补阳还五汤调控焦亡保护脑缺血再灌注损伤的作用机制  

Mechanism of regulating pyroptosis and protecting cerebral ischemia reperfusion injury by Buyang Huanwu Decoction

作　　者：曹卢园 郑小小 尹安康 赵琴琴 葛淑瑜 王宏伟 CAO Lu-yuan;ZHENG Xiao-xiao;YIN An-kang;ZHAO Qin-qin;GE Shu-yu;WANG Hong-wei(School of Basic Medical Sciences and Forensic Medicine,Hangzhou Medical College,Hangzhou 310013,China;Cancer Institute of Integrated Traditional Chinese and Western Medicine,Zhejiang Academy of Traditional Chinese Medicine,Hangzhou 310012,China;College of Medical Technology and Information Engineering,Zhejiang Chinese MedicineUniversity,Hangzhou310053,China;Tongde Hospital of Zhejiang Province,Hangzhou 310012,China)

机构地区：[1]杭州医学院,基础医学与法医学院,浙江杭州310013 [2]浙江省中医药研究院,中西医结合肿瘤研究所,浙江杭州310012 [3]浙江中医药大学,医学技术与信息工程学院,浙江杭州310053 [4]浙江省立同德医院,浙江杭州310012

出　　处：《药学学报》2025年第3期615-626,共12页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(82174132,82374293);浙江省医药卫生科技计划项目(2022PY044)。

摘　　要：本研究旨在探究补阳还五汤(Buyang Huanwu Decoction,BYHWD)通过抑制焦亡缓解脑缺血再灌注损伤(cerebral ischemia-reperfusion injury,CIRI)的作用机制及有效成分。首先通过网络药理学分析预测BYHWD治疗CIRI的关键成分及靶点,并通过Autodock软件和Pymol软件进行分子对接,再通过体内外实验验证BYHWD及相关有效成分的作用。体内利用小鼠大脑中动脉栓塞(middle cerebral artery occlusion,MCAO)建立模型,评估不同条件下小鼠神经功能;体外利用小鼠脑组织星形胶质细胞,建立氧糖剥夺/再灌注(oxygen-glucose deprivation/reperfusion,OGD/R)模型,并利用分子生物学实验,验证预测的关键靶点。发现BYHWD的主要成分为黄芩素、β-谷甾醇,通过分析BYHWD与CIRI疾病相关基因以及焦亡相关基因,交集的基因共20个,结合分子对接结合能,选取其中TP53和TNF关键核心靶点进行后续验证。分子实验明确BYHWD可有效减轻损伤,并降低P53的表达。上述结果提示,补阳还五汤主要活性成分黄芩素、β-谷甾醇等通过TNF和TP53等靶点抑制焦亡,发挥保护CIRI的作用。本实验已由浙江省中医药研究院实验动物福利伦理委员会批准(批准号:KTSC2020037、KTSC2023030)。This study aimed to clarify the mechanism and active components of Buyang Huanwu Decoction(BYHWD)in alleviating cerebral ischemia reperfusion injury(CIRI)by inhibiting pyroptosis.The key components and targets of BYHWD for CIRI were identified via network pharmacological analysis,followed by molecular docking performed with Autodock and Pymol software.The effects of BYHWD and its active components were validated in vivo and in vitro.A middle cerebral artery occlusion(MCAO)model was established in mouse to assess neural function alterations in mice under various conditions.Concurrently,an oxygen-glucose deprivation/reperfusion(OGD/R)model was developed utilizing mouse brain tissue astrocytes in vitro.Molecular biology experiments were used to verify the predicted key targets.We have determined that the principal components of BYHWD are baicalein andβ-sitosterol.By analyzing genes associated with CIRI pathology alongside those linked to pyroptosis,20 intersecting genes were identified.In conjunction with molecular docking binding energy assessment,TP53 and TNF emerged as pivotal core targets for subsequent validation.Molecular biology experiments confirmed that BYHWD effectively alleviates injury while reducing the expression level of P53.These findings indicate that the primary bioactive constituents of BYHWD were baicalein andβ-sitosterol.In addition,BYHWD may inhibit pyroptosis via TNF and TP53 in protecting CIRI.The experiment has been approved by the Experimental Animal Welfare Ethics Committee of Zhejiang Academy of Traditional Chinese Medicine,approval number(KTSC2020037,KTSC2023030).

关 键 词：补阳还五汤 脑缺血再灌注损伤 焦亡 网络药理学 

分 类 号：R966[医药卫生—药理学]