Chen Xingcan, Sun Tong, Qi Yuxiang, Zhu Bingqi, Li Lan, Yu Jie, Ding Zhishan, Zhou Fangmei
School of Medical Technology and Information Engineering, Zhejiang Chinese Medical University, Hangzhou, Zhejiang 310053, China.
Puer Kunhong Biotechnology Company, Group C of Chamagu Town A, Simao District, Puer, Yunnan 665000, China.
Mol Immunol. 2025 Jan;177:32-43. doi: 10.1016/j.molimm.2024.12.003. Epub 2024 Dec 17.
Myocardial ischemia-reperfusion injury (MIRI) injury is a serious health problem, which can seriously affect the recovery of patients with myocardial infarction and even lead to death. Paeoniflorin (PF) is a potential therapeutic drug to prevent reperfusion injury. However, the mechanism of PF in MIRI is not clear. Compared with other cells, cardiomyocytes have the largest number of mitochondria. Therefore, this study researched the protective mechanism of paeoniflorin pretreatment on myocardial ischemia-reperfusion injury (AMI) from the perspective of mitochondrial autophagy. Paeoniflorin was given or not given to H9C2 cells 12 h before reperfusion. Pretreatment of paeoniflorin can significantly increase the viability of H9C2 cells and inhibit the increase of ROS secretion induced by OGD/R. The increase of MDC autophagy fluorescence and mitochondrial membrane potential (MMP) suggested that the myocardial protective effect of paeoniflorin may also be related to mitochondrial autophagy. Next, we detected the related signals in the classical mitochondrial autophagy pathway of PINK1/parkin by Q-PCR and Western blots. The results showed that the pretreatment of paeoniflorin could promote the levels of SIRT1, Beclin1, PINK1, parkin and LC3, inhibit the level of P62. In order to further clarify whether paeoniflorin-induced SIRT1 activation is necessary for autophagy and its potential mechanism, we detected the autophagy level of H9C2 cells with SIRT1 inhibitor (EX527). The results showed that after pretreatment of EX527, the protective effect of paeoniflorin on oxidative damage and autophagy pathway was significantly decreased. The mechanism may relate to SIRT1-PINK1/parkin mitochondrial autophagy pathway. In summary, these results suggested that paeoniflorin may protect H9C2 cells from OGD/R damage by activating SIRT1-PINK1/parkin pathway. This provides new experimental basis for paeoniflorin in the treatment of MIRI.
心肌缺血再灌注损伤（MIRI）是一个严重的健康问题，它会严重影响心肌梗死患者的康复，甚至导致死亡。芍药苷（PF）是一种预防再灌注损伤的潜在治疗药物。然而，PF在MIRI中的作用机制尚不清楚。与其他细胞相比，心肌细胞拥有数量最多的线粒体。因此，本研究从线粒体自噬的角度探讨了芍药苷预处理对心肌缺血再灌注损伤（AMI）的保护机制。在再灌注前12小时给予或不给予H9C2细胞芍药苷。芍药苷预处理可显著提高H9C2细胞的活力，并抑制OGD/R诱导的ROS分泌增加。MDC自噬荧光和线粒体膜电位（MMP）的增加表明，芍药苷的心肌保护作用可能也与线粒体自噬有关。接下来，我们通过Q-PCR和蛋白质免疫印迹法检测了经典线粒体自噬途径PINK1/parkin中的相关信号。结果表明，芍药苷预处理可促进SIRT1、Beclin1、PINK1、parkin和LC3的水平，抑制P62的水平。为了进一步阐明芍药苷诱导的SIRT1激活对于自噬是否必要及其潜在机制，我们用SIRT1抑制剂（EX527）检测了H9C2细胞的自噬水平。结果表明，EX527预处理后，芍药苷对氧化损伤和自噬途径的保护作用显著降低。其机制可能与SIRT1-PINK1/parkin线粒体自噬途径有关。综上所述，这些结果表明芍药苷可能通过激活SIRT1-PINK1/parkin途径保护H9C2细胞免受OGD/R损伤。这为芍药苷治疗MIRI提供了新的实验依据。
