THSG alleviates cerebral ischemia/reperfusion injury via the GluN2B-CaMKII-ERK1/2 pathway

Tonghe Liu; Jiayi Shi; Dahua Wu; Dandan Li; Yuhong Wang; Jian Liu; Pan Meng; Lijuan Hu; Chaojun Fu; Zhigang Mei; Jinwen Ge; Xiuli Zhang

doi:10.1016/j.phymed.2024.155595

THSG alleviates cerebral ischemia/reperfusion injury via the GluN2B-CaMKII-ERK1/2 pathway

Phytomedicine. 2024 Apr 6:129:155595. doi: 10.1016/j.phymed.2024.155595. Online ahead of print.

Authors

Tonghe Liu¹, Jiayi Shi², Dahua Wu³, Dandan Li⁴, Yuhong Wang², Jian Liu⁵, Pan Meng⁴, Lijuan Hu⁴, Chaojun Fu⁴, Zhigang Mei⁶, Jinwen Ge⁷, Xiuli Zhang⁸

Affiliations

¹ Institute of Innovation and Applied Research, Hunan University of Chinese Medicine, 300 Bachelor Road, Changsha 410208, China; Chinese Academy of Medical Sciences, Institute of Medicinal Plant Development, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing 100193, China.
² Institute of Innovation and Applied Research, Hunan University of Chinese Medicine, 300 Bachelor Road, Changsha 410208, China.
³ Department of Neurology, Hunan University of Chinese Medicine Integrated Chinese Medicine Affiliated Hospital, Changsha 410208, China.
⁴ Hunan University of Chinese Medicine, Changsha 410208, China.
⁵ The First Hospital, Hunan University of Chinese Medicine, Changsha 410208, China.
⁶ The Key Laboratory of Hunan Province for Integrated Traditional Chinese and Western Medicine on Prevention and Treatment of Cardio-Cerebral Diseases, Hunan University of Chinese Medicine, 300 Bachelor Road, Changsha 410208, China. Electronic address: meizhigang@hnucm.edu.cn.
⁷ The Key Laboratory of Hunan Province for Integrated Traditional Chinese and Western Medicine on Prevention and Treatment of Cardio-Cerebral Diseases, Hunan University of Chinese Medicine, 300 Bachelor Road, Changsha 410208, China. Electronic address: 001267@hnucm.edu.cn.
⁸ Institute of Innovation and Applied Research, Hunan University of Chinese Medicine, 300 Bachelor Road, Changsha 410208, China. Electronic address: 004115@hnucm.edu.cn.

PMID: 38677275
DOI: 10.1016/j.phymed.2024.155595

Abstract

Background: The potential therapeutic targeting of PINK1-PARK2-mediated mitophagy against cerebral ischemia/reperfusion (CI/R) injury involves the pathophysiological processes of neurovascular unit (NVU) and is closely associated with N-methyl-D-aspartate receptors (NMDARs) commonly expressed in NVU. 2,3,5,4'-Tetrahydroxy-stilbene-2-O-β-D-glucoside (THSG), a compound derived from the traditional Chinese medicine Polygonum multiflorum Thunb., has demonstrated notable neuroprotective properties against CI/R injury. However, it remains unclear whether THSG exerts its protective effects through GluN2B related PINK1/ PARK2 pathway.

Purpose: This study aims to explore the pharmacological effects of THSG on alleviating CI/R injury via the GluN2B-CaMKII-ERK1/2 pathway.

Methods: THSG neuroprotection against CI/R injury was studied in transient middle cerebral artery occlusion/reversion (tMCAO/R) model rats and in oxygen and glucose deprivation/ reoxygenation (OGD/R) induced neurons. PINK1-PARK2-mediated mitophagy involvement in the protective effect of THSG was investigated in tMCAO/R rats and OGD/R-induced neurons via THSG and 3-methyladenine (3-MA) treatment. Furthermore, the beneficial role of GluN2B in reperfusion and its contribution to the THSG effect via CaMKII-ERK1/2 and PINK1-PARK2-mediated mitophagy was explored using the GluN2B-selective antagonist Ro 25-6981 both in vivo and in vitro. Finally, the interaction between THSG and GluN2B was evaluated using molecular docking.

Results: THSG significantly reduced infarct volume, neurological deficits, penumbral neuron structure, and functional damage, upregulated the inhibitory apoptotic marker Bcl-2, and suppressed the increase of pro-apoptotic proteins including cleaved caspase-3 and Bax in tMCAO/R rats. THSG (1 μM) markedly improved the neuronal survival under OGD/R conditions. Furthermore, THSG promoted PINK1 and PARK2 expression and increased mitophagosome numbers and LC3-II-LC3-I ratio both in vivo and in vitro. The effects of THSG were considerably abrogated by the mitophagy inhibitor 3-MA in OGD/R-induced neurons. Inhibiting GluN2B profoundly decreased mitophagosome numbers and OGD/R-induced neuronal viability. Specifically, inhibiting GluN2B abolished the protection of THSG against CI/R injury and reversed the upregulation of PINK1-PARK2-mediated mitophagy by THSG. Inhibiting GluN2B eliminated THSG upregulation of ERK1/2 and CaMKII phosphorylation. The molecular docking analysis results demonstrated that THSG bound to GluN2B (binding energy: -5.2 ± 0.11 kcal/mol).

Conclusions: This study validates the premise that THSG alleviates CI/R injury by promoting GluN2B expression, activating CaMKII and ERK1/2, and subsequently enhancing PINK1-PARK2-mediated mitophagy. This work enlightens the potential of THSG as a promising candidate for novel therapeutic strategies for treating ischemic stroke.

Keywords: 2,3,5,4′-tetrahydroxystilbene-2-O-β-d-glucoside (THSG); CaMKⅡ; Cerebral ischemia/ reperfusion (CI/R); ERK1/2; GluN2B; PINK1/PARK2-mediated mitophagy.