Xu Libin, Li Siyu, Qi Jiaxin, Mi Yan, Zhang Ying, Yang Yuxin, Wang Yingjie, Zhou Di, Li Ning, Hou Yue
Key Laboratory of Bioresource Research and Development of Liaoning Province, College of Life and Health Sciences, National Frontiers Science Center for Industrial Intelligence and Systems Optimization, Key Laboratory of Data Analytics and Optimization for Smart Industry, Ministry of Education, Northeastern University, Shenyang, PR China.
School of Traditional Chinese Materia Medica, Key Laboratory of Innovative Traditional Chinese Medicine for Major Chronic Diseases of Liaoning province, Key Laboratory for TCM Material Basis Study and Innovative Drug Development of Shenyang City, Shenyang Pharmaceutical University, Shenyang 110016, PR China.
Phytomedicine. 2025 Jan;136:156253. doi: 10.1016/j.phymed.2024.156253. Epub 2024 Nov 25.
The significance of pyroptosis as an inflammatory mode of death in ischemic stroke (IS) has attracted much attention in recent years. Effusol is a dihydrophenanthrene component extracted from Juncus effusus L.. Previous studies have found that Juncus effusus L. has a good inhibitory effect against microglia activation. However, it is not clear whether effusol inhibits microglia over-activation and attenuates its mediated microglia pyroptosis in the treatment of IS.
The aim is to examine how effusol influences the initiation and activation stages of pyroptosis, as well as the NLRP3 inflammasome, resulting from microglial over-activation triggered post-IS.
This study investigated the impact of effusol on neurological severity and edema to assess its neuroprotective effects in IS. Mechanistically, immunofluorescence and western blotting were applied to explore the initiation and activation of the NLRP3 inflammasome. Finally, we employed the NLRP3 specific inhibitor, molecular docking, drug affinity responsive target stability (DARTS), and cellular thermal shift assay (CETSA) to further explore the underlying targets of effusol.
Effusol mitigated IS-induced damage and downregulated the expression of inflammatory factors at the mRNA level, the protein levels of toll-like receptor 4 (TLR4), nuclear transcription factor NF-κB p65, and key components of the NLRP3 inflammasome. Effusol also mitigated mitochondrial damage by increasing ATP levels and decreasing mitochondrial membrane potential. Importantly, effusol targets NLRP3 protein to inhibit pyroptosis, thereby suppressing the hyperactivation of NLRP3 inflammasome.
Effusol may be protective against IS by targeting NLRP3 proteins to inhibit NLRP3 inflammasome activation-mediated pyroptosis. This finding provides a theoretical basis and a prospective drug candidate for the treatment of effusol in IS.
近年来，焦亡作为缺血性中风（IS）中一种炎症性死亡方式的重要性备受关注。艾黄素是从灯心草中提取的一种二氢菲成分。先前的研究发现，灯心草对小胶质细胞活化具有良好的抑制作用。然而，尚不清楚艾黄素在IS治疗中是否能抑制小胶质细胞过度活化并减轻其介导的小胶质细胞焦亡。
目的是研究艾黄素如何影响由IS后触发的小胶质细胞过度活化所导致的焦亡起始和激活阶段，以及NLRP3炎性小体。
本研究调查了艾黄素对神经功能严重程度和水肿的影响，以评估其在IS中的神经保护作用。从机制上讲，应用免疫荧光和蛋白质印迹法来探究NLRP3炎性小体的起始和激活。最后，我们使用NLRP3特异性抑制剂、分子对接、药物亲和力响应靶点稳定性（DARTS）和细胞热位移分析（CETSA）来进一步探究艾黄素的潜在靶点。
艾黄素减轻了IS诱导的损伤，并在mRNA水平下调了炎症因子的表达，以及Toll样受体4（TLR4）、核转录因子NF-κB p65和NLRP3炎性小体关键成分的蛋白质水平。艾黄素还通过增加ATP水平和降低线粒体膜电位减轻了线粒体损伤。重要的是，艾黄素靶向NLRP3蛋白以抑制焦亡，从而抑制NLRP3炎性小体的过度活化。
艾黄素可能通过靶向NLRP3蛋白抑制NLRP3炎性小体激活介导的焦亡而对IS具有保护作用。这一发现为艾黄素在IS治疗中的应用提供了理论依据和一种有前景的候选药物。
