Department of Pharmacology, School of Pharmacy and Institute of Biomedical Sciences, Fudan University, Shanghai, China.
Biol Pharm Bull. 2010;33(9):1550-4. doi: 10.1248/bpb.33.1550.
Hydrogen sulfide (H2S) is known to have pro-angiogenic properties in mammals. In this study, we examined H2S played the role in pro-angiogenesis mediated by hypoxia-inducible factor (HIF)-1alpha under hypoxic conditions. Rat brain capillary endothelial cells (ECs) were treated with NaHS (a H2S donor) pretreated vascular smooth muscle cells (VSMCs) conditioned media. ECs proliferation was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. ECs migration was assessed by chemotaxis chamber assay. Angiogenesis-associated gene expression levels were determined by reverse transcription-polymerase chain reaction (RT-PCR). HIF-1alpha and vascular endothelial growth factor (VEGF) accumulation was analyzed by Western blotting. HIF-1 binding activity was measured by electrophoretic mobility shift assay (EMSA). We found H2S induced both endothelial proliferation and migration in mimic hypoxic condition. In addition, H2S promoted VEGF and HIF-1alpha mRNA levels. H2S also significantly upregulated HIF-1alpha and VEGF protein levels and increased HIF-1alpha binding activity under hypoxic condition. Our findings suggest that HIF-1/VEGF is involved in H2S promotes proliferation and migration of ECs.
硫化氢 (H2S) 已知在哺乳动物中具有促血管生成特性。在这项研究中，我们研究了在缺氧条件下，HIF-1α 介导的缺氧诱导因子 (HIF-1α) 下 H2S 发挥的作用。用 NaHS（H2S 供体）预处理血管平滑肌细胞（VSMCs）条件培养基处理大鼠脑微血管内皮细胞（ECs）。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐 (MTT) 测定法评估 ECs 增殖。通过趋化性室测定评估 ECs 迁移。通过逆转录-聚合酶链反应 (RT-PCR) 测定血管生成相关基因表达水平。通过 Western blot 分析 HIF-1α 和血管内皮生长因子 (VEGF) 积累。通过电泳迁移率变动分析 (EMSA) 测量 HIF-1 结合活性。我们发现 H2S 在模拟缺氧条件下诱导内皮细胞增殖和迁移。此外，H2S 促进了 VEGF 和 HIF-1α mRNA 水平。H2S 还显著上调了 HIF-1α 和 VEGF 蛋白水平，并在缺氧条件下增加了 HIF-1α 结合活性。我们的研究结果表明，HIF-1/VEGF 参与了 H2S 促进 ECs 增殖和迁移的过程。
