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热休克蛋白90在硫化氢抗化学性缺氧诱导心肌细胞氧化应激损伤中的作用 被引量:4

Roles of heat shock protein 90 in the blockage of H_2S against cardiomyocyte injuries induced by chemical hypoxia
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摘要 目的:探讨热休克蛋白90(HSP90)在硫化氢(H2S)对抗化学性低氧模拟剂氯化钴(CoCl2)诱导H9c2心肌细胞氧化应激损伤中的作用。方法:应用CoCl2处理H9c2心肌细胞,建立化学性缺氧损伤心肌细胞的实验模型。在CoCl2处理H9c2心肌细胞前30min,把硫氢化钠(NaHS,H2S的供体)加入培养基中,作为预处理。应用高效液相色谱法(HPLC)检测细胞内ATP的含量;罗丹明123(Rh123)染色荧光显微镜照相检测线粒体膜电位(MMP);超氧化物歧化酶(SOD)检测试剂盒检测SOD活性;免疫印迹法(Western blotting)检测血红素氧合酶-1(HO-1)的表达。结果:600μmol/LCoCl2明显地降低H9c2心肌细胞内SOD活性、ATP水平及MMP,并增加HO-1表达。400μmol/LNaHS预处理可显著地抑制CoCl2诱导的细胞毒性及氧化应激反应,使SOD活性、ATP水平及MMP提高,HO-1表达减少。热休克蛋白90抑制剂17-丙烯胺基-17去甲氧基格尔德霉素(17AAG)能明显地阻断H2S对CoCl2诱导的细胞毒性和氧化应激反应的抑制作用,使细胞内ATP水平及MMP降低,HO-1表达增多,但对SOD活性的影响不明显。结论:热休克蛋白90可通过抑制化学性缺氧引起的氧化应激反应来介导H2S的心肌保护作用。 AIM : To explore the roles of heat shock protein 90 (HSP90) in the blockage of hydrogen sulfide (H2S) against chemical hypoxiamimetic agent (cobalt chloride, CoCl2 ) -induced oxidative stress injuries in H9c2 cardiac cell. METHODS: H9c2 cells were treated with CoCl2 to set up the chemical hypoxiainduced the model of cardiomyocyte injury. Sodium hydrosulfide (NariS, a H2S donor) was added into medium for 30 min before CoCl2 treatment. ATP content was detected by high performance liquid chromatogram (HPLC). Mitochondrial membrane potential (MMP) was measured by rhodamine123 ( Rh123 ) staining and photofluorography. The activity of superoxide dismutase (SOD) was observed using a SOD kit. The expression of heine oxygenase - 1 ( HO - 1 ) was evaluated by Western blotting. RESUETS: CoCl2 at concentration of 600 μmol/L significantly reduced SOD activity, ATP level and MMP, and enhanced the expression of HO - 1 in H9c2 cells. Pretreatment with 400μmol/L NariS dramatically inhibited the cytotoxicity induced by CoCl2 , increased SOD activity, ATP level and MMP, decreased HO - 1 expression. 17 - allylamino - 17 demethoxygeldanamycine( 17AAG), an inhibitor of HSP90, obviously blocked the inhibitory effect of H2S on the COCl2 - induced cytotoxiciy, reduced the levels of ATP and MMP, increased HO - 1 expression. However, no significantly influence on SOD activity was observed. CONCLUSION: HSP90 may mediate the cardioprotection of H2 S via inhibiting the oxidative stress induced by chemical hypoxia.
出处 《中国病理生理杂志》 CAS CSCD 北大核心 2009年第12期2329-2333,共5页 Chinese Journal of Pathophysiology
基金 广东省科技计划资助项目(No.2008B080703053 No.2007B080701030)
关键词 热休克蛋白90 硫化氢 氯化钴 心肌细胞 氧化性应激 Heat-shock proteins 90 Hydrogen sulfide Cobalt chloride Cardiomyocytes Oxidative stress
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参考文献16

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共引文献44

同被引文献75

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