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硫化氢对大鼠心脏成纤维细胞增殖的抑制作用

刘俊, 郝丹丹, 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

本文旨在研究硫化氢(hydrogen sulfide, H2S)对大鼠心脏成纤维细胞增殖的抑制作用。以原代培养新生大鼠心脏成纤维细胞(neonatal rat cardiac fibroblasts, NRCFs)为研究对象，用不同浓度血管紧张素II (angiotensin II, Ang II)或胎牛血清(fetal bovine serum, FBS)刺激NRCFs，建立NRCFs增殖模型。不同浓度硫氢化钠(NaHS，H2S的供体)处理该NRCFs增殖模型后，采用5’-溴-2’-脱氧尿嘧啶(5’-bromo-2’-deoxyuridine, BrdU)掺入法检测NRCFs增殖情况，用2’,7’-二氯荧光素乙酰乙酸盐(2’,7’-dichlorofluorescein diacetate, DCFH-DA)荧光探针法检测细胞活性氧类(reactive oxygen species, ROS)水平。结果显示，较低浓度的NaHS (1 × 10−5 mol/L)能促进FBS (2%、10%)对NRCFs的诱导增殖作用，但对Ang II (1 × 10−7 mol/L)所引起的NRCFs增殖的作用不明显，而较高浓度NaHS (5 × 10−5、1 × 10−4 mol/L)则显著抑制10% FBS和Ang II (1 × 10−7 mol/L)对NRCFs的诱导增殖作用；1 × 10−4 mol/L NaHS可显著降低Ang II (1 × 10−7 mol/L)诱导升高的细胞内ROS水平。以上结果表明，H2S在一定浓度范围内具有抑制NRCFs增殖的作用，其机制可能与H2S降低细胞内ROS有关。

关键词： 硫化氢; 大鼠; 心脏成纤维细胞; 血管紧张素II

分类号：R331

[Inhibitory effect of hydrogen sulfide on cardiac fibroblast proliferation.] [Article in Chinese]

LIU Jun, HAO Dan-Dan, ZHU Yi-Chun^*

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

Abstract

The aim of the present study was to investigate the role of hydrogen sulfide (H2S) in the proliferation of neonatal rat cardiac fibroblasts (NRCFs). Proliferation of NRCFs was induced by the presence of fetal bovine serum (FBS) or angiotensin II (Ang II) at various concentrations. The concentration-dependent effect of NaHS (donor of H2S) on NRCFs proliferation was examined. NRCFs proliferation was assessed by 5’-bromo-2’-deoxyuridine (BrdU) incorporation method. Reactive oxygen species (ROS) level was measured using the dye probe, 2’, 7’-dichlorofluorescein diacetate (DCFH-DA). The results showed that FBS- or Ang II-induced NRCFs proliferations were inhibited with the treatment of relatively high concentrations of NaHS (5 × 10−5 mol/L, 1 × 10−4 mol/L), but FBS-induced proliferation was increased by low concentration of NaHS (1 × 10−5 mol/L). Two or 6 h of Ang II (1 × 10−7 mol/L) treatment caused an increase of ROS level in NRCFs, while this increase was inhibited with NaHS (1 × 10−4 mol/L) treatment. These results suggest that H2S is an inhibitor of cardiac fibroblast at a certain concentration range. This inhibitory effect may be mediated by a reduction in intracellular ROS production.

Key words: hydrogen sulfide; rat; cardiac fibroblasts; angiotensin II

收稿日期：2011-05-15　　录用日期：2011-07-14

通讯作者：朱依纯　　E-mail: yczhu@shmu.edu.cn

引用本文：

刘俊, 郝丹丹, 朱依纯. 硫化氢对大鼠心脏成纤维细胞增殖的抑制作用[J]. 生理学报 2011; 63 (4): 353-358.

LIU Jun, HAO Dan-Dan, ZHU Yi-Chun. [Inhibitory effect of hydrogen sulfide on cardiac fibroblast proliferation.] [Article in Chinese]. Acta Physiol Sin 2011; 63 (4): 353-358 (in Chinese with English abstract).