Yin Suyue, Han Kaixi, Wu Di, Wang Zihan, Zheng Ruifang, Fang Lianhua, Wang Shoubao, Xing Jianguo, Du Guanhua
Beijing Key Laboratory of Drug Targets Identification and Drug Screening, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
Xinjiang Key Laboratory of Uygur Medical Research, Xinjiang Institute of Materia Medica, Urumqi, China.
Front Pharmacol. 2024 Oct 23;15:1423053. doi: 10.3389/fphar.2024.1423053. eCollection 2024.
Tilianin, a flavonoid compound derived from L., is recognized for its diverse biological functionalities, in particular alleviating myocardial ischemia-reperfusion injury (MIRI). There is ample evidence suggesting that the NLRP3 inflammasome has a significant impact on the development of MIRI. In this study, rats undergoing the ligation and subsequent release of the left anterior descending (LAD) coronary artery and H9c2 cardiomyocytes subjected to oxygen-glucose deprivation/reoxygenation (OGD/R) were used to investigate the effects of tilianin on NLRP3 inflammasome and its anti-MIRI mechanisms. Upon reperfusion, the rats were intraperitoneally injected with tilianin at doses of 3, 10, 30 mg/kg. H9c2 cells were treated with tilianin at concentrations of 10, 30, and 50 μg/mL. Echocardiography, TTC staining and TUNEL staining demonstrated that tilianin remarkably improved cardiac function and mitigated myocardial damage in MIRI rats. Additionally, notable inflammatory response reduction by tilianin was evidenced by subsequent hematatoxylin-eosin (HE) staining, inflammatory cytokines assay, and quantitative proteomics. Further western blotting analysis and immunofluorescence staining showed tilianin decreased the levels of TLR4, p-NF-κB, NLRP3, and ASC in MIRI rats and H9c2 cells exposed to OGD/R, alongside a significant reduction in cleaved gasdermin D, mature IL-1β and IL-18. Molecular docking, cellular thermal shift assay (CETSA) and co-immunoprecipitation (co-IP) assay revealed that tilianin impeded the interaction between NLRP3 and NEK7. Taken together, tilianin protects cardiomyocytes from MIRI by suppressing NLRP3 inflammasome through the inhibition of the TLR4/NF-κB signaling pathway and the disruption of the NEK7/NLRP3 interface. These findings underscore the potential of tilianin as a promising therapeutic candidate for MIRI.
田基黄苷是一种从[植物名称]中提取的黄酮类化合物，因其具有多种生物学功能而受到认可，尤其是在减轻心肌缺血再灌注损伤（MIRI）方面。有充分证据表明，NLRP3炎性小体对MIRI的发展具有重大影响。在本研究中，采用结扎并随后松开左冠状动脉前降支（LAD）的大鼠以及经历氧糖剥夺/复氧（OGD/R）的H9c2心肌细胞，来研究田基黄苷对NLRP3炎性小体的影响及其抗MIRI机制。再灌注时，给大鼠腹腔注射3、10、30mg/kg剂量的田基黄苷。用10、30和50μg/mL浓度的田基黄苷处理H9c2细胞。超声心动图、TTC染色和TUNEL染色表明，田基黄苷显著改善了MIRI大鼠的心脏功能并减轻了心肌损伤。此外，随后的苏木精-伊红（HE）染色、炎性细胞因子检测和定量蛋白质组学证明田基黄苷显著减轻了炎症反应。进一步的蛋白质印迹分析和免疫荧光染色显示，田基黄苷降低了MIRI大鼠和暴露于OGD/R的H9c2细胞中TLR4、p-NF-κB、NLRP3和ASC的水平，同时裂解的gasdermin D、成熟的IL-1β和IL-18显著减少。分子对接、细胞热位移分析（CETSA）和免疫共沉淀（co-IP）分析表明，田基黄苷阻碍了NLRP3与NEK7之间的相互作用。综上所述，田基黄苷通过抑制TLR4/NF-κB信号通路和破坏NEK7/NLRP3界面来抑制NLRP3炎性小体，从而保护心肌细胞免受MIRI损伤。这些发现强调了田基黄苷作为MIRI有前景的治疗候选药物的潜力。
