Experimental Research Center, China Academy of Chinese Medical Sciences, Beijing Key Laboratory of Traditional Chinese Medicine Basic Research on Prevention and Treatment of Major Disease, Beijing, 100700, China.
Experimental Research Center, China Academy of Chinese Medical Sciences, Beijing Key Laboratory of Traditional Chinese Medicine Basic Research on Prevention and Treatment of Major Disease, Beijing, 100700, China; Henan University of Chinese Medicine, Henan, 450046, China.
J Ethnopharmacol. 2024 Mar 1;321:117438. doi: 10.1016/j.jep.2023.117438. Epub 2023 Nov 18.
Angong Niuhuang Wan (AGNHW) is a prescription from traditional Chinese medicine (TCM) that has been used for centuries to treat ischemic stroke (IS) and hemorrhagic stroke (HS). According to a recent study, targeting ferroptosis might be effective in the management of IS and HS. However, the ferroptosis-related effects and mechanisms of AGNHW have not yet been reported.
This research examines the anti-ferroptosis mechanisms of AGNHW in the treatment of IS and HS.
A system pharmacological approach including in vivo experiment, UHPLC-Q-Orbitrap HRMS, network pharmacology, molecular docking, microscale thermophoresis, and in vitro experiment was utilized to study the anti-ferroptosis mechanisms of AGNHW against IS and HS.
In vivo experiments indicated that AGNHW enhanced nerve function, decreased cerebral infarct volume, ameliorated histological brain injuries, improved the structural integrity of the blood-brain barrier, ameliorated the mitochondrial dysfunction and morphology disruption, and inhibits ROS, LPO and Fe accumulations in IS and HS rats. Using UHPLC-Q-Orbitrap HRMS, the key ingredients of AGNHW-containing serum were identified as bilirubin, berberine, baicalin, and wogonoside. According to the network pharmacology analyses, AGNHW could inhibit ferroptosis by modulating the PPAR and PI3K/AKT signaling pathways. The core targets are PPARγ, AKT, and GPX4. Molecular docking and microscale thermophoresis experiments further revealed that the key ingredients have strong interactions with ferroptosis-regulating core proteins. Moreover, in vitro experiment results showed that AGNHW alleviated ferroptosis injury induced by erastin in PC12 cells, increased cell viability, reduced the LPO and Fe levels, and up-regulated mRNA expressions of PPARγ, AKT, and GPX4. AGNHW also up-regulated protein expressions of PPARγ, p-AKT/AKT, and GPX4 in IS and HS rats.
AGNHW attenuated ferroptosis in treating IS and HS by targeting the PPARγ/AKT/GPX4 pathway. This work reveals AGNHW's anti-ferroptosis mechanism against IS and HS, but it also develops an integrated approach to demonstrate the common characteristics of drugs in treating different diseases.
安宫牛黄丸（AGNHW）是一种中药方剂，已被使用数百年用于治疗缺血性中风（IS）和出血性中风（HS）。最近的一项研究表明，针对铁死亡可能是治疗 IS 和 HS 的有效方法。然而，AGNHW 治疗 IS 和 HS 的铁死亡相关作用和机制尚未报道。
本研究探讨 AGNHW 治疗 IS 和 HS 的抗铁死亡机制。
采用系统药理学方法，包括体内实验、UHPLC-Q-Orbitrap HRMS、网络药理学、分子对接、微量热泳动和体外实验，研究 AGNHW 治疗 IS 和 HS 的抗铁死亡机制。
体内实验表明，AGNHW 可改善神经功能，降低脑梗死体积，改善脑组织损伤，改善血脑屏障结构完整性，改善线粒体功能障碍和形态破坏，抑制 IS 和 HS 大鼠 ROS、LPO 和 Fe 蓄积。通过 UHPLC-Q-Orbitrap HRMS，鉴定出 AGNHW 含血清的关键成分有胆红素、小檗碱、黄芩苷和汉黄芩苷。通过网络药理学分析，AGNHW 可通过调节 PPAR 和 PI3K/AKT 信号通路抑制铁死亡。核心靶点是 PPARγ、AKT 和 GPX4。分子对接和微量热泳动实验进一步表明，关键成分与铁死亡调节核心蛋白具有较强的相互作用。此外，体外实验结果表明，AGNHW 可减轻 erastin 诱导的 PC12 细胞铁死亡损伤，提高细胞活力，降低 LPO 和 Fe 水平，上调 PPARγ、AKT 和 GPX4 的 mRNA 表达。AGNHW 还上调了 IS 和 HS 大鼠中 PPARγ、p-AKT/AKT 和 GPX4 的蛋白表达。
AGNHW 通过靶向 PPARγ/AKT/GPX4 通路减轻 IS 和 HS 中的铁死亡。本研究揭示了 AGNHW 治疗 IS 和 HS 的抗铁死亡机制，同时也开发了一种综合方法来展示药物治疗不同疾病的共同特征。
