摘要
目的利用铁超载的方法在体外复制肝纤维化的氧应激模型,给予H2S供体硫氢化钠(NaHS)和内源性H2S作用的KATP通道阻滞剂格列苯脲(GLBN),论证氧应激下硫化氢对HSC细胞具有保护作用的假说。方法将HSC-T6分为8组:空白组、对照组、NaHS组(分为3个浓度组)、格列苯脲组(分为3个浓度组)。用CCK-8试剂盒测定HSC细胞增殖情况;用MDA试剂盒测定上清液MDA(丙二醛)含量;用SOD(超氧化物歧化酶)试剂盒测定上清液和裂解液中SOD活力;用RT-PCR试剂盒检测p38基因表达水平。结果1.Fe-NTA能够促进HSC的增殖,使p38基因表达水平降低、SOD活性降低、MDA浓度提高。对照组和空白组有明显差异(P<0.05);2.给予NaHS处理后,细胞增殖减慢,上清液中MDA含量下降,上清液、裂解液中SOD活性提高,p38基因表达水平增高(P<0.05);3.给予GLBN处理后,各项指标结果与给予NaHS处理后结果相对应,各组与对照组比较均有明显差异(P<0.05)。结论氧应激下硫化氢对HSC细胞具有保护作用,可抑制肝纤维化的发生发展。
Objective In order to investigate the changes and role of H2S in hepatic fibrosis, we make a oxidative stress model by incubating rat hepatic stellate cells with ferric nitri -lotriacetic acid( Fe -NTa) and analyzing the effects of Fe - NTA, sodium hydrosulfide ( NariS, H2S donor) and GLBN ( a inhibitor of KATP channel through which H2S reacts). Methods HSC - T6 were divided into eight groups: Blank group, Control group, Naris groups( to separate three groups) ;GLBN groups( to separate three groups). The MDA contents in culture supernatant and the SOD activity in culture supernatant and Cell Lysis Solution were detected. The HSC growth curve was detected by CCK - 8 and the expression of p38 was detected by RT - PCR. Results 1. In comparison with that in the blank group, the proliferations of hepatic stellate cells with Fe - NTA interference were remarkably increased, SOD activity reduced. P38 expression was down regulated, MDA contents were increased significantly between control group and blank group (P 〈 0.05). 2. After HSC treated with NariS, the proliferation of HSC reduced, MDA contents reduced, SOD activity increased,p38 expressions increased , After HSC treated with GLBN, the effects are opposite to the effects in groups which HSC treated with NariS. All the changes are significant as compared with that in the control group ( P 〈 0.05). Conclusion H2 S may be an antioxidant, which can protect liver and inhibit the development of Hepatic fibrosis.
出处
《青海医学院学报》
CAS
2008年第4期234-239,共6页
Journal of Qinghai Medical College
基金
青海省卫生厅2007年医药卫生科研指导性计划课题(2008-QY-02)
关键词
肝纤维化
肝星状细胞
硫化氢
硫氢化钠
Hepatic fibrosis Hepatic stellate cells Hydrogen sulfide Sodium hydrosulfide