摘要
背景:组蛋白乙酰化是诱导骨髓间充质干细胞向心肌样细胞分化的机制之一。目的:观察组蛋白去乙酰化酶抑制剂丁酸钠对体外培养的骨髓间充质干细胞向心肌样细胞分化的影响,并分析其诱导分化的机制。方法:分离、培养大鼠骨髓间充质干细胞,并进行鉴定。取第2代骨髓间充质干细胞,应用1mmol/L丁酸钠诱导其向心肌样细胞分化。结果与结论:实验分离培养的骨髓间充质干细胞高表达CD90,CD29;低表达CD45,CD31。加入丁酸钠后细胞增殖能力及组蛋白去乙酰化酶活性明显降低,但未发生明显凋亡现象,表明丁酸钠具有低毒性的特征。real-timePCR检测显示,经丁酸钠诱导后细胞GATA-4,MEF-2c,β-MHC等心肌细胞早期标志基因表达率明显增高(P<0.05)。Westernblot检测发现,经丁酸钠诱导后细胞心肌细胞特异性蛋白连接蛋白43和肌钙蛋白T的表达明显增高。免疫荧光测得的肌钙蛋白T表达结果与Westernblot一致。说明丁酸钠能够有效诱导骨髓间充质干细胞向心肌样细胞分化,其机制可能与抑制组蛋白去乙酰化酶活性,增强组蛋白的乙酰化有关。
BACKGROUND： Histone acetylation is one of mechanisms underlying induced cardiomyocyte differentiation of bone marrow mesenchymal stem cells （BMSCs）. OBJECTIVE： To investigate the effects of sodium butyrate, a histone deacetylate inhibitor, on differentiation of rat BMSCs into cardiomyocytes in vitro. METHODS： Rat bone marrow mesenchymal stem cells （rBM-MSCs） were isolated, cultured and identified. Passage 2 cells were induced to differentiate into cardiomyocytes by 1 mmol/L sodium butyrate. RESULTS AND CONLUSION： The isolated BMSCs were positive for CD90, but they were negative for CD31, CD45. After addition of sodium butyrate, cell proliferation ability and histone deacetylase activity were significantly decreased, but no obvious apoptosis was observed. Real-time PCR results showed that after addition of sodium butyrate, the expression level of myocardial genes including GATA-4, MEF-2c, β-MHC was significantly increased （P 〈 0.05）. Western blot results showed that after induced by sodium butyrate, the expression level of cardiac-specific proteins including connexin 43 and cardiac troponin T was significantly increased. Troponin T expression detected by immunofluorescence was consistent with that detected by western blot. These findings suggest that sodium butyrate can effectively induce BMSCs differentiation into cardiomyocytes in vitro. The mechanism may be related to inhibition of histone deacetylase activity and increase in histone acetylation.
出处
《中国组织工程研究》
CAS
CSCD
2012年第19期3462-3466,共5页
Chinese Journal of Tissue Engineering Research
