天津医药

• 实验研究 •    下一篇

硫化氢在SB203580影响肝星状细胞增殖、凋亡中的作用

徐霞1,李睿2,任嫱1,刘芳2,宋丽秀2,3,郑勇2,3,陈卫刚2   

  1. 1. 新疆石河子大学医学院省部共建教育部重点实验室
    2. 新疆石河子大学第一附属医院消化内科
    3.
  • 收稿日期:2013-05-03 修回日期:2013-07-04 出版日期:2013-11-15 发布日期:2013-11-15
  • 通讯作者: 徐霞

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

XU Xia 1,LI Rui 2,REN Qiang 1,LIU Fang 2,SONG Li xiu2,ZHENG Yong 2,CHEN Wei gang2   

  • Received:2013-05-03 Revised:2013-07-04 Published:2013-11-15 Online:2013-11-15
  • Contact: XU Xia

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摘要：

【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。 

关键词: 硫化氢, 细胞增殖, 细胞凋亡, p38丝裂原活化蛋白激酶类, 肝星状细胞, SB203580

Abstract:

[Abstract]   Objective   To studythe role of hydrogen sulfide (H2S) in the effect of SB203580on proliferation and
apoptosis of hepatic stellate cells and the effects of H2S on expressions of collagen Ⅰand collagen Ⅲ mRNA in hepatic stel-late cells.  Methods   There 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).   Conclusion     H2S 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.

Key words: hydrogen sulfide, cell proliferation, apoptosis, p38mitogen-activated protein kinases, hepatic stellate cell, SB203580