硫化氢对氧应激大鼠肝星形细胞的影响

EFFECTS OF HYDROGEN SULFIDE ON RAT HEPATIC STELLATE CELLS IN OXIDATIVE STRESS

目的探讨硫化氢(H2S)对大鼠肝星形细胞(HSC)增殖和氧应激的影响。方法用铁超载的方法复制肝纤维化的氧应激模型,给予外源性H2S供体硫氢化钠(NaHS)和炔丙基甘氨酸(PPG,H2S合成关键酶胱硫醚-γ-裂解酶CSE抑制剂),通过绘制生长曲线法和用CCK-8细胞计数法测定不同浓度H2S和PPG分别作用于500μmol/L次氮基三乙酸铁(FeNTA)培养下的HSC6、12、24 h细胞上清液和裂解液中H2S浓度、超氧化物歧化酶(SOD)活力和丙二醛(MDA)含量,通过检测HSC的增殖情况、CSEmRNA表达水平、H2S浓度和氧化抗氧化指标(MDA、SOD)来探讨H2S对HSC氧应激的影响。结果FeNTA所致的增殖HSC氧应激损伤明显增加,抗氧化能力减弱。同时,CSEmRNA表达水平低下,H2S产生减少。给予PPG后,HSC增殖显著且CSEmRNA表达受到抑制。内源性H2S随着严重的氧应激损伤进一步减少。给予NaHS后,HSC增殖减少,CSEmRNA表达和H2S产生显著增加以缓解氧应激损害和增强抗氧化能力。结论H2S对HSC增殖具有一定的抑制作用,对FeNTA诱导的氧应激具有保护作用。

Objective To explore the effects of hydrogen sulfide on proliferation and oxidative stress of hepatic stellate cells HSC in rats. Methods Iron -overloaded method was used to establish the liver fibrosis model characterized by oxidative stress. After the NariS（ H2S donor）and the propargylglycine （PPG,a metabolic enzyme inhibitor of H2S） were given, the growth curves was drawn and CCK- 8 was measured by cell counting to reveal the effects of various concentrations of H2S and PPG on proliferation of HSC functioned with ferric nitrilotriacetic acid （FeNTA） （500 μmol/L）. After being given H2S and PPG, the concentrations of HES in cell lysate supematant and cell lysis solution, the activity of superoxide dismutase （SOD） and the contents of maleic dialdehyde （MDA）were detected after six hours, twelve hours and twenty - four hours respectively. In order to observe the effects of hydrogen sulfide on oxidative stress, the effects of FeNTA on the proliferation of HSC, the expression of CSEmRNA,MDA contents and SOD activity were detected respectively after giving FeNTA, NariS and PPG. Results FeNTA induced Oxidation damage of proliferated HSC was ncreased significantly with weakened anti - oxidant ability, meanwhile, the CSE mRNA expression were lower and the production of H2S decreased. After being given PPG, the proliferation of HSC accelerated and the CSE mRNA expression were restrained. At the same time, endogenous HiS decreased further with severe oxidation damage. After being given NariS, the proliferation of HSC decelerated and the CSE mRNA expression and the production of H2S increased markedly to alleviate oxidation damage and strengthen the anti - oxidant ability. Conclusion H2S has inhibitory effect on proliferation of HSC and protective effect on oxidative stress induced by FeNTA.