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目的:探讨乙醇对心肌祖细胞毒性作用的表观遗传机制。方法:以心肌祖细胞株为研究对象,用200mmol/L乙醇和溶解于该浓度乙醇的不同浓度姜黄素(5μmol/L,15μmol/L,25μmol/L)处理。Western blot检测组蛋白H3K9乙酰化水平,筛选出姜黄素的有效干预浓度,实时定量PCR检测心脏发育相关基因GATA4,Mef2c,Tbx5mRNA表达量的变化。结果:200mmol/L乙醇使组蛋白H3K9乙酰化水平升高2.76倍(P<0.05),同时使心脏发育相关基因GATA4,Mef2c,Tbx5表达均增加,其中GATA4,Mef2c较对照组有统计学差异(P<0.05)。5μmol/L、15μmol/L姜黄素分别使组蛋白H3K9乙酰化水平升高2.22、1.58倍(P<0.05);25μmol/L姜黄素使组蛋白H3K9乙酰化水平降至对照组水平,同时使心脏发育相关基因GATA4,Mef2c,Tbx5表达较乙醇组均下降,且与对照组无差异(P>0.05)。结论:姜黄素可干预乙醇对心肌祖细胞的高乙酰化失衡以及进而发生的相关基因的表达异常,可为了解乙醇致畸的机制以及临床预防先天性心脏病提供新的思路。
Objective: To investigate the epigenetic mechanism of ethanol toxicity on cardiac progenitor cells. Methods: Myocardial progenitor cell line was treated with 200 mmol / L ethanol and different concentrations of curcumin (5μmol / L, 15μmol / L, 25μmol / L) dissolved in this concentration of ethanol. The level of histone H3K9 acetylation was detected by Western blot. The effective intervention concentration of curcumin was screened. The expression of GATA4, Mef2c and Tbx5 mRNA in cardiac development was detected by real-time quantitative PCR. Results: The acetylation level of histone H3K9 increased by 2.76-fold (P <0.05) with 200mmol / L ethanol, while the expressions of GATA4, Mef2c and Tbx5 in cardiac development increased at the same time. The levels of GATA4 and Mef2c were significantly different from those in control group P <0.05). Curcumin at concentrations of 5μmol / L and 15μmol / L increased histone H3K9 acetylation levels by 2.22 and 1.58 folds (P <0.05), while curcumin at 25μmol / L reduced histone H3K9 acetylation levels to the control group The expression of GATA4, Mef2c and Tbx5 in development related genes decreased compared with those in ethanol group (P> 0.05). CONCLUSION: Curcumin can interfere with the abnormal acetylation of myocardial progenitor cells induced by ethanol and the abnormal expression of related genes, which may provide a new idea for understanding the mechanism of ethanol teratogenicity and preventing congenital heart disease clinically.