Li Yanling, Hu Zhongji, Xie Linli, Xiong Tingting, Zhang Yanyan, Bai Yang, Ding Huang, Huang Xiaoping, Liu Xiaodan, Deng Changqing
Hunan University of Chinese Medicine, Changsha, 410208, China; Hunan Province Key Laboratory of Cerebrovascular Disease Prevention and Treatment of Integrated Chinese Medicine and Western Medicine, Changsha, 410208, China.
Hunan University of Chinese Medicine, Changsha, 410208, China.
J Ethnopharmacol. 2025 Jan 31;340:119246. doi: 10.1016/j.jep.2024.119246. Epub 2024 Dec 15.
Buyang Huanwu Decoction (BYHWD) exerts its anti-cerebral ischemia effects through multiple pathways and targets, although its specific mechanisms remain unclear.
Ultra-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UPLC-QTOF-MS) metabolomics and other methods were employed to investigate the role of BYHWD in inhibiting neuronal apoptosis following cerebral ischemia-reperfusion by modulating the RhoA/Rock2 pathway.
A rat model of exhaustion swimming combined with middle cerebral artery occlusion (ES + I/R) was established to evaluate the intervention effects of Buyang Huanwu Decoction on cerebral ischemia-reperfusion. This was assessed using neurological function scores, Qi deficiency and blood stasis syndrome scores, HE staining, Nissl staining and TT staining. Differential metabolites and metabolic pathways associated with cerebral ischemia-reperfusion were identified using UPLC-QTOF-MS metabolomics, with key differential metabolites validated through ELISA. Molecular docking techniques were employed to predict interactions between the key differential metabolite, hippuric acid, and its primary downstream pathways. Finally, the levels of neurocellular apoptosis, as well as key molecules in the RhoA/Rock2 signaling pathway and the mitochondrial apoptosis pathway, were measured.
The results indicated that the primary differential metabolites associated with BYHWD's protective effects against ischemia-reperfusion injury were hippuric acid, lysophosphatidic acid, and lysophosphatidylethanolamine, with the main metabolic pathway being phenylalanine metabolism. Molecular docking studies demonstrated that malonic acid exhibited a strong affinity for proteins related to the RhoA/Rock2 signaling pathway and the mitochondrial apoptosis pathway.Furthermore, we found that BYHWD treatment significantly decreased the apoptosis rate of cells following cerebral ischemia-reperfusion and inhibited the expression of key molecules in both the RhoA/Rock2 signaling pathway and the mitochondrial apoptosis pathway in brain tissue.
BYHWD ameliorated brain tissue injury after cerebral ischemia/reperfusion in rats with qi deficiency and blood stasis. The underlying mechanism may involve BYHWD's inhibition of the RhoA/Rock2 signaling pathway activation through modulation of the phenylalanine metabolism pathway, thereby reducing neuronal apoptosis mediated by the mitochondrial apoptosis pathway.
补阳还五汤（BYHWD）通过多种途径和靶点发挥抗脑缺血作用，但其具体机制尚不清楚。
采用超高效液相色谱-四极杆飞行时间串联质谱（UPLC-QTOF-MS）代谢组学等方法，研究补阳还五汤通过调节RhoA/Rock2通路在抑制脑缺血再灌注后神经元凋亡中的作用。
建立力竭游泳联合大脑中动脉闭塞（ES + I/R）大鼠模型，以评价补阳还五汤对脑缺血再灌注的干预效果。采用神经功能评分、气虚血瘀证评分、HE染色、尼氏染色和TT染色进行评估。运用UPLC-QTOF-MS代谢组学鉴定与脑缺血再灌注相关的差异代谢物和代谢途径，并通过ELISA验证关键差异代谢物。采用分子对接技术预测关键差异代谢物马尿酸与其主要下游途径之间的相互作用。最后，检测神经细胞凋亡水平以及RhoA/Rock2信号通路和线粒体凋亡通路中的关键分子。
结果表明，与补阳还五汤对缺血再灌注损伤保护作用相关的主要差异代谢物为马尿酸、溶血磷脂酸和溶血磷脂酰乙醇胺，主要代谢途径为苯丙氨酸代谢。分子对接研究表明，丙二酸对RhoA/Rock2信号通路和线粒体凋亡通路相关蛋白具有较强亲和力。此外，我们发现补阳还五汤治疗可显著降低脑缺血再灌注后细胞凋亡率，并抑制脑组织中RhoA/Rock2信号通路和线粒体凋亡通路关键分子的表达。
补阳还五汤改善了气虚血瘀型大鼠脑缺血/再灌注后的脑组织损伤。其潜在机制可能涉及补阳还五汤通过调节苯丙氨酸代谢途径抑制RhoA/Rock2信号通路激活，从而减少线粒体凋亡途径介导的神经元凋亡。
