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低氧诱导因子1参与常氧条件下硫化氢促血管内皮细胞迁移的作用

陶蓓蓓, 张彩彩, 刘姝媛, 朱依纯^*

复旦大学上海医学院生理学与病理生理学系，上海 200032

摘要

本文旨在探讨常氧条件下硫化氢(H2S)促进血管内皮细胞迁移的分子机制。采用划痕迁移模型研究常氧条件下NaHS (H2S供体)对恒河猴视网膜-脉络膜内皮细胞系RF/6A细胞迁移的作用；用Real-time PCR方法检测RF/6A细胞内低氧诱导因子1α (hypoxia-inducible factor 1α, HIF-1α) mRNA水平；用Western blot检测RF/6A细胞内HIF-1α的蛋白水平；用2’,7’-二氯荧光素乙酰乙酸盐(2’,7’-dichlorofluorescein diacetate, DCFH-DA)荧光探针法检测RF/6A细胞内活性氧(reactive oxygen species, ROS)水平。结果显示，不同浓度(10~100 μmol/L) NaHS对RF/6A细胞有显著的促迁移作用，该作用可以被HIF-1抑制剂氯化镉(CdCl2, 50 µmol/L)所阻断。NaHS能够在常氧培养条件下呈剂量和时间依赖性地显著提高HIF-1α的蛋白水平，并且对HIF-1α mRNA水平有快速、持久的提高作用。另外，NaHS能够显著降低常氧条件下RF/6A细胞中的ROS水平。以上结果提示，HIF-1参与常氧条件下H2S促内皮细胞迁移的作用，而ROS作为HIF-1α的上游调节因子，可能也参与H2S促进内皮细胞迁移的过程。

关键词： 硫化氢; 内皮细胞; 低氧诱导因子1; 活性氧

分类号：R331

[Involvement of HIF-1 in the migration-promoting effects of hydrogen sulfide in vascular endothelial cells under normoxic conditions.] [Article in Chinese]

TAO Bei-Bei, ZHANG Cai-Cai, LIU Shu-Yuan, ZHU Yi-Chun^*

Department of Physiology and Pathophysiology, Fudan University Shanghai Medical College, Shanghai 200032, China

Abstract

The purpose of this study was to investigate the molecular mechanisms whereby hydrogen sulfide (H2S) exerts the promoting effect on vascular endothelial cells migration. We used wound healing assay to study the effect of NaHS (H2S donor) on the migration ability of rhesus retinal pigment epithelial cell line, RF/6A cells, under normoxic conditions. Real-time PCR was used to measure hypoxia-inducible factor 1α (HIF-1α) mRNA level. Western blot was used to measure the expression of HIF-1α protein. The probe 2’,7’-dichlorofluorescein diacetate (DCFH-DA) was used to measure intracellular reactive oxygen species (ROS) level. The results showed that NaHS (10–100 μmol/L) could significantly promote RF/6A cells migration under normoxic conditions, and this effect could be inhibited by 50 µmol/L HIF-1 inhibitor, CdCl2. NaHS increased the protein level of HIF-1α in a dose- and time-dependent manner, and up-regulated the mRNA level of HIF-1α quickly and continuously. Moreover, NaHS could significantly decrease ROS levels in RF/6A cells under normoxic conditions. These results suggest HIF-1 may mediate the promoting effect of H2S on vascular endothelial cells migration under normoxic conditions. ROS, as an upstream regulator of HIF-1α, may be involved in the migration-promoting effect of H2S.

Key words: hydrogen sulfide; endothelial cell; hypoxia-inducible factor 1; reactive oxygen species

收稿日期：2012-02-17　　录用日期：2012-04-01

通讯作者：朱依纯　　E-mail: yczhu@shmu.edu.cn

引用本文：

陶蓓蓓, 张彩彩, 刘姝媛, 朱依纯. 低氧诱导因子1参与常氧条件下硫化氢促血管内皮细胞迁移的作用[J]. 生理学报 2012; 64 (2): 129-134.

TAO Bei-Bei, ZHANG Cai-Cai, LIU Shu-Yuan, ZHU Yi-Chun. [Involvement of HIF-1 in the migration-promoting effects of hydrogen sulfide in vascular endothelial cells under normoxic conditions.] [Article in Chinese]. Acta Physiol Sin 2012; 64 (2): 129-134 (in Chinese with English abstract).