Mao Mingjiang, Shentu Chenhuan, Chen Xueao, Meng Qingling, Jiao Ziyu, Zhang Yikai, Zhu Na, Zhou Liping, Wu Yangsheng, Dai Shijie, Yuan Xiaofeng
the School of Life Sciences, Zhejiang Chinese Medical University, Hangzhou, Zhejiang, 310053, China.
the Fifth School of Clinical Medicine, Zhejiang Chinese Medical University, Hangzhou, Zhejiang, 310053, China.
J Ethnopharmacol. 2025 Sep 24;355(Pt A):120644. doi: 10.1016/j.jep.2025.120644.
Atractylodes macrocephala, traditionally recorded in classical formulas such as 'Banxia Baizhu Tianma Tang' and 'Xiaoxuming Tang', has been used in traditional medicine for conditions described as 'fēng tán zǔ luò xíng' and 'bì zǔ jīng luò xíng', which are considered related to cerebrovascular disorders. Atractylenolide Ⅲ (ATL Ⅲ), a typical sesquiterpene lactone derived from A. macrocephala, has been reported to exert neuroprotective effects through antioxidant and anti-inflammatory activities. Nevertheless, its specific role and underlying mechanisms in cerebral ischemia reperfusion injury (CIRI) remain to be clarified.
The present study was designed to test the hypothesis that ATL Ⅲ ameliorates CIRI primarily by suppressing inflammation through the PI3K/Akt/NF-κB signaling pathway, which was evaluated in both middle cerebral artery occlusion (MCAO) and oxygen-glucose deprivation/reoxygenation (OGD/R) models.
In this study, models of MCAO and OGD/R were established to explore its effect of ATL Ⅲ on CIRI. Thereafter, the therapeutic mechanism of ATL Ⅲ via transcriptomics, molecular docking, enzyme-linked immunosorbent assay (ELISA), immunohistochemistry, immunofluorescence, Western blot analysis.
ATL Ⅲ treatment significantly reduced infarct volume, neurological deficits, and pro-inflammatory cytokine release, while preserving blood-brain barrier (BBB) integrity in MCAO mice. In OGD/R-exposed PC12 cells, ATL Ⅲ attenuated oxidative stress, inhibited apoptosis, and decreased inflammatory mediator production. Transcriptomic analysis revealed several significantly enriched pathways following ATL Ⅲ treatment, among which the PI3K/Akt/NF-κB signaling pathway was prominent and therefore guided our mechanistic focus. Molecular docking supported the binding of ATL Ⅲ to key pathway proteins, and inhibition of PI3K with LY294002 attenuated the protective effects of ATL Ⅲ, confirming the central role of this pathway.
ATL Ⅲ may inhibit inflammation in CIRI, potentially through regulation of the PI3K/Akt/NF-κB signaling pathway, highlighting its promise as a candidate compound for further preclinical investigation in ischemic stroke.
白术传统上记载于“半夏白术天麻汤”和“小续命汤”等经典方剂中，在传统医学中用于治疗“风痰阻络型”和“痹阻经络型”病症，这些病症被认为与脑血管疾病有关。白术内酯Ⅲ（ATLⅢ）是一种从白术中提取的典型倍半萜内酯，据报道可通过抗氧化和抗炎活性发挥神经保护作用。然而，其在脑缺血再灌注损伤（CIRI）中的具体作用和潜在机制仍有待阐明。
本研究旨在验证ATLⅢ主要通过PI3K/Akt/NF-κB信号通路抑制炎症来改善CIRI的假设，该假设在大脑中动脉闭塞（MCAO）和氧糖剥夺/复氧（OGD/R）模型中均进行了评估。
在本研究中，建立了MCAO和OGD/R模型以探讨ATLⅢ对CIRI的影响。此后，通过转录组学、分子对接、酶联免疫吸附测定（ELISA）、免疫组织化学、免疫荧光、蛋白质印迹分析等方法研究ATLⅢ的治疗机制。
ATLⅢ治疗显著减少了MCAO小鼠的梗死体积、神经功能缺损和促炎细胞因子释放，同时保持了血脑屏障（BBB）的完整性。在OGD/R处理的PC12细胞中，ATLⅢ减轻了氧化应激，抑制了细胞凋亡，并减少了炎症介质的产生。转录组分析显示，ATLⅢ处理后有几个显著富集的通路，其中PI3K/Akt/NF-κB信号通路最为突出，因此成为我们机制研究的重点。分子对接支持ATLⅢ与关键通路蛋白的结合，用LY294002抑制PI3K可减弱ATLⅢ的保护作用，证实了该通路的核心作用。
ATLⅢ可能通过调节PI3K/Akt/NF-κB信号通路抑制CIRI中的炎症，突出了其作为缺血性中风进一步临床前研究候选化合物的潜力。
