鹿茸多肽诱导心肌干细胞向心肌细胞分化的机制研究
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篇名: 鹿茸多肽诱导心肌干细胞向心肌细胞分化的机制研究
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目的:探讨鹿茸多肽诱导心肌干细胞(CSCs)向心肌细胞分化的机制。方法:从SD大鼠体内分离CSCs,鉴定后培养传代至P3代后,分为空白对照组、5-氮胞苷诱导组(3 μmol/L)、鹿茸多肽诱导组(800 μg/ml)及其联合诱导组,除空白对照组细胞只加含3%胎牛血清的培养液外,其余各组细胞加入含相应诱导剂的培养液,培养14 d。采用免疫组化法检测细胞表面心肌肌钙蛋白T(cTnT)表达,免疫印迹技术检测细胞cTnT、活化复制因子2(ATF-2)和肌细胞增强因子2C(MEF-2C)表达,实时定量聚合酶链式反应技术检测细胞cTnT和转录因子Nkx2.5、GATA4 mRNA表达。结果:成功制得CSCs,纯度>97%。与空白对照组(cTnT呈阴性)比较,各诱导组细胞cTnT呈阳性,cTnT、ATF-2、MEF-2C蛋白表达和cTnT、Nkx2.5、GATA4 mRNA表达均增强(P<0.05)。与5-氮胞苷诱导组比较,鹿茸多肽诱导组细胞cTnT、MEF-2C蛋白表达和cTnT、GATA4 mRNA表达均增强(P<0.05),联合诱导组细胞cTnT、ATF-2、MEF-2C蛋白表达和cTnT、GATA4 mRNA表达均增强(P<0.05)。结论:鹿茸多肽可能通过增强Nkx2.5、GATA4、ATF-2、MEF-2C等转录因子的表达来诱导CSCs向心肌细胞分化。

ABSTRACT: OBJECTIVE: To explore the mechanism of differentiation from cardiac stem cells (CSCs) into cardiac cells induced by Cornu Corvi polypeptides. METHODS: CSCs were isolated from SD rats, and cultured to P3 generation after identification and then divided into blank control group, 5-azacitidine induction group (3 μmol/L), Cornu Corvi polypeptides induction group (800 μg/ml) and combination induction group. Except blank control group was cultured with culture solution containing 3% FBS, other groups were cultured with culture solution containing relevant derivant for 14 d. The expression of cTnT was determined by immunohistochemical method; the expression of cTnT, ATF-2 and MEF-2C were determined by immunoblot assay; mRNA expression of cTnT, Nkx2.5 and GATA4 were detected by RT-PCR. RESULTS: CSCs were prepared successfully with purity>97%. Compared with blank control group (cTnT being negative), cTnT of induction groups were positive, and the protein expression of cTnT, ATF-2 and MEF-2C and mRNA expression of cTnT, Nkx2.5 and GATA4 were all increased (P<0.05). Compared with 5-azacitidine induction group, the protein expression of cTnT and MEF-2C and mRNA expression of cTnT and GATA4 were decreased in Cornu Corvi polypeptide groups (P<0.05); the protein expression of cTnT, ATF-2 and MEF-2C and mRNA expression of cTnT and GATA4 were all increased in combination induction group (P<0.05). CONCLUSIONS: Cornu Corvi polypeptides can induce differentiation of CSCs into cardiac cells by promoting the expression of Nkx2.5, GATA4, ATF-2, MEF-2C and other transcription factors.
期刊: 2016年27卷第34期
作者: 王艳玲,黄晓巍,李哲,何璐,徐岩,曲晓波
AUTHORS: WANG Yanling,HUANG Xiaowei,LI Zhe,HE Lu,XU Yan,QU Xiaob
关键字: 鹿茸多肽;心肌干细胞;心肌细胞;分化
KEYWORDS: Cornu Corvi polypeptides; Cardiac stem cells; Cardiac cells; Differentiation
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