针灸和草药（英文）2026，Vol.6Issue(1)：56-72,17.DOI:10.1097/HM9.0000000000000181
冬凌草甲素通过与HMGB1共价结合抑制HMGB1/TLR4/MyD88/NF-κB信号通路减轻脑缺血再灌注损伤
Oridonin mitigates cerebral ischemic-reperfusion injury via covalent binding to HMGB1 and inhibiting the HMGB1/TLR4/MyD88/NF-κB signaling pathway
摘要
Abstract
Objective:Ischemic stroke(IS)is a leading cause of mortality and disability worldwide,and effective pharmacological treatments are limited.Oridonin(Ori)has demonstrated neuroprotective potential in IS;however,its underlying mechanisms are still poorly understood. Methods:In vitro,oxygen-glucose deprivation/reperfusion(OGD/R)models were established using mouse neuroblastoma Neuro-2a cells and primary cortical neurons.In vivo,a transient middle cerebral artery occlusion(tMCAO)model was induced in male C57BL/6J mice to simulate cerebral ischemic-reperfusion(I/R)injury.The key targets of Ori were identified using activity-based protein profiling(ABPP).The binding affinity between Ori and its target protein was validated using multiple approaches,including cellular thermal shift assay(CETSA),molecular docking,and biolayer interferometry(BLI). Results:Ori significantly suppressed the expression of inflammatory cytokines in tMCAO-and OGD/R-treated neuronal cells.Target identification revealed that high-mobility group box 1(HMGB1)protein is the key mediator of the protective effects of Ori against cerebral I/R injury.Mechanistically,Ori covalently binds to cysteine(Cys)106 of HMGB1,reducing its secretion and proinflammatory activity.Additionally,Ori downregulated cytoplasmic HMGB1 levels and the expression of TLR4 and MyD88,as well as the p-p65/p65 ratio in both OGD/R and tMCAO models.Notably,the HMGB1 inhibitor NecroX-7 conferred protection against OGD/R-induced neuronal injury and tMCAO-induced brain damage in mice,which could not be further modulated by Ori treatment. Conclusions:Our findings demonstrate that Ori confers neuroprotection against brain I/R injury by covalently binding to HMGB1 at Cys106 via its reactive carbon-carbon double bonds,thereby eliminating the proinflammatory activity of HMGB1.This molecular interaction reduces HMGB1 secretion and inhibits the downstream HMGB1/TLR4/MyD88/NF-κB signaling pathway,ultimately attenuating neuroinflammation and ischemic damage.关键词
脑缺血再灌注损伤/冬凌草甲素/HMGB1/靶点发现/化学蛋白质组学Key words
Cerebral ischemic-reperfusion injury/Chemical proteomics/HMGB1/Oridonin/Target discovery引用本文复制引用
刘丹丹,柴欣,邢佳乐,唐欢,邱崇,王培利,马昂,王继刚,张书杰,夏斐,史巧莉,赵辉,Yinkwan Wong,孟雨晴,刘艳青,朱永平..冬凌草甲素通过与HMGB1共价结合抑制HMGB1/TLR4/MyD88/NF-κB信号通路减轻脑缺血再灌注损伤[J].针灸和草药（英文）,2026,6(1):56-72,17.基金项目
The project was financially supported by the National Natural Science Foundation of China(82204672),Scientific and technological innovation project of China Academy of Chinese Medical Sciences(CI2023E002,CI2023E005TS05,CI2023E005TS08),and the Fundamental Research Funds for the Central Public Welfare Research Institutes(ZZ15-YQ-063,ZZ15-YQ-064,ZZ14-YQ-050,and ZZ17-ND-10-10). （82204672）
