睾酮治疗延缓小鼠心肌细胞衰老:SIRT1保护作用  

Testosterone therapy delays murine cardiomyocyte aging: protective role of SIRT1

作　　者：雷达[1] 张莉[1] 朱桂平[1] 李博维[1] 

机构地区：[1]广东药学院附属第一医院心内科,广州510080

出　　处：《岭南心血管病杂志》2015年第3期407-411,共5页South China Journal of Cardiovascular Diseases

基　　金：广东省医学科学技术研究基金(项目编号:WSTJJ20131230440102196209093234)

摘　　要：目的观察睾酮是否通过SIRT1对衰老HL-1心肌细胞起保护作用。方法实验用HL-1心肌细胞系分为4组:对照组、百草枯(PQ)诱导衰老组、睾酮干预组、睾酮联合SIRT1抑制剂(EX-527)干预组。应用细胞化学染色法测定各组细胞衰老相关β-半乳糖苷酶(senescence-associatedβ-galactosidase,SAβ-gal)、2,7-二氯氢化荧光素二脂探针检测各组心肌细胞内活性氧(reactive oxygen species,ROS)水平、Western印迹检测各组心肌细胞SIRT1蛋白表达。结果与对照组相比,百草枯诱导可导致HL-1心肌细胞SAβ-gal染色体阳性比例增加(80.38%±8.94%vs.22.66%±5.11%,P<0.001)、ROS水平增加(荧光强度:4 875.71±240.58 vs.976.07±126.70,P<0.001)及SIRT1蛋白表达水平下调(0.31±0.03 vs.0.56±0.03,P<0.001),差异均有统计学意义。1μmol/L睾酮干预HL-1心肌细胞可逆转百草枯诱导的这一变化。与睾酮干预组相比,联合EX-527干预组HL-1心肌细胞SAβ-gal染色体阳性比例增加(71.64%±7.54%vs.34.82%±5.20%,P<0.001)、ROS水平升高(荧光强度:3 906.00±259.36 vs.1 465.76±323.61,P<0.001),差异均有统计学意义。结论 1μmol/L睾酮干预可延缓百草枯诱导的HL-1心肌细胞衰老,其保护作用是通过上调SIRT表达来实现的。Objectives To observe whether the feature in senescent HL-1 cardiomyocytes could be reversed by testosterone via up-regulation of SIRT1. Methods Cultured HL-1 cardiomyocytes were divided into 4 groups ： control group, senescent cells induced by paraquat（PQ） group, senescent cells treated by testosterone group and testosterone in combination with SIRT1 inhibitor（EX-527） group. Senescence-associated β-galactosidase（SAβ-gal） was measured by chemical staining, intracellular reactive oxygen species（ROS） level was detected by fluorescent probe, and expression of SIRT1 was detected by Western blot. Results Compared with control group, proportions of SAβ-gal positive cells and ROS levels significantly increased in HL-1 cardiomyocytes treated by PQ, while expressions of SIRT1 protein was down-regulated（SAβ-gal： 80.38% ±8.94% vs. 22.66% ±5.11%, P〈0.001; ROS ： 4 875.71 ±240.58 vs.976.07±126.70, P〈0.001; SIRT1 ： 0.31 ±0.03 vs. 0.56 ±0.03, P〈0.001）. These changes were improved in senescent cardiomyocytes treated by testosterone（1 μmol / L）. Proportions of SAβ-gal positive cells and ROS levels were higher in cardiomyocytes treated by testosterone in combination with EX-527 than by testosterone only（SAβ-gal：71.64%±7.54%vs. 34.82% ±5.20%, P〈0.001; ROS ： 3 906.00 ±259.36 vs. 1 465.76 ±323.61, P〈0.001）. Conclusions 1 μmol / L testosterone can delay HL-1 cardiomyocyte aging induced by PQ depending on the protective role of SIRT1.

关 键 词：衰老 睾酮 心肌细胞 SIRT1 

分 类 号：R33[医药卫生—人体生理学]