白黎芦醇对心肌细胞去乙酰化酶1表达时序的影响

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。

Effects of resveratrol on the sequential expression of SIRT1 in cardiomyocytes

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.