黄山药总皂苷对晚期糖基化终产物诱导心肌老化的保护作用及机制

Protective effect of total saponins of dioscorea pathaica on cardiac senescence induced by advanced glycation end-products and its mechanism

摘要:

目的 研究黄山药总皂苷(TSDP)对晚期糖基化终产物(AGEs)诱导的心肌老化产生的作用并初步探讨其机制。方法 用不同浓度TSDP干预原代心肌细胞,以CCK-8试剂盒(CCK-8)检测细胞活性。细胞预先TSDP干预 2h后加入AGEs,通过免疫印迹实验检测沉默信息调节因子(SIRT3)、超氧化物歧化酶-2 (SOD2)、及衰老相关蛋白p53和p16水平的表达；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)。结果 与对照组比较,AGEs干预组β半乳糖苷酶(SA-β-Gal)活性增加,p53和p16蛋白水平增加, SOD2和SIRT3蛋白表达下降,ROS产生增多,差异有统计学意义(P<0.05)。与AGEs组相比,TSDP预干预组SA-β-Gal活性降低,SIRT3和SOD2蛋白水平增加,p53和p16下降,ROS增加,差异有统计学意义(P<0.05)。结论 TSDP对AGEs诱导的心肌细胞老化起保护作用,其机制可能与升高SIRT3和调节氧化应激有关。

Abstract:

Objective To investigate the effect of total saponins of Dioscorea pathaica (TSDP) on cardiac senescence induced by advanced glycation end-products (AGEs) and its mechanism. Methods Neonatal rat cardiomyocytes were incubated with different concentrations of TSDP, and cell viability was measured by Cell Counting Kit-8 (CCK-8). AGEs were added 2h after the cells were pre-treated with TSDP. Western blotting was used to detect levels of silent mating type information regulation 2 homolog-3 (SIRT3), superoxide dismutase-2 (SOD2), and aging-related p53 and p15. 2,7-Dichlorodi-hydrofluorescein diacetate (DCFH-DA) was used to detect intracellular reactive oxygen species (ROS). Results Compared with the control group, the activity of SA-β-Gal and the level of p53 and p16 increased, the production of ROS increased, and the expression of SOD2 and SIRT3 decreased in the AGEs group, the differences being statistically significant (P<0.05). Compared with the AGEs group, the activity of SA-β-Gal decreased, ROS increased, SIRT3 and SOD2 protein levels increased, and p53 and p16 decreased in the TSDP pre-treatment group, differences being statistically significant (P<0.05). Conclusion TSDP has protective effects on AGEs-induced cardiomyocyte aging, and its mechanism may be related to the increase of SIRT3 and regulation of oxidative stress.