微小核糖核酸-497调控骨髓间充质干细胞向心肌样细胞分化的作用及其机制研究  

Regulation and mechanism of miR-497 on differentiation of bone marrow mesenchymal stem cells into cardiomyocytes

作　　者：吴坚 徐杨超 张明[2] WU Jian;XU Yang-chao;ZHANG Ming(Department of Basic Medicine,Jiangxi Medical College,Shangrao 334000,Jiangxi Province,China;Department of Cardiology,Jiangxi People’s Hospital,Nanchang 330000,Jiangxi Province,China)

机构地区：[1]江西医学高等专科学校基础医学部,江西上饶334000 [2]江西省人民医院心血管内科,江西南昌330000

出　　处：《中国临床药理学杂志》2020年第2期129-132,共4页The Chinese Journal of Clinical Pharmacology

基　　金：江西省卫生计生委科技计划基金资助项目(20185015)。

摘　　要：目的探讨微小核糖核酸-497(miR-497)在小鼠骨髓间充质干细胞(BMSCs)向心肌样细胞分化中的作用及机制。方法将BMSCs分为4组:空白组(未作任何处理)、阳性对照组(5-氮杂胞嘧啶8μmol·L-1体外诱导24 h)、阴性对照组(转染mimics-NC序列)和miR-497 mimics组(转染miR-497 mimics序列)。以蛋白质印迹法检测Notch1、Delta样配体1(DII-1)蛋白表达水平(灰度值),以AnnexinV-FTTC/PI双染法检测BMSCs凋亡率。结果空白组、阳性对照组、阴性对照组和miR-497mimics转染组的Notch1蛋白表达量分别为1.34±0.27,0.75±0.20,1.28±0.29和0.62±0.17;这4组的DII-1蛋白表达量分别为0.67±0.15,0.44±0.13,0.62±0.15和0.32±0.09,阳性对照组和miR-497mimics组与空白组比较,Notch1、DII-1蛋白表达量均明显降低,差异均有统计学意义(均P<0.05);miR-497mimics组与阳性对照组和阴性对照组比较,这2种蛋白的差异均有统计学意义(均P<0.05)。在转染72 h,上述4组BMSCs凋亡率分别为(24.86±5.77)%,(56.93±15.82)%,(28.47±8.36)%和(33.59±8.25)%,阳性对照组、阴性对照组和miR-497mimics组与空白组比较,凋亡率均明显升高,差异均有统计学意义(均P<0.05);miR-497mimics组与阳性对照组比较,凋亡率的差异有统计学意义(P<0.05)。结论上调BMSCs中miR-497的表达,可通过抑制Notch信号通路,激活诱导骨髓间充质干细胞向心肌样细胞分化,且诱导分化能力和安全性均优于5-氮杂胞嘧啶。Objective To discuss the effect of miR-497 on the differentiation of bone marrow-derived mesenchymal stem cells(BMSCs)into cardiomyocytes by inhibition of Notch pathway.Methods The BMSCs were divided into four groups:normal group(without any treatment),positive control group(treated with 8μmol·L-15-Aza-2’-deoxycytidine for 48 h),negative control group(treated with mimics-NC for 48 h)and miR-497 mimics group(treated with miR-497 mimics for 48 h).The expression levels(grey value)of Notch1,Delta-likeligand 1(DII-1)were detected by Western blot.The apoptosis rate of BMSCs was detected by Flow cytometry.Results The expression levels of Notch1 protein in normal group,positive control group,negative control group and miR-497 mimics group were 1.34±0.27,0.75±0.20,1.28±0.29,0.62±0.17;the expression levels of Dll1 protein in the four groups were 0.67±0.15,0.44±0.13,0.62±0.15,0.32±0.09.Comparison between positive control group,miR-497 mimics group and normal group,the difference of the factors were significantly(all P<0.05);comparison between miR-497 mimics group with positive control group and negative control group,the difference of the factors were significantly(P<0.05).Seventy-two hours after transfection,the apoptosis rates of BMSCs in the four groups were(24.86±5.77)%,(56.93±15.82)%,(28.47±8.36)%,(33.59±8.25)%.Comparison between positive control group,negative control group and miR-497 mimics group with normal group,the difference was significantly(all P<0.05);comparison between miR-497 mimics group and positive control group,the difference was significantly(P<0.05).Conclusion MiR-497 in BMSCs has the potential to effectively induce the differentiation of BMSCs into cardiomyocytes via inhibition of Notch signal pathway.

关 键 词：骨髓间充质干细胞 微小核糖核酸-497 心肌样细胞分化 5-氮杂胞嘧啶 NOTCH通路 

分 类 号：R97[医药卫生—药品]