miRNA145介导骨髓间充质干细胞膜微粒减少血管平滑肌细胞迁移

摘要

Objective: To investigate the impact and mechanism of microparticles derived from bone marrow mesenchymal stem cells ( MSC-MPs ) on migration and apoptosis of vascular smooth muscle cells ( VSMCs ) . Methods:The bone marrow mesenchymal stem cells were induced to release microparticles ( MSC-MPs ) through apoptosis for the experiment.Five experimental groups were listed as follows: control group ( A group );microparticles group ( B group ); miRNA145 group ( C group ); microparticles +antago-miRNA145 group ( D group); microparticles +miRNA145 group ( E group ) .Transwell assay was applied to detect the migration of smooth muscle cells, Western blot was used to determine the expression of protein, and flow cytometry and MTT were employed to examine the level of apoptosis in each group.The expression of miRNA145 was determined by real-time polymerase chain reaction( RT-PCR) .Results:The number of VSMC migration was significantly lower in E group(40.4 ±3.0) than that in D group (69.0 ±5.6)(P<0.05).The MTT and flow cytometry showed that the expression of miRNA145 , VSMC proliferation inhibition rate and apoptosis rate were highest in E group and lowest in D group(P<0.05).The result of caspase-3 expression evaluated by Western blot showed the same tendency. Conclusion:MSC-MPs could reduce the apoptosis of VSMC and inhibit the migration of VSMC, while miRNA145 is involved in the process.%目的：探讨骨髓间充质干细胞（ MSC）膜微粒对血管平滑肌细胞（ VSMC）增殖迁移的影响及其机制。方法：诱导 MSC 凋亡释放膜微粒（ MSC－MPs ）用于实验，实验分为对照组（ A 组）、膜微粒组（ B 组）、miRNA145组（ C组）、膜微粒加抗miRNA145组（ D组）及膜微粒加miRNA145组（ E组）5组。细胞迁移实验评价VSMC的迁移能力，免疫印迹法检测蛋白表达水平，流式细胞术、四甲基偶氮唑盐比色法（ MTT法）等比较各组VSMC凋亡水平。以实时定量聚合酶链反应检测miRNA145表达水平。结果：E组VSMC迁移数量最少[（40．4±3．0）个]，D组最多[（69．0±5．6）个]，差异具有统计学意义（P＜0．05）；miRNA145的表达水平E组最高，A组与D组最低（ P＜0．05）；MTT结果提示E组VSMC增殖抑制率及凋亡率最高，D组最低（ P＜0．05）；半胱氨酸蛋白酶－3免疫印迹法结果也类似。结论：MSC－MPs可减少VSMC凋亡，抑制VSMC迁移，而miRNA145参与了这一过程。