硫化氢在SB203580影响肝星状细胞增殖、凋亡中的作用

【摘要】目的 探讨H₂S 在 P38 丝裂原活化蛋白激酶（P38MAPK）阻断剂 SB203580 影响大鼠肝星状细胞（HSC）-T6增殖、凋亡中的作用及对Ⅰ、Ⅲ型胶原蛋白mRNA表达的影响。方法设对照组（HSC加含10%胎牛血清的DMEM培养液）、二甲基亚砜（DMSO）组（对照组基础上加DMSO）、NaHS组（对照组基础上加NaHS至最适浓度）、SB组（DMSO组基础上加SB203580至最适浓度）、SB+NaHS组。采用四甲基偶氮唑盐（MTT）法测定SB203580及H2S对HSC的增殖抑制率（IR）；Annexin V-FITC/PI双染法流式细胞术（FCM）检测HSC凋亡率；逆转录PCR法检测Ⅰ、Ⅲ型胶原蛋白mRNA的表达。结果SB组（7.64±2.46）、SB+NaHS组（12.71±2.73）细胞凋亡率高于对照组（1.70±0.88），且SB+NaHS组高于SB组（均P＜0.05）；DMSO组（2.07±1.18）、NaHS组（2.56±1.23）与对照组差异无统计学意义。Ⅰ、Ⅲ型胶原蛋白mRNA在各组细胞中均表达，其中NaHS组表达高于对照组（P＜0.05），且表达量最多，条带最亮、最宽，SB组与SB+NaHS组表达低于对照组和NaHS组（P＜0.05），表达量较少，条带较暗。结论H₂S激活P38MAPK信号通路，且SB203580特异性阻断P38MAPK信号通路后，H2S刺激的HSC增殖受到抑制，凋亡得到促进。

The Role of Hydrogen Sulfide in the Effect of SB203580on Proliferation and Apoptosis of Hepatic Stellate Cells

Abstract]Objective To studythe role of hydrogen sulfide (H2S) in the effect of SB203580on proliferation andapoptosis of hepatic stellate cells and the effects of H2S on expressions of collagen Ⅰand collagen Ⅲ mRNA in hepatic stel-late cells.MethodsThere were five groups of HSC-T6cells in this study includingcontrol group (DMEM medium containing10% fetal bovine serum), dimethyl sulfoxide (DMSO) group, sodium hydrosulfide (NaHS）group，SB203580(SB）group and SB + NaHS group. MTT method was used to detect the cell proliferation and inhibition rate of HSC- T6cells treated by SB203580and H2S. The apoptotic rate of HSC- T6cells was detected by flow cytometry with annexin V- FITC/PI double staining. RT-PCR was used to detect the expressions of collagen Ⅰand collagen Ⅲ mRNA in HSC-T6.Results The apoptotic rate of HSC-T6cells was significantly higher in SB group and SB+NaHS group than that of control group, and the rate was significantly higher in SB+NaHS group than that of SB group (P<0.05). There was no significant difference in the apoptotic rate of HSC-T6cells between DMSO and NaHS groups than that of control group. The expressions of collagenⅠand collagen Ⅲ mRNA were found in five groups of cells. There was a higher expression of collagen Ⅰand collagen Ⅲ mRNA in NaHS group than that of control group (P<0.05). The expressions of collagen Ⅰand collagen Ⅲ mRNA were significantly lower in SB group and SB+NaHS group than those of control group and NaHS group (P<0.05).ConclusionH2S activated P38MAPK signal pathway. And P38MAPK was specifically blocked by SB203580in HSC-T6cells, which inhibited the cell proliferation stimulated by H2S and promoted the apoptosis.