CAJ | 학술논문

目的:通过调控心脏干细胞(CSCs)中的miRNA-146b的表达水平,探讨miRNA-146b对CSCs迁移、增殖和分化的影响.方法:取1～3d的C57BL/6小鼠的CSCs,经过磁珠分选,选取c-kit+ CSCs培养.c-kit+CSCs在骨髓间充质干细胞培养液刺激下,后以实时聚合酶链式反应(RealtimePCR)检测c-kit+ CSCs的miRNA146b表达水平.进一步将c-kit+ CSCs培养3d后分为5组进行转染:空白对照组、上调miRNA-146b组、上调阴性对照组、下调miRNA-146b组、下调阴性对照组.以CCK-8法检测转染后c-kit+ CSCs增殖,transwell法检测转染后c-kit+CSCs迁移.PCR法检测GATA-4、MEF2c、TNI、β-MHC等分化相关基因表达水平.结果:在骨髓间充质干细胞培养液刺激下c-kit+CSCs的miRNA-146b表达上调(P＜0.05).在miRNA-146b的前体或抑制剂转染后,能有效使c-kit+ CSCs细胞内miRNA-146b过表达或抑制,与上调阴性对照组比较,上调miRNA-146b组的miRNA-146b表达明显升高[(2.26±1.57)比(340.54±36.63)],与下调阴性对照组比较,下调miRNA-146b组miRNA-146b表达明显下降[(1.11±0.16)比(0.66±0.04)],P均＜0.01.同阴性对照组比较,上调miRNA-146b组c-kit+CSC迁移能力明显降低[(70.6±9.1)比(47±5.3)],下调miRNA-146b组的c-kit+ CSCs迁移能力明显增强[(39.5±4.1)比(48.0±8.8)],P均＜0.05.与上调阴性对照组比较,上调miRNA-146b组的c-kit+ CSCs增殖活性显著降低；与下调阴性对照组比较,下调miRNA-146b组的c-kit+CSCs增殖活性显著增强,P均＜0.05.miRNA-146b对分化无明显影响.结论:miRNA-146b能够抑制c-kit+ CSCs增殖和迁移能力.
목적:통과조공심장간세포(CSCs)중적miRNA-146b적표체수평,탐토miRNA-146b대CSCs천이、증식화분화적영향.방법:취1～3d적C57BL/6소서적CSCs,경과자주분선,선취c-kit+ CSCs배양.c-kit+CSCs재골수간충질간세포배양액자격하,후이실시취합매련식반응(RealtimePCR)검측c-kit+ CSCs적miRNA146b표체수평.진일보장c-kit+ CSCs배양3d후분위5조진행전염:공백대조조、상조miRNA-146b조、상조음성대조조、하조miRNA-146b조、하조음성대조조.이CCK-8법검측전염후c-kit+ CSCs증식,transwell법검측전염후c-kit+CSCs천이.PCR법검측GATA-4、MEF2c、TNI、β-MHC등분화상관기인표체수평.결과:재골수간충질간세포배양액자격하c-kit+CSCs적miRNA-146b표체상조(P＜0.05).재miRNA-146b적전체혹억제제전염후,능유효사c-kit+ CSCs세포내miRNA-146b과표체혹억제,여상조음성대조조비교,상조miRNA-146b조적miRNA-146b표체명현승고[(2.26±1.57)비(340.54±36.63)],여하조음성대조조비교,하조miRNA-146b조miRNA-146b표체명현하강[(1.11±0.16)비(0.66±0.04)],P균＜0.01.동음성대조조비교,상조miRNA-146b조c-kit+CSC천이능력명현강저[(70.6±9.1)비(47±5.3)],하조miRNA-146b조적c-kit+ CSCs천이능력명현증강[(39.5±4.1)비(48.0±8.8)],P균＜0.05.여상조음성대조조비교,상조miRNA-146b조적c-kit+ CSCs증식활성현저강저；여하조음성대조조비교,하조miRNA-146b조적c-kit+CSCs증식활성현저증강,P균＜0.05.miRNA-146b대분화무명현영향.결론:miRNA-146b능구억제c-kit+ CSCs증식화천이능력.