Tian Kejun, Song Lijuan, Liu Liping, Lai Tao, Liu Wenxia
Department of Cardiology, The First Affiliated Hospital of Gannan Medical University, Ganzhou, Jiangxi Province 341000, PR China.
Department of Cardiology, Ruijin People's Hospital, Ruijin, Jiangxi Province 342500, PR China.
ACS Omega. 2025 May 16;10(21):21777-21785. doi: 10.1021/acsomega.5c01408. eCollection 2025 Jun 3.
This study aimed to determine the protective role and mechanism of rutin on myocardial ischemia-reperfusion (I/R) injury. First, a myocardial I/R injury model was established and subjected to ischemia and hypoxia to simulate the model of I/R injury in vivo. Triphenyltetrazolium chloride was used to determine the myocardial infarct area. Immunohistochemistry, the TUNEL assay, and hematoxylin and eosin staining were performed to determine NLRP3 expression, myocardial cell apoptosis, and cardiomyocyte morphology, respectively. Western blot analysis, the CCK-8 assay, and an ELISA were used to determine pyroptosis-associated protein expression, cell viability, and inflammatory factor expression, respectively. Rutin was shown to reduce the myocardial infarct area and myocardial cell apoptosis in vivo. Furthermore, rutin reduced the CK-MB, cTnT/I, and malondialdehyde levels after I/R injury. NLRP3 expression was downregulated after treatment with rutin compared to the sham group. NF-κB, NLRP3, cleaved caspase-1, and cleaved gasdermin D protein were also downregulated after rutin treatment. Rutin treatment enhanced cell viability after hypoxia-reperfusion conditions in experiments, reduced cell apoptosis, and reduced the LDH and ROS levels. Rutin also downregulated the expression of inflammatory factors (IL-1β, TNF-α, and IL-18) and pyroptosis-related proteins (NF-κB, NLRP3, cleaved caspase-1, and cleaved gasdermin D). Moreover, the protective effect of rutin on myocardial I/R injury disappeared after the NLRP3 agonist, nigericin, combined with rutin. These findings indicated that rutin protected myocardial I/R injury via the NF-κB/NLRP3/pyroptosis pathway and suggested a new treatment approach for myocardial I/R injury.
本研究旨在确定芦丁对心肌缺血再灌注（I/R）损伤的保护作用及其机制。首先，建立心肌I/R损伤模型，使其遭受缺血和缺氧以模拟体内I/R损伤模型。使用氯化三苯基四氮唑来确定心肌梗死面积。分别进行免疫组织化学、TUNEL检测以及苏木精-伊红染色，以确定NLRP3表达、心肌细胞凋亡和心肌细胞形态。分别使用蛋白质免疫印迹分析、CCK-8检测和酶联免疫吸附测定来确定焦亡相关蛋白表达、细胞活力和炎症因子表达。结果显示，芦丁可减小体内心肌梗死面积并减少心肌细胞凋亡。此外，芦丁可降低I/R损伤后的肌酸激酶同工酶（CK-MB）、心肌肌钙蛋白T/I（cTnT/I）和丙二醛水平。与假手术组相比，芦丁治疗后NLRP3表达下调。芦丁治疗后，核因子κB（NF-κB）、NLRP3、裂解的半胱天冬酶-1和裂解的gasdermin D蛋白也下调。在实验中，芦丁治疗可增强缺氧复灌注条件下的细胞活力，减少细胞凋亡，并降低乳酸脱氢酶（LDH）和活性氧（ROS）水平。芦丁还下调炎症因子（白细胞介素-1β、肿瘤坏死因子-α和白细胞介素-18）和焦亡相关蛋白（NF-κB、NLRP3、裂解的半胱天冬酶-1和裂解的gasdermin D）的表达。此外，NLRP3激动剂尼日利亚菌素与芦丁联合使用后，芦丁对心肌I/R损伤的保护作用消失。这些研究结果表明，芦丁通过NF-κB/NLRP3/焦亡途径保护心肌I/R损伤，并为心肌I/R损伤提出了一种新的治疗方法。
