Electroacupuncture Inhibits Neural Ferroptosis in Rat Model of Traumatic Brain Injury via Activating System Xc-/GSH/GPX4 Axis

Na Li; Ruihui Wang; Xia Ai; Jie Guo; Yuwang Bai; Xinrong Guo; Rongchao Zhang; Xu Du; Jingxuan Chen; Hua Li

doi:10.2174/0115672026297775240405073502

Electroacupuncture Inhibits Neural Ferroptosis in Rat Model of Traumatic Brain Injury via Activating System Xc-/GSH/GPX4 Axis

Curr Neurovasc Res. 2024 Apr 15. doi: 10.2174/0115672026297775240405073502. Online ahead of print.

Authors

Na Li^{1

2}, Ruihui Wang², Xia Ai², Jie Guo², Yuwang Bai³, Xinrong Guo², Rongchao Zhang², Xu Du², Jingxuan Chen², Hua Li²

Affiliations

¹ School of Acupuncture-Tuina, Chengdu University of Traditional Chinese Medicine, Chengdu, Sichuan, 610075, China.
² School of Acupuncture-Tuina, Shaanxi University of Traditional Chinese Medicine, Xi'an, Shaanxi, 712046, China.
³ Xi'an Hospital of Traditional Chinese Medicine, Xi'an, Shaanxi, 710001, China.

PMID: 38629369
DOI: 10.2174/0115672026297775240405073502

Abstract

Background: Ferroptosis is an iron-dependent regulating programmed cell death discovered recently that has been receiving much attention in traumatic brain injury (TBI). xCT, a major functional subunit of Cystine/glutamic acid reverse transporter (System Xc-), promotes cystine intake and glutathione biosynthesis, thereby protecting against oxidative stress and ferroptosis.

Objective: The intention of this research was to verify the hypothesis that electroacupuncture (EA) exerted an anti-ferroptosis effect via an increase in the expression of xCT and activation of the System Xc-/GSH/GPX4 axis in cortical neurons of TBI rats.

Methods: After the TBI rat model was prepared, animals received EA treatment at GV20, GV26, ST36 and PC6, for 15min. The xCT inhibitor Sulfasalazine (SSZ) was administered 2h prior to model being prepared. The degree of neurological impairment was evaluated by means of TUNEL staining and the modified neurological severity score (mNSS). Specific indicators of ferroptosis (Ultrastructure of mitochondria, Iron and ROS) were detected by transmission electron microscopy (TEM), Prussian blue staining (Perls stain) and flow cytometry (FCM), respectively. GSH synthesis and metabolism-related factors in the content of the cerebral cortex were detected by an assay kit. Real-time quantitative PCR (RT-QPCR), Western blot (WB), and immunofluorescence (IF) were used for detecting the expression of System Xc-/GSH/GPX4 axisrelated proteins in injured cerebral cortex tissues.

Results: EA successfully relieved nerve damage within 7 days after TBI, significantly inhibited neuronal ferroptosis, upregulated the expression of xCT and System Xc-/GSH/GPX4 axis forward protein and promoted glutathione (GSH) synthesis and metabolism in the injured area of the cerebral cortex. However, aggravation of nerve damage and increased ferroptosis effect were found in TBI rats injected with xCT inhibitors.

Conclusions: EA inhibits neuronal ferroptosis by up-regulated xCT expression and by activating System Xc-/GSH/GPX4 axis after TBI, confirming the relevant theories regarding the EA effect in treating TBI and providing theoretical support for clinical practice.

Keywords: Traumatic brain injury; electroacupuncture; ferroptosis; system Xc− /GSH/GPX4 axis..