Sirt3在晚期糖基化终产物诱导的心肌老化中的作用及机制  

Role and mechanism of Sirt3 in advanced glycosylation end-products-induced cardiomyocyte aging

作　　者：徐婉莹 查志敏 汤金梅 Xu Wanying;Zha Zhimin;Tang Jinmei(Department of Gerontology,Suzhou Ninth People′s Hospital,Suzhou 215000,Jiangsu Province,China;Department of Gerontology,First Affiliated Hospital of Nanjing Medical University,Nanjing 210029,China)

机构地区：[1]苏州市第九人民医院老年医学科,江苏苏州215000 [2]南京医科大学第一附属医院老年医学科,南京210029

出　　处：《中华老年多器官疾病杂志》2023年第12期934-938,共5页Chinese Journal of Multiple Organ Diseases in the Elderly

基　　金：江苏省老年健康科研项目(LD2021030);吴江区科教兴卫项目(wwk201809)。

摘　　要：目的研究沉默信息调节因子3(Sirt 3)在晚期糖基化终产物(AGEs)诱导心肌老化过程中的作用,并初步探讨黄山药总皂苷(TSDP)改善心肌老化的机制。方法提取原代心肌细胞,AGEs干预诱导细胞老化,转染小干扰RNA(siRNA)敲降Sirt3表达。免疫印迹实验检测P16、P53、超氧化物歧化酶-2(SOD2)及SIRT3的蛋白表达水平。半乳糖苷酶(SA-β-Gal)染色试剂盒检测细胞老化。2′,7′-二氯二氢荧光素二乙酸酯探针(DCFH-DA)检测细胞内活性氧(ROS)。TSDP预干预AGEs诱导的心肌老化后,采用SA-β-Gal染色试剂盒检测细胞老化。采用GraphPad Prism统计软件进行数据分析。组间比较采用单因素方差分析。结果和未转染慢病毒的对照组相比,转染阴性参照NC siRNA后加入AGEs的干预组SIRT3及SOD2的表达减少,P53的表达增加,SA-β-Gal染色阳性细胞比例及活性氧的水平增加。转染Sirt 3 siRNA后加入AGEs的干预组与对照组相比,SIRT3及SOD2表达减少,SA-β-Gal染色阳性细胞比例及活性氧水平升高,且与转染Sirt 3 siRNA组相比,P53表达水平进一步增加。转染NC siRNA的TSDP预处理AGEs干预组较转染NC siRNA后AGEs干预组SA-β-Gal染色阳性细胞比例减少,但转染Sirt 3 siRNA的TSDP预处理AGEs干预细胞后上述TSDP引起的SA-β-Gal改变作用消失。结论AGEs可能是通过下调Sirt3的表达水平,减少抗氧化物蛋白SOD2的表达,加重线粒体氧化应激,促进AGEs诱导的心肌老化进程。TSDP可能通过调控Sirt3的表达水平而影响老化。Objective To study the role of silent mating type information regulation 2 homolog-3(Sirt 3)in the process of cardiac senescence induced by advanced glycosylation end-products(AGEs)and to explore the mechanism of total saponins extracted from Dioscorea panthaica(TSDP)in cardiac senescence.Methods After neonatal rat cardiomyocytes were isolated and cultured and identified,the cells were incubated with AGEs.Small interfering RNA(siRNA)was used to knock down the expression of Sirt3 in the cells.Western blotting was used to detect the expression of P16 and P53 and superoxide dismutase-2(SOD2).SA-β-Gal kit was employed to detect cardiomyocyte senescence,and 2,7-dichlorodi-hydrofluorescein diacetate(DCFH-DA)probe was used to detect intracellular reactive oxygen species(ROS).In the cells with TSDP pre-intervention after AGEs treatment,cardiac senescence was detected by SA-β-Gal kit.GraphPad Prism statistical analysis was performed.One-way ANOVA was conducted for data comparison between two groups.Results Compared with the control cells,the levels of SIRT3 and SOD2 were reduced,and that of P53 was increased,and the proportion of SA-β-Gal positive cells and production of ROS were elevated in the AGEs+negative control(NC)siRNA group.Same changing trends were observed in the AGEs+Sirt3 siRNA group,and the expression level of senescence-related protein P53 was also up-regulated.TSDP pre-intervention decreased the proportion of SA-β-Gal positive cells when compared with the AGEs+NC siRNA group,but the change was decreased after transfection with Sirt3 siRNA.Conclusion AGEs may decrease the expression of SOD2 by down-regulating Sirt3,then aggravate mitochondrial oxidative stress and promote cardiomyocyte aging.TSDP may affect cardiac senescence by regulating the expression of Sirt3.

关 键 词：晚期糖基化终产物 Sirt3 超氧化物歧化酶-2 活性氧 心肌 老化 

分 类 号：R541[医药卫生—心血管疾病]