Li Yongjun, Chen Xi, Yang Mingming, Yan Changshun, Zhang Rui, Zhang Xiaoguo, Zuo Pengfei, Carvalho Abdlay, Cao Guiqiu, Ma Genshan
Department of Cardiology, Zhongda Hospital, School of Medicine, Southeast University 87 Dingjiaqiao, Nanjing 210009, PR China; Department of Cardiology, The Fifth Affiliated Hospital of Xinjiang Medical University, Urumqi 830000, PR China.
Anqing First People's Hospital of Anhui Medical University, Anqing 246000, PR China.
Cell Signal. 2025 Nov;135:112045. doi: 10.1016/j.cellsig.2025.112045. Epub 2025 Aug 4.
The protective mechanism of tetramethylpyrazine (TMP), an alkaloid derived from the Chinese herbal remedy Ligusticum chuanxiong, in relation to myocardial ischemia/reperfusion injury (MIRI) is not yet fully understood. To investigate the therapeutic effects of TMP, we established an in vivo rat model of MIRI and an in vitro hypoxia and reoxygenation (H/R) model using H9c2 cells. First, TMP significantly reduced the infiltration of inflammatory cells in the myocardium, decreased serum levels of myocardial injury markers (CK-MB, cTnT and LDH), and alleviated myocardial infarct size in I/R rats. Additionally, TMP attenuated cell apoptosis and enhanced cell viability in H9c2 cells subjected to H/R. Furthermore, we employed NOD-like receptor protein 3 (NLRP3)-specific inhibitors (MCC950) and agonists (nigericin) to explore the role of the NLRP3 inflammasome in TMP's protective effects against MIRI. TMP was found to inhibit NLRP3 inflammasome activation and reduce caspase-1-dependent pyroptosis. We investigated the potential mechanism underlying the protective effect of TMP on MIRI using drug affinity responsive target stability (DARTS) and liquid chromatography tandem mass spectrometry (LC-MS/MS) techniques. It was found that myosin light chain-2 (Myl2) was the molecule directly responsible for TMP's protective effect on MIRI. Following TMP treatment, the protein levels of Myl2 increased in the heart tissues of the I/R rats and in H9c2 cells subjected to H/R in a dose-dependent manner. Finally, the cardioprotective effects of TMP, including the inhibition of the NLRP3 inflammasome, were partially negated by the genetic silencing of Myl2 using siRNA. This study indicated that TMP mitigated MIRI by regulating the inhibition of the Myl2-mediated NLRP3 signaling pathway.
川芎嗪（TMP）是一种源自中药川芎的生物碱，其对心肌缺血/再灌注损伤（MIRI）的保护机制尚未完全明确。为研究TMP的治疗效果，我们建立了大鼠MIRI体内模型以及使用H9c2细胞的体外缺氧复氧（H/R）模型。首先，TMP显著减少了心肌中炎性细胞的浸润，降低了血清中心肌损伤标志物（CK-MB、cTnT和LDH）的水平，并减轻了I/R大鼠的心肌梗死面积。此外，TMP减轻了H/R处理的H9c2细胞的凋亡并增强了细胞活力。进一步地，我们使用NOD样受体蛋白3（NLRP3）特异性抑制剂（MCC950）和激动剂（尼日利亚菌素）来探究NLRP3炎性小体在TMP对MIRI保护作用中的作用。发现TMP可抑制NLRP3炎性小体的激活并减少半胱天冬酶-1依赖性细胞焦亡。我们使用药物亲和响应靶点稳定性（DARTS）和液相色谱串联质谱（LC-MS/MS）技术研究了TMP对MIRI保护作用的潜在机制。发现肌球蛋白轻链-2（Myl2）是TMP对MIRI保护作用的直接负责分子。TMP处理后，I/R大鼠心脏组织和H/R处理的H9c2细胞中Myl2的蛋白水平呈剂量依赖性增加。最后，使用siRNA对Myl2进行基因沉默部分抵消了TMP的心脏保护作用，包括对NLRP3炎性小体的抑制。本研究表明，TMP通过调节对Myl2介导的NLRP3信号通路的抑制来减轻MIRI。
