摘要
目的探讨微小RNA(micro RNA,miR)-19b对骨髓间充质干细胞(bone marrow mesenchymal stem sells,BMSCs)分化的影响,并研究其协同5-氮胞苷(5-azacytidine,5-aza)对BMSCs分化成心肌样细胞的作用。方法分离培养SD大鼠BMSCs,细胞鉴定后使用miR-19b mimics/inhibitor转染BMSCs,实时荧光定量聚合酶链反应(real time quantitative polymerase chain reaction,qRT-PCR)检测转染效率。采用10μmol/L 5-aza干预转染后的BMSCs,观察细胞形态学变化。采用Western blot检测诱导后BMSCs中心房钠尿肽(atrial natriuretic peptide,ANP)、心肌肌钙蛋白I(cardiac troponin I,cTnI)、和心肌肌钙蛋白T(cardiac troponin T,cTnT)表达。结果与5-aza诱导的BMSCs相比,miR-19b mimics转染后的BMSCs无明显形态学变化,5-aza诱导的miR-19b mimics组细胞形态变化最大,细胞体积变大,细胞聚集呈涡旋样,而5-aza诱导的miR-19b inhibitor组细胞形态变化不明显。与对照组比较,miR-19b mimics组ANP、cTnI及cTnT蛋白表达水平明显升高,而miR-19b inhibitor组ANP、cTnI及cTnT蛋白表达水平明显降低。结论miR-19b无诱导BMSCs分化的能力,但协同5-aza诱导BMSCs可提高其向心肌样细胞分化的能力。
Objectives To investigate the effect of micro RNA(miR)-19b on the differentiation of bone marrow mesenchymal stem sells(BMSCs)and its synergistic effect with 5-azacytidine(5-aza)on the differentiation of BMSCs into cardiomyocyte-like cells.Methods BMSCs of SD rats were isolated and cultured.After cell identification,BMSCs were transfected with miR-19b mimics/inhibitor.The transfection efficiency was detected by real time quantitative polymerase chain reaction(qRT-PCR).10μmol/L 5-aza was used to intervene the transfected BMSCs.The morphological changes of BMSCs were observed.Western blot was used to detect the expression of atrial natriuretic peptide(APN),cardiac troponin I(cTnI)and cardiac troponin T(cTnT)in BMSCs after induction.Results Compared with BMSCs induced by 5-aza,there was no significant morphological change in BMSCs transfected with miR-19b mimics.The morphological changes of BMSCs in the miR-19b mimics group induced by 5-aza were the largest,the cell volume was larger,and the cell aggregation was like vortex,while the cells in the miR-19b inhibitor group had no obvious morphological changes.Compared with control group,the expression levels of ANP,cTnI and cTnT in miR-19b mimics group significantly increased,while the proteins expressions in miR-19b inhibitor group significantly decreased.Conclusions MiR-19b has no ability to induce BMSCs differentiation,but it can improve the differentiation level of BMSCs to cardiomyocyte-like cells with 5-aza.
作者
周琦
明强
刘敏
骆春艳
ZHOU Qi;MING Qiang;LIU Min;LUO Chun-yan(Tianyou Hospital Affiliated to Wuhan University of Science and Technology,Wuhan 430064,China;Shanghai Tenth People′s Hospital,Shanghai 200072,China)
出处
《岭南心血管病杂志》
CAS
2021年第5期587-592,共6页
South China Journal of Cardiovascular Diseases
作者简介
周琦(1977-),女,硕士,副主任医师,研究方向为心力衰竭和高血压;通信作者:明强,E-mail:eep1002@aliyun.com。
