Sulforaphane prevents rat cardiomyocytes from hypoxia/reoxygenation injury in vitro via activating SIRT1 and subsequently inhibiting ER stress
Sulforaphane prevents rat cardiomyocytes from hypoxia/reoxygenation injury in vitro via activating SIRT1 and subsequently inhibiting ER stress
复制标题
萝卜硫素通过激活 SIRT1 并随后抑制 ER 应激，在体外预防大鼠心肌细胞缺氧/复氧损伤
DOI：
10.1038/aps.2015.130
复制
发表时间：
2016-03-01
影响因子：
8.2
通讯作者：
Chen, Ai-hua
中科院分区：
文献类型：
--
作者：
Li, Yun-peng;Wang, Shu-lin;Chen, Ai-hua
目的：萝卜硫醚(SFN)是一种天然的饮食异硫氰酸酯，对心血管疾病具有良好的治疗作用。本研究旨在探讨三七总皂苷对体外培养的大鼠心肌缺氧/复氧(H/R)损伤的保护作用机制。方法：体外培养的乳鼠心肌细胞经三七总皂苷预处理后，先缺氧3h，再复氧3h。检测细胞存活率和细胞凋亡率。测定caspase-3活性和线粒体膜电位(Delta Psim)。Western印迹分析内质网应激相关细胞凋亡蛋白的表达。结果：SFN(0.1~5mU·mol/L)可剂量依赖性地提高心肌细胞存活率，减少细胞凋亡率，抑制caspase-3活性。同时，三七总皂苷可明显减轻心肌细胞Delta psi的损伤，降低内质网应激相关的凋亡蛋白(GRP78、CHOP和caspase-12)的表达，上调心肌细胞SIRT1的表达和Bcl2/Bax比值。SIRT1特异性抑制剂Ex-527(1mU/L)可阻断SFN对心肌细胞的保护作用。结论：SFN可能通过激活SIRT1通路，抑制内质网应激依赖性细胞凋亡，从而减轻心肌细胞的缺氧再灌注损伤。
Aim: Sulforaphane (SFN), a natural dietary isothiocyanate, is found to exert beneficial effects for cardiovascular diseases. This study aimed to investigate the mechanisms underlying the protective effects of SFN in a model of myocardial hypoxia/reoxygenation (H/R) injury in vitro.Methods: Cultured neonatal rat cardiomyocytes pretreated with SFN were subjected to 3-h hypoxia followed by 3-h reoxygenation. Cell viability and apoptosis were detected. Caspase-3 activity and mitochondrial membrane potential (Delta psi m) was measured. The expression of ER stress-related apoptotic proteins were analyzed with Western blot analyses. Silent information regulator 1 (SIRT1) activity was determined with SIRT1 deacetylase fluorometric assay kit.Results: SFN (0.1-5 mu mol/L) dose-dependently improved the viability of cardiomyocytes, diminished apoptotic cells and suppressed caspase-3 activity. Meanwhile, SFN significantly alleviated the damage of Delta psi m and decreased the expression of ER stress-related apoptosis proteins (GRP78, CHOP and caspase-12), elevating the expression of SIRT1 and Bcl-2/Bax ratio in the cardiomyocytes. Co-treatment of the cardiomyocytes with the SIRT1-specific inhibitor Ex-527 (1 mu mol/L) blocked the SFN-induced cardioprotective effects.Conclusion: SFN prevents cardiomyocytes from H/R injury in vitro most likely via activating SIRT1 pathway and subsequently inhibiting the ER stress-dependent apoptosis.