR and the protein levels were detected by Western blot. Results The migration ability of cells was enhanced in hypoxia condition（ P〈0. 05）. The apoptosis rate was increased in hypoxia condition（ P〈0. 05）. The gene expressions of HIF-1α,HIF-1β,VEGF,i NOS were increased comparing with normal groups depending on time（ P〈0. 05）. The protein expressions of HIF-1α,VEGF,i NOS were increased comparing with normal groups depending on time（ P 0. 05）. Conclusions Cell migration,apoptosis of HDMEC were influenced by hypoxia. It may be explained by increasing expression of related gene and proteins like HIF-1α,VEGF,i NOS in hypoxia condition.