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作 者:曾缨[1] 郑民缨[1] 丰岩清[1] 冯慧宇[1] 张成[1]
机构地区:[1]中山大学附属第一医院神经科,广州510080
出 处:《中国神经精神疾病杂志》2009年第7期398-401,共4页Chinese Journal of Nervous and Mental Diseases
基 金:国家自然科学基金(编号:30370510);广东省自然科学基金博士启动项目(编号:07301430);卫生部临床重点项目(编号:2001321)
摘 要:目的探讨组蛋白去乙酰化酶(histonedeacetylase,HDAC)抑制剂丁酸钠和维甲酸体外诱导大鼠骨髓间质干细胞(rat mesenchymal stem cells,rMSC)分化为肌样细胞的作用。方法分离、培养、增殖rMSC,用HDAC抑制剂丁酸钠和维甲酸诱导rMSC分化为肌样细胞,免疫细胞化学方法检测诱导前后细胞肌肉特异性蛋白desmin和α-sarcomeric actin的表达,RT-PCR方法检测诱导前后骨骼肌特异性转录因子MyoD、myogenin、MRF 4和肌肉特异性肌酸磷酸激酶(muscle specific creatine kinase,MCK)mRNA的表达。结果rMSC诱导后可见多核肌管样细胞,且肌肉特异性蛋白desmin和α-sarcomeric actin的表达呈阳性。MyoD基因mRNA在诱导前后均有表达,但诱导1 d后表达上调,差异有统计学意义(P<0.05);myogenin和MRF 4基因mRNA在诱导前无表达,诱导1 d后开始表达,MCK基因mRNA在诱导7 d后开始表达。结论HDAC抑制剂丁酸钠和维甲酸可诱导rMSC分化成肌样细胞,并上调骨骼肌特异性转录因子和肌肉特异性肌酸磷酸激酶的表达。Objective To induce rat mesenchymal stem cells (rMSC) to differentiate into muscle-like cells with HDAC inhibitor sodium butyrate and retinoid acid. Methods Rat MSCs were isolated, cultured, proliferated and induced to differentiate into muscle-like cells with HDAC inhibitor sodium butyrate and retinoid acid. The expressions of muscle specific protein desmin and α-sarcomeric actin in muscle-like cells and undifferentiated rMSC were detected by immunocytochemistry. The mRNA expressions of skeletal muscle specific transcription factors MyoD, myogenin, MRF4 and MCK were detected by reverse transcription polymerase chain reaction (RT-PCR). Results Muhinucleated myotube-like cells ap- peared after induction. The muscle markers of desmin and α-sarcomeric actin were positive in muscle-like ceils. Both rMSC and muscle-like cells expressed MyoD mRNA. But the expression level of MyoD mRNA was significantly increased only in the muscle-like cells ( P 〈 0. 05 ). RMSCs began to express myogenin and MRF4 mRNA 1 d after induction and began to express MCK mRNA 7 d after induction. Conclusions Rat MSCs are able to differentiate into muscle-like cells under treatment with HDAC inhibitor sodium butyrate and retinoid acid. The mRNA expressions of skeletal muscle specific transcription factors and MCK are upregulated in muscle-like ceils.
关 键 词:干细胞 肌肉 组蛋白去乙酰化酶抑制剂 维甲酸 肌营养不良症
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