白黎芦醇对心肌细胞去乙酰化酶1表达时序的影响

Effects of resveratrol on the sequential expression of SIRT1 in cardiomyocytes

目的探讨白黎芦醇(RES)在心肌细胞中对去乙酰化酶(SIRT)1表达时序的影响。方法大鼠心肌细胞株H9c2培养48h后,分别以0(对照组)、10(低剂量组)、20(中等剂量组)、50、100μmol/L(高剂量组)的RES处理0、12、24、36、48h,采用噻唑蓝(MTT)比色法测定细胞活力;反转录-聚合酶链反应(RT-PCR)分析SIRT1 mRNA表达情况,Western blot法检测SIRT1蛋白表达水平的变化。结果H9c2心肌细胞经RES处理后,MTT测得的吸光度(A)值在低、中剂量升高,20μmol/L组作用最显著(P<0.05),50μmol/L组开始下降,低于对照组(P>0.05),100μmol/L组降至最低(P<0.05);各组SIRT1 mRNA和蛋白表达A值均高于对照组(P<0.05),与低、高剂量组相比,中等剂量组表达最高(P<0.05);中等剂量组RES处理0、12、24、36、48h后,不同时间点SIRT1 mRNA和蛋白表达A值均高于对照组(P<0.05),以24h时表达最强(P<0.01)。结论20μmol/LRES作用24h时心肌细胞的SIRT1表达最强。

Objective To observe the effect of resveratrol （RES）, an activator of silent information regulator 2, on the sequential expression of SIRTI in cardiomyocytes. Methods After 48 h culture under normal condition, the embryonal rat heart-derived H9c2 cells were treated with 0 （control group）, 10 （low dose group）, 20 （medium dose group） 50 and 100 μmol/L （high dose groups） of RES for 0, 12, 24, 36 and 48 h. Cell viability was analyzed by MTr assay, expression of SIRT1 mRNA by RT-RCR and SIRT1 products by Western blot. Results The optical densities （A） of MTT was increased in the low and medium dose groups, appeared markedly higher with 20 μmol/L RES ment （P〈0.05）, and was lower than normal in the high dose groups, particularly in the 100 μmol/L group （P〈O.05）. The expressions of SIRT1 mRNA and protein were increased after treated with different concentrations of RES （P〈0.05）, most notability with 20μmol/L. At different time spots of incubation with 20 p.mol/L RES （0, 12, 24, 36, and 48 h）, the A values of SIRT1 mRNA and proteins were higher than those in controls （P〈0.05）, with a peak at 24 h. Conclusion The maximum expression of SIRTI was achieved with treatment of 20μmol/L RES for 24 h in cadiacmyocytes.