基于PGC-1α/ERRα调控内皮细胞焦亡研究杜仲抗脑缺血的活性成分及作用机制

中药杜仲“补肝肾、强筋骨”，据“肾主骨生髓通与脑”的传统中医理论，临床上多用于治疗肾虚相关疾病如呆症、痿症、中风、眩晕等。课题组前期研究发现杜仲提取物具有显著抗脑缺血活性，属于中医“中风”和“眩晕”范畴，但其体内代谢特征和抗脑缺血有效成分及机制并不明确。本项目拟在前期研究基础上，解析杜仲的体内代谢规律和组织分布特征；建立缺血性脑微血管内皮细胞焦亡损伤模型，以杜仲抗脑缺血内皮焦亡有效成分为研究对象，利用化学生物学技术揭示其直接作用靶标；结合内皮特异靶标敲除动物模型，利用代谢组和转录组等生物信息分析和验证实验，明确杜仲抗脑缺血内皮焦亡的作用蛋白和调控基因网络，阐明其在体内外脑缺血模型中通过调控PGC-1α/ERRα/TNFAIP3信号通路抑制脑微血管内皮细胞焦亡进而保护血脑屏障改善脑缺血损伤的的分子机理。本项目将为杜仲治疗缺血性脑卒中的药效物质和新靶标发现及作用机制研究提供理论依据和数据支撑。

The traditional Chinese medicine (TCM), Eucommia ulmoides Oliver has the function of invigorating liver and kidney and strengthening muscles and bones. According to the traditional Chinese medicine theory of “Kidney governing bone, generating marrow through the brain”, it is mostly used to treat kidney deficiency-related diseases, such as dementia, impotence, stroke, vertigo and so on. Previous studies of our group have found that Eucommia ulmoides extract has significant anti-cerebral ischemic activity, belonging to the category of “stroke” and “vertigo” in TCM, but its metabolic characteristics and anti-cerebral ischemic active ingredients with mechanisms are not clear. On the basis of our previous studies, this project aims to analyze the metabolic regularity and tissue distribution characteristics of Eucommia ulmoides Oliver; discover the active ingredients of Eucommia ulmoides Oliver with anti-cerebral ischemic endothelial pyroptosis in established pyroptosis model of ischemic brain microvascular endothelial cells, and elucidate its direct protein target utilizing chemical biology technology; combining with endothelium-specific protein target gene knockout animal model, using bioinformatics analysis and validation experiments including metabonomics and transcriptome, clarify the acting proteins and regulatory gene network of Eucommia ulmoides Oliver against cerebral ischemic endothelial pyroptosis, further elucidating its molecular mechanisms for improving cerebral ischemic injury by regulating PGC-1α/ERRα/TNFAIP3 signaling pathway to inhibit brain microvascular endothelial pyroptosis and protect blood-brain barrier integrity in vitro and in vivo. This project will provide theoretical basis and data support for the discovery of effective substances and new targets of Eucommia ulmoides Oliver in the treatment of brain ischemic stroke and the study of mechanism of action.