Department of Cardiovascular Surgery, The First Affiliated Hospital of Soochow University, Wansheng Street, Suzhou Industrial District, Suzhou, China.
Department of Thoracic Surgery, The First Affiliated Hospital of Soochow University, Suzhou, China.
J Biol Regul Homeost Agents. 2021 Jan-Feb;35(1):85-96. doi: 10.23812/20-101-A.
This study aimed at probing into the function of muscone in ameliorating myocardial ischemiareperfusion (I/R) injury and exploring the underlying mechanism. To analyze the function of muscone, H9c2 cardiomyocytes were treated with hypoxia/reoxygenation (H/R) and Sprague-Dawley (SD) rats were treated with left anterior descending (LAD) of the coronary artery ligation for 30 min and reperfusion for 2 h to induce myocardial I/R injury. Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot were used to detect the expression of SIRT3. MTT assay and TUNEL assay were performed to investigate H9c2 viability and apoptosis, respectively. ELISA was employed to determine the expressions of inflammatory cytokines TNF-α, IL-6 and IL-1β, and myocardial injury markers CK and LDH. Oxidative stress markers MDA and SOD, and ROS expression levels were also detected. SIRT3 inhibitor 3-TYP was used to further confirm whether muscone worked via the augmentation of SIRT3. Herein, we found that muscone significantly inhibited inflammation and oxidative stress in H9c2 cardiomyocytes in a dose-dependent manner. H9c2 viability was promoted by muscone while apoptosis was inhibited. In SD rats, pre-treatment of muscone alleviated I/R injury-induced cardiac function dysregulation and left ventricle remolding. Furthermore, muscone increased SIRT3 expression at both mRNA and protein levels. With 3-TYP inhibiting SIRT3, the protective effects of muscone in H9c2 cardiomyocytes and SD rats were all significantly alleviated. In summary, muscone can attenuate inflammation, oxidative stress and cardiomyocytes injury in H9c2 cells treated with H/R and alleviate myocardial I/R injury of SD rats, which are dependent on SIRT3.
本研究旨在探讨麝香酮改善心肌缺血再灌注（I/R）损伤的作用，并探讨其潜在机制。为了分析麝香酮的功能，用缺氧/复氧（H/R）处理 H9c2 心肌细胞，用左前降支（LAD）冠状动脉结扎 30 min 并再灌注 2 h 处理 Sprague-Dawley（SD）大鼠，以诱导心肌 I/R 损伤。用定量实时聚合酶链反应（qRT-PCR）和 Western blot 检测 SIRT3 的表达。用 MTT assay 和 TUNEL assay 分别检测 H9c2 细胞活力和细胞凋亡。用 ELISA 测定炎症细胞因子 TNF-α、IL-6 和 IL-1β以及心肌损伤标志物 CK 和 LDH 的表达。还检测了氧化应激标志物 MDA 和 SOD 以及 ROS 表达水平。用 SIRT3 抑制剂 3-TYP 进一步证实麝香酮是否通过增强 SIRT3 发挥作用。在此，我们发现麝香酮以剂量依赖性方式显著抑制 H9c2 心肌细胞中的炎症和氧化应激。麝香酮促进 H9c2 细胞活力，抑制细胞凋亡。在 SD 大鼠中，麝香酮预处理减轻了 I/R 损伤引起的心脏功能失调和左心室重塑。此外，麝香酮增加了 SIRT3 在 mRNA 和蛋白水平的表达。用 3-TYP 抑制 SIRT3 后，麝香酮在 H9c2 心肌细胞和 SD 大鼠中的保护作用均明显减轻。总之，麝香酮可减轻 H/R 处理的 H9c2 细胞中的炎症、氧化应激和心肌细胞损伤，并减轻 SD 大鼠的心肌 I/R 损伤，这依赖于 SIRT3。
