>
网站首页期刊介绍通知公告编 委 会投稿须知电子期刊广告合作联系我们
最新消息:
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
作者:黄若兰1  张忠2  陈铭泰3  王玲1  常晓1  乔秋杰1 
单位:1. 深圳市中医院 重症医学科, 广东 深圳 518033;
2. 深圳市中医院 心血管科, 广东 深圳 518033;
3. 广州中医药大学, 广东 广州 518000
关键词:骨髓间充质干细胞 膜微粒 血管平滑肌细胞 迁移 凋亡 
分类号:R329.2
出版年·卷·期(页码):2016·35·第四期(475-481)
摘要:

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。

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.

参考文献:

[1] 王凤,章怡祎,刘萍.动脉粥样硬化动物模型的研究进展[J].东南大学学报:医学版,2014,33(2):235-238.
[2] 刘晶,马坤岭,倪杰,等.动脉粥样硬化模型小鼠血管平滑肌细胞原代培养及鉴定[J].东南大学学报:医学版,2011,30(4):537-540.
[3] MARX S O,TOTARY-JAIN H,MARKS A R.Vascular smooth muscle cell proliferation in restenosis[J].Circ Cardiovasc Interv,2011,4(3):104-111.
[4] 吴校林,江洪,陈静,等.Kindlin-2对血管平滑肌细胞迁移和黏附的影响及其机制的实验研究[J].中华心血管病杂志,2014,42(11):938-943.
[5] 金杰,廖明芳,王亮,等.微小RNA调节血管平滑肌细胞增殖及其参与心血管疾病病理形成的研究进展[J].中华心血管病杂志,2015,43(9):837-840.
[6] 蒋珽,项阳.MicroRNA在冠心病中的作用研究[J].东南大学学报:医学版,2012,31(5):639-643.
[7] 黎叶飞,盛祖龙,姚玉宇,等.两种骨髓间充质干细胞移植途径治疗急性心肌梗死的比较研究[J].东南大学学报:医学版,2011,30(5):687-691.
[8] CHEN L,WANG Y,PAN Y,et al.Cardiac progenitor-derived exosomes protect ischemiac myocardium from acute ischemia/reperfusion injury[J].BBRC,2013,431(4):566-571.
[9] van BERLO J H,KANISICAK O,MAILLET M,et al.C-kit+cells minimally contribute cardiomyocytes to the heart[J].Nature,2014,509(3):337-345.
[10] 王艳,石蓓.微小核糖核酸与干细胞移植治疗心肌梗死的研究进展[J].中国循环杂志,2015,30(207):916-918.
[11] ELLISON G M,VICINANZA C,SMITH A J,et al.Adult c-kitpos cardiac stem cells are necessary and sufficient for functional cardiac regeneration and repair[J].Cell,2013,8(3):827-842.
[12] 吴原波,尹琪,金培峰,等.SDF-1α在体内对骨髓间充质干细胞的保护作用及其对缺血心脏修复的影响[J].温州医学院学报,2011,41(2):136-140.
[13] 耿志敏,王珏,范鸿洋,等.骨髓间充质干细胞膜微粒促进大鼠心肌梗死后血管新生[J].中国病理生理杂志,2015,8(3):1371-1375.
[14] NIEUWLAND R,STURK A.Why do cells release vesicles[J].Thromb Res,2010,125(1):49-51.
[15] MAYERS J R,AUDHYA A.Vesicle formation within endosomes:an ESCRT marks the spot[J].Commun Integr Biol,2012,5(1):50-56.
[16] HEINRICH E M,DIMMELER S.MicroRNAs and stem cells:control of pluripotency,reprogramming,and lineage commitment[J].Circ Res,2012,110(3):1014-1022.
[17] LEONARDO T R,SCHULTHEISZ H L,LORING J F,et al.The functions of microRNAs in pluripotency and reprogramming[J].Nat Cell Biol,2012,14(3):1114-1121.
[18] COLLINO F,DEREGIBUS M C,BRUNO S,et al.Microvesicles derived from adult human bone marrow and tissue specific mesenchymal stem cells shuttle selected pattern of miRNAs[J].PLoS One,2010,5(7):11803-11803.
[19] 龙仙萍,崔璨,陈攀科,等.降钙素基因相关肽和受体活性修饰蛋白1对血管平滑肌细胞迁移的影响及机制[J].中华心血管病杂志,2015,43(6):537-541.
[20] LI T S,TAKAHASHI M,OHSHIMA M,et al.Myocardial repair achieved by the intramyocardial implantation of adult cardiomyocytes in combination with bone marrow cells[J].Cell Transplant,2008,17(6):695-703.
[21] GATTI S,BRUNO S,DEREGIBUS M C,et al.Microvesicles derived from human adult mesenchymal stem cells protect against ischaemia-reperfusion-induced acute and chronic kidney injury[J].Nephrol Dial Transplant,2011,26(5):1474-1484.

服务与反馈:
文章下载】【发表评论】【查看评论】【加入收藏
提示:您还未登录,请登录!点此登录
您是第 405844 位访问者


copyright ©《东南大学学报(医学版)》编辑部
联系电话:025-83272481 83272483
电子邮件:
bjb@pub.seu.edu.cn

苏ICP备09058364