Mechanism of Action of Xiaoyao San in Treatment of Ischemic Stroke is Related to Anti-Apoptosis and Activation of PI3K/Akt Pathway

Yue Xu,1,* Weiyin Chen,1,* Zeran Chen,2 Mengyuan Huang,1 Fang Yang,1 Yang Zhang1 1Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu, 610072, People’s Republic of China; 2School of Medical and Life Sciences, Chengdu University of Traditional Chinese Medicine, Chengdu Sichuan, 610041, People’s Republic of China*These authors contributed equally to this workCorrespondence: Fang YangHospital of Chengdu University of Traditional Chinese Medicine, No. 39 Shi-er-qiao Road, Chengdu, 610075, People’s Republic of ChinaTel/Fax +86 28- 87783481Email yfgreens2013@163.comYang ZhangHospital of Chengdu University of Traditional Chinese Medicine, Chengdu, 610072, People’s Republic of ChinaEmail zhangyang@cdutcm.edu.cnObjective: The traditional Chinese medicine (TCM) formulation Xiaoyao San (XYS) has a good clinical effect in treating ischemic stroke (IS). We explored the mechanism and material basis of XYS in IS treatment.Methods: Network pharmacology was used to construct a network of XYS components and IS targets. R software was used to analyze the biological process and pathway analysis of the targets of XYS in IS treatment. In vitro, a model of apoptosis of PC12 cells induced by oxygen-glucose deprivation/reperfusion (OGD/R) was established to evaluate the neuroprotective effect of XYS and its influence on the expression of apoptotic protein-related genes. The affinity between the potentially active compounds in XYS and apoptotic proteins was evaluated by molecular docking.Results: XYS was shown to have 136 chemical components that exert potential anti-IS activity by acting on 175 proteins. Bioinformatics analysis showed that apoptosis and the phosphoinositide 3-kinase/protein kinase B (PI3K-Akt) signaling pathway were the main signaling pathways of XYS. In vitro experiments showed that XYS could improve the effect of OGD/R on PC12-cell activity (EC50 = 0.43 mg/mL) and inhibit apoptosis. The main mechanisms were related to the improvement of oxidative stress and regulation of apoptosis-related gene expression. Molecular docking showed that C22, C102 and other components in XYS had a strong affinity with apoptosis-related proteins.Conclusion: Network pharmacology, in vitro experiments, and molecular docking were used, for the first time, to study the material basis and molecular mechanism of XYS in IS treatment from the perspective of multiple targets and multiple pathways. We provided a new approach for the future study of TCM formulations in the treatment of complex diseases.Keywords: apoptosis, neuroprotection, biological function, pharmacological mechanism