ATM; To investigate the effect of resveratrol on aged cardiomyocytes induced by D -galactose and the underlying mechanism. METHODS; Different concentrations of D - galactose (0, 0. 1, 1 , 10 and 100 g/L) were added into the culture medium for different processing time (0, 12, 24, 48 and 72 h) to induce cardiomyocyte senescence . Cell senescence was indentified by senescence - associated p - galactosidase staining. Cell viability was evaluated by MTT assay. FDG method was used to detect the quantity of β - galactosidase. The establishment of the cardiomyocyte aging mod -el was determined according to the outcome of these detections . After that, different concentrations of resveratrol (2, 10, 50 and 250 μnol/L) were added into the culture medium , and the effect of resveratrol on the aged cardiomyocytes was ob -served according to the outcome of FDG detection . The activity of superoxide dismutase (SOD) , the content of malondial-dehyde ( MDA) and the protein level of microtubule - related protein 1 light chain 3 ( LC3 ) in the cells were measured. RESULTS ; Compared with control group , the number of β -galactosidase - positive cells in aging group (10 g/L D - ga-lactose exposure for 48 h) was apparently increased. In aging group, the quantity of β - galactosidase detected by FDG was markedly increased , and the activity of SOD was down - regulated while the content of MDA in the cells was increased . The protein level of LC3 II/LC3 I was down -regulated. No significant change of the cell viability between the 2 groups was observed. Compared with aging group , the quantity ofβ - galactosidase in resveratrol treatment group ( at the concentrations of 10 |μmol/L and 50 μmol/L) was decreased. Resveratrol also up - regulated the activity of SOD and decreased the con -tent of MDA in the cells. The protein level of LC3 II/LC3 I was up - regulated by resveratrol. CONCLUSION: Activation of oxidative stress and down - regulation of autophagy exist in D - galactose - induced aged cardiomyocytes . Resveratrol attenuates D - galactose - induced senescence in cardiomyocytes .%目的:探讨白藜芦醇对D-半乳糖诱导的心肌细胞衰老的影响及其可能机制.方法:用不同浓度D-半乳糖(0、0.1、1、10和100g/L)作用不同时间(0、12、24、48和72 h)诱导原代心肌细胞衰老,通过β-半乳糖苷酶染色法鉴定细胞衰老,MTT法检测细胞活力,FDG法半定量检测β-半乳糖苷酶含量,构建体外心肌细胞衰老模型.根据FDG的结果来评估不同浓度白藜芦醇(2、10、50和250μmol/L)处理对衰老心肌细胞的影响,通过检测超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量及微管相关蛋白1轻链3(LC3)的变化来探讨白藜芦醇处理效应产生的可能机制.结果:与对照组相比,衰老组(10g/L D-半乳糖处理48 h)β-半乳糖苷酶染色阳性率明显增加,β-半乳糖苷酶含量明显增加;SOD活性下降,MDA含量增加;LC3Ⅱ/LC3Ⅰ水平下调;细胞活力无明显变化.与衰老组相比,白藜芦醇处理组(10μmol/L及 50μmol/L)β-半乳糖苷酶含量明显降低;SOD活性升高,MDA含量明显降低;LC3Ⅱ/LC3Ⅰ水平上调.结论:D-半乳糖诱导的心肌细胞衰老伴随着氧化应激的增强及自噬水平的下调;白藜芦醇能减轻D-半乳糖诱导的心肌细胞衰老的程度,其保护效应产生的机制可能与白藜芦醇抗氧化应激效应及上调自噬水平相关.
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