Xiaoxuming decoction enhanced neuroprotection after cerebral ischemia/reperfusion via the JAK2/STAT3 signaling pathway based on UPLC/HRMS, network pharmacology and experimental validation

Manman Wang; Yong Zhang; Xueqin Fu; Xuhuan Zou; Jun Xiang; Rui Lan

doi:10.1016/j.jep.2024.119279

Xiaoxuming decoction enhanced neuroprotection after cerebral ischemia/reperfusion via the JAK2/STAT3 signaling pathway based on UPLC/HRMS, network pharmacology and experimental validation

J Ethnopharmacol. 2024 Dec 24:340:119279. doi: 10.1016/j.jep.2024.119279. Online ahead of print.

Authors

Manman Wang¹, Yong Zhang², Xueqin Fu¹, Xuhuan Zou¹, Jun Xiang³, Rui Lan⁴

Affiliations

¹ The First Clinical Medical school of Henan University of Chinese Medicine, Henan, 450000, China.
² The Third Affiliated Hospital of Zhengzhou University, Henan, 450000, China.
³ Department of Integrative Medicine, Zhongshan Hospital, Fudan University, 200032, Shanghai, China.
⁴ Encephalopathy Hospital, The First Affiliated Hospital of Henan University of Chinese Medicine, Henan, 450000, China. Electronic address: lanrui@hactcm.edu.cn.

PMID: 39725365
DOI: 10.1016/j.jep.2024.119279

Abstract

Ethnopharmacological relevance: Xiao-xu-ming decoction (XXMD), a prominent traditional Chinese medicinal formula historically revered for stroke treatment, demonstrates pronounced efficacy in ameliorating ischemic stroke injury.

Aim of the study: This study aims to investigate the effects and mechanisms of XXMD on neuroprotection subsequent to cerebral ischemia/reperfusion in vivo and in vitro.

Materials and methods: Neurobehavioral test, TTC staining, HE staining and nissl staining were used to examine the neuroprotective effect of XXMD on cerebral ischemia-reperfusion injury induced by middle cerebral artery occlusion (MCAO) in rats. Additionally, we assessed cell viability and injury with CCK8 and lactate dehydrogenase (LDH) assays. The changes in neuronal ultra-structure were observed after oxygen-glucose deprivation and reoxygenation (OGD/R) by transmission electron microscopy (TEM). Network analysis combined with ultrahighperformance liquid chromatography-high resolution mass spectrometry (UPLC-HRMS) predicted the mechanism of XXMD on ischemic stroke injury. Furthermore, the expression of neuroplasticity-related proteins neurofilament 200 (NF200), microtubule-associated protein 2 (MAP2), postsynaptic density protein 95 (PSD95), synaptophysin (SYN), phosphorylated Janus kinase2 (p-JAK2), and phosphorylated signal transduction and activator of transcription 3 (p-STAT3) was evaluated by immunofluorescence staining and Western blot analyses.

Results: XXMD significantly improved Ethology, infarct area and pathological changes after MCAO and reperfusion, reducing morphological and ultrastructural alterations and decreased cell viability in HT22 cells induced by OGD/R. Network pharmacology showed that 1153 compounds of XXMD were matched. The Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis demonstrated that XXMD treated ischemia stroke mainly regulating inflammation reaction-related signaling pathways, atherosclerosish-related signaling pathways. Molecular docking results showed that TP53, AKT1, STAT3, and IL6 are closely bound to the corresponding active ingredients. XXMD treatment significantly reversed the above alternations. XXMD or AG490 up-regulated the expression of neuroplasticity-associated proteins, and reduced phosphorylation of JAK2, STAT3 expression following OGD/R.

Conclusion: XXMD exerts neuroprotective effects by promoting neural plasticity via regulating the JAK2/STAT3 pathway, indicating a promising alternative therapeutic strategy for ischemic stroke.

Keywords: Cerebral ischemia-reperfusion injury; JAK2/STAT3 pathway; Neural plasticity; Oxygen-glucose deprivation/reoxygenation; Xiaoxuming decoction.