硫化氢对氧应激下大鼠肝星状细胞p38MAPK信号的影响

目的探讨硫化氢(H2S)对氧应激下大鼠肝星状细胞(HSC-T6)p38 MAPK信号的影响。方法对大鼠HSC-T6进行细胞培养,应用铁超载方法制备肝纤维化的氧应激模型,给予不同剂量的外源性H2S供体硫氢化钠(NaHS)和KATP通道抑制剂格列本脲(GLBN),作用于HSC-T6。在24、48h后分别提取培养后的HSC细胞,用RT-PCR方法测定p38 mRNA,Western-blot方法测定p38蛋白质以及磷酸化p38(p-p38)蛋白质表达水平。结果在应用Fe-NTA作用的大鼠HSC体外培养氧应激模型下,给予FeNTA24h和48h时后,模型组p38 mRNA和蛋白表达水平与空白组比较均明显升高,差异有统计学意义(P<0.05)。给予NaHS处理后,100、200和400μmol.L-1NaHS组的p38 mRNA和蛋白水平和模型组比较,表达逐渐降低,差异有统计学意义(P<0.05),并且组间差异均有统计学意义(P<0.05)。加入格列本脲后,p38mRNA和蛋白质表达水平均有所上升,但低于模型组。结论硫化氢通过抑制p38 MAPK而影响氧应激下大鼠HSC-T6。

The Impact of Hydrogen Sulfide on p38 MAPK Signal of the Rat Hepatic Stellate Cells under the Stress of Oxygen

Objective To explore the impact of hydrogen sulfide(H2S) on p38 MAPK signal of the rat hepatic stellate cells(HSC-T6) under the stress of oxygen.Methods After culturing the rat hepatic stellate cells(HSC-T6),the method of iron overload to prepare liver fibrosis oxygen stress model was applied Different doses of exogenous H2S donor sulfur hydrogenated sodium(NaHS) and KATP channel inhibitors glyburide(GLBN) were given to act on HSC-6 for 24h and 48h.After training,HSC cells were extracted respectively and p38 change process and its significance were determined with RT-PCR,Western-blot methods at different time points.Results After applying Fe-Nta at rats in vitro culture oxygen under stress HSC,compared with the blank group,the model group showed that after being given FeNTA for 24h and 48h,the expression of p38 mRNA and p-p38 protein increased significantly(P < 0.05).After being given NaHS,the level of p38 mRNA and p-p38 protein in the treatment group,compared with model group’s,had a significant decrease(P < 0.05).After glyburide being added,the expression of both p38 mRNA and p-p38 protein increased to a certain extent,however,still not as high as the level of the model group.Conclusion By means of restraining p38 MAPK signal path,H2S has an impact on the rat HSC-T6 under the stress of oxygen.