Department of Pediatric Cardiac Surgery Center, Beijing Anzhen Hospital, Capital Medical University, Beijing 100029, China.
Chin Med J (Engl). 2017 Oct 5;130(19):2346-2353. doi: 10.4103/0366-6999.215328.
Current knowledge indicates that oxidative damage and the following inflammation is pivotal pathway for myocardial cell death. In recent decades, hydrogen sulfide (H2S) has been identified as a novel endogenous vasodilator and neuromodulator due to its antioxidation capacity. However, whether H2S pretreatment in neonatal mouse cardiomyocytes is a protection effect against oxidative stress remains elusive.
Primary neonatal mouse cardiomyocytes were isolated and cultured, subsequently, pretreated with the H2S donor, sodium hydrosulfide (NaHS). Cell viability, lactate dehydrogenase (LDH) release, and reactive oxygen species (ROS) production are evaluated. The levels of superoxide dismutase (Sod2) and Sirtuin 1 (Sirt1), a deacetylation enzyme, were detected by Western blotting. The statistics was performed using independent-sample t-test.
NaHS (100 μmol/L) had no toxicity to primary neonatal mouse cardiomyocytes. Furthermore, NaHS pretreatment significantly improved neonatal mouse cardiomyocytes survival after H2O2-induced cell death, indicated by the decrease in LDH release (40.00 ± 2.65% vs. 65.33 ± 4.33%, P < 0.01) and ROS production (1.90 ± 0.33 vs. 4.56 ± 0.56, P < 0.05), and that the salubrious effect was accompanied by the upregulation of Sod2 expression. In addition, the study showed that NaHS pretreatment improved mitochondrial DNA number in neonatal mouse cardiomyocyte. Furthermore, the result demonstrated NaHS increased the expression of Sirt1 in neonatal mouse cardiomyocyte. Ex 527, an inhibitor of Sirt1, could attenuate these effects of NaHS-induced Sod2 expression and mtDNA number increase, furthermore, abrogate the cytoprotective effects of NaHS for neonatal mouse cardiomyocytes.
Sirt1 mediated H2S-induced cytoprotection effects in neonatal mouse cardiomyocytes.
目前的知识表明，氧化损伤和随之而来的炎症是心肌细胞死亡的关键途径。近几十年来，由于具有抗氧化能力，硫化氢（H2S）已被确定为一种新型内源性血管舒张剂和神经调节剂。然而，H2S 预处理是否对氧化应激具有保护作用，在新生鼠心肌细胞中仍不得而知。
分离并培养原代新生鼠心肌细胞，然后用 H2S 供体硫氢化钠（NaHS）预处理。通过检测细胞活力、乳酸脱氢酶（LDH）释放和活性氧（ROS）产生来评估。通过 Western blot 检测超氧化物歧化酶（Sod2）和去乙酰化酶 Sirtuin 1（Sirt1）的水平。使用独立样本 t 检验进行统计学分析。
NaHS（100 μmol/L）对原代新生鼠心肌细胞没有毒性。此外，NaHS 预处理可显著改善 H2O2 诱导的细胞死亡后新生鼠心肌细胞的存活率，表现为 LDH 释放（40.00±2.65% vs. 65.33±4.33%，P<0.01）和 ROS 产生（1.90±0.33 vs. 4.56±0.56，P<0.05）减少，这种有益作用伴随着 Sod2 表达的上调。此外，研究表明，NaHS 预处理可增加新生鼠心肌细胞中线粒体 DNA 的数量。此外，结果表明，NaHS 可增加新生鼠心肌细胞中 Sirt1 的表达。Sirt1 抑制剂 Ex 527 可减弱 NaHS 诱导的 Sod2 表达和 mtDNA 数量增加的作用，并消除 NaHS 对新生鼠心肌细胞的保护作用。
Sirt1 介导 H2S 诱导的新生鼠心肌细胞保护作用。
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