时孝晴,揭立士,吴 鹏,茆 军,张农山,王培民,殷松江.SS-31通过调控线粒体功能延缓细胞衰老的分子机制[J].,2022,(10):1834-1840 |
SS-31通过调控线粒体功能延缓细胞衰老的分子机制 |
The Molecular Mechanism of SS-31 Mediating Mitochondrial Function to Delay the Senescence of HEK293T Cells |
投稿时间:2021-10-26 修订日期:2021-11-21 |
DOI:10.13241/j.cnki.pmb.2022.10.008 |
中文关键词: SS-31 线粒体功能 细胞衰老 |
英文关键词: SS-31 Mitochondrial function Cell senescence |
基金项目:江苏省自然科学基金青年项目(SBK2020041404);国家自然科学基金青年项目(82074460);江苏省研究生科研与实践创新计划项目(KYCX21_1684);江苏省高校自然科学基金面上项目(20KJB360003) |
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中文摘要: |
摘要 目的:探讨SS-31通过调控线粒体功能延缓细胞衰老的调控效应。方法:将HEK293T细胞分为空白组、模型组、SS-31组。使用H2O2诱导应激性衰老模型,然后采用SS-31对HEK293T细胞进行干预。SA-β-gal衰老染色试剂盒检测细胞衰老水平;JC-1试剂盒检测细胞线粒体膜电位水平;荧光倒置显微镜观察线粒体活性氧(ROS)荧光强度;ATP检测试剂盒检测细胞内ATP水平;蛋白质免疫印迹检测P53,P21,Acetyl-p53,Sirt1蛋白表达。结果:模型组SA-β-gal阳性率高于空白组,应用SS-31后阳性率显著降低,差异具有统计学意义(P<0.05);ROS荧光检测:与空白组比较,模型组荧光强度上升,而SS-31组荧光强度较模型组下降,差异有统计学意义(P<0.05);线粒体膜电位检测:与空白组比较,模型组红色荧光强度显著下降,SS-31干预后,红色荧光强度显著上升,差异有统计学意义(P<0.05);ATP检测:模型组ATP水平低于对照组,SS-31组高于模型组,差异有统计学意义(P<0.05);此外,模型组P53,P21及Acetyl-p53蛋白表达水平较空白组增加(P<0.05),而SS-31组较模型组有所降低,差异有统计学意义(P<0.05),模型组Sirt1蛋白表达水平较空白组降低(P<0.05),而SS-31组较模型组升高,差异有统计学差异(P<0.05)。结论:SS-31能够通过改善线粒体功能延缓HEK293T细胞衰老。 |
英文摘要: |
ABSTRACT Objective: To investigate the regulatory effect of SS-31 in delaying cell senescence through mediating mitochondrial function. Methods: The HEK293T cell were divided into blank group, model group and SS-31 group. Use H2O2 to induce a stress-induced aging model, and then use SS-31 to intervene in HEK293T cell. SA-β-gal senescence staining kit detects cell senescence level; JC-1 kit detects cell mitochondrial membrane potential level; fluorescence inverted microscope observes mitochondrial reactive oxygen species (ROS) fluorescence intensity; ATP detection kit detects intracellular ATP level; protein Western blotting was used to detect the expression levels of P53, P21, Acetyl-p53 and Sirt1. Results: The percentage of SA-β-gal positive cells in the model group was higher than that of the blank group, and the percentage of SA-β-gal positive cells in the SS-31 group was reduced, and the difference was statistically significant (P<0.05); ROS fluorescence detection found that the model group produced a lot of Green fluorescence, while the fluorescence intensity of the SS-31 group decreased, and the difference was statistically significant (P<0.05); mitochondrial membrane potential detection showed that the red fluorescence intensity of the model group decreased significantly compared with the blank group. After the intervention of SS-31, the red fluorescence The intensity increased significantly, and the difference was statistically significant (P<0.05); ATP detection showed that the ATP level of the model group was lower than that of the control group, and the SS-31 group was higher than the model group, and the difference was statistically significant (P<0.05); The protein expression levels of P53, P21 and Acetyl-p53 increased compared with the blank group (P<0.05), while the SS-31 group was lower than the model group. The difference was statistically significant (P<0.05). The expression level of Sirt1 protein in the model group was higher than that of the blank group. The blank group decreased (P<0.05), while the SS-31 group was higher than the model group, the difference was statistically significant (P<0.05). Conclusion: SS-31 can improve the function of mitochondria, thereby delaying the senescence of HEK293T cell. |
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