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Bone marrow mesenchymal stem cells modulate the expression of RhoA and P27 in hepatic stellate cells
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- Author:
- No author available
- Journal Title:
- WORLD CHINESE JOURNAL OF DIGESTOLOGY
- Issue:
- 32
- DOI:
- 10.3969/j.issn.1009-3079.2009.32.002
- Key Word:
- 骨髓间充质干细胞;肝星状细胞;RhoA;P27;细胞周期
Abstract： 目的:观察体外大鼠骨髓间充质干细胞(MSCs)对肝星状细胞(HSCs)RhoA信号因子及其细胞周期调控因子P27表达的影响,探讨MSCs调控HSCs细胞周期G1/S转换机制.方法:贴壁筛选法培养、纯化SD大鼠MSCs,传代至第4代使用;大鼠肝星状细胞(HSC-T6)系及纤维原细胞系冻融后传代使用.应用6孔塑料细胞培养盒,每孔使用半透膜(transwellinsert)建立上下双层细胞共培养体系,常规培养.实验分3组:空白对照组、阴性对照组、MSCs实验组.用WST-8法对HSCs增殖率进行测定:流式细胞仪检测细胞周期:RT-PCR、Western blot检测MSCs与HSCs共培养后HSCs内RhoA,P27 mRNA和蛋白的表达.结果:(1)HSCs与MSCs共培养24 h后,HSCs表现明显增殖抑制(P<0.01),且呈现时间依赖性,MSCs实验组与空白对照组、阴性对照组比较均有显著性差异(均P<0.01).(2)共培养12 h后,MSCs可阻滞HSCs由G0G1期向S期转换,使G0/G1期细胞增多,S期细胞减少,与空白对照组、阴性对照组比较差异均有统计学意义(P<0.01),(3)共培养12 h后,MSCs实验组RhoAmRNA表达与空白对照组、阴性对照组比较均有统计学差异(P<0.05,P<0.01),随时间的延长呈递减趋势;共培养后,MSCs实验组P27mRNA表达与空白对照组、阴性对照组比较均无统计学差异.(4)共培养12 h后,MSCs实验组RhoA蛋白表达与空白对照组、阴性对照组比较均有统计学差异(均P<0.01),随时间的延长呈递减趋势:共培养12 h后,MSCs实验组P27蛋白与空白对照组、阴性对照组比较均有统计学差异(均P<0.01),随时间的延长呈递增趋势.(5)相关性分析显示:RhoA与P27mRNA的表达无明显相关(r=0.105);RhoA与P27蛋白的表达呈显著负相关(r=-0.943,P<0.01).结论:MSCs抑制HSCs增殖的机制可能是通过RhoA-P27通路调控HSCs细胞周期改变;RhoA活性的下调可能是引起HSCs内P27蛋白表达增加的原因.
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