毛蕊异黄酮靶向tRFi-Ser-25a/SOS1轴介导脑出血后神经保护作用的机制研究

补阳还五汤改善脑出血（ICH）后神经损伤疗效确切。我们前期发现转运RNA来源小RNA（tsRNA）是补阳还五汤治疗ICH的新型靶点，但精准机制尚未阐明。基于此，申请人采用现代网络药理学和成分分析技术发现毛蕊异黄酮是补阳还五汤的药效物质基础，发挥母方改善ICH后神经功能和调节tRFi-Ser-25a的作用。深入生物信息学分析和预实验结果提示tRFi-Ser-25a可靶向调控SOS1，并调节RAS/ERK信号通路。据此，我们提出假说：毛蕊异黄酮通过靶向tRFi-Ser-25a/SOS1轴，抑制RAS/ERK信号通路减轻神经损伤，发挥神经保护作用。本项目拟通过转染技术、双荧光素酶报告基因、RNA结合蛋白免疫沉淀等方法，在体内和体外水平，深入探讨毛蕊异黄酮以tRFi-Ser-25a为靶点改善ICH后神经元损伤的作用及分子机制，为毛蕊异黄酮治疗ICH提供新的理论依据和新视角。

Buyang Huanwu decoction is effective in improving nerve injury after intracerebral hemorrhage (ICH). We have previously found that transfer RNA-derived small RNAs (tsRNAs) may be a new drug target for Buyang Huanwu decoction in treating ICH. However, the precise mechanism has not been clarified. Based on this, we found that calycosin was the pharmacodynamic material basis of Buyang Huanwu decoction through modern network pharmacology and component analysis techniques. Moreover, calycosin played the role of Buyang Huanwu decoction in improving neurological function after ICH and regulating tRFi-Ser-25a. Further bioinformatics analysis and pre-experimental results suggested that tRFi-Ser-25a could target SOS1 and regulate RAS/ERK signaling pathway. Accordingly, we propose a hypothesis that calycosin can target tRFi-Ser-25a/SOS1 axis and inhibit RAS/ERK signaling pathway to alleviate nerve injury and exert neuroprotective effects. To confirm this scientific hypothesis, this project aims to investigate the effects and molecular mechanisms of calycosin-mediated neuroprotection by targeting tRFi-Ser-25a in vivo and in vitro, through transfection, Dual-luciferase reporter gene assay, RNA-binding protein immunoprecipitation and other methods. Our work will provide the new evidence and novel perspective for ICH treated with calycosin.