Salvia miltiorrhiza Bge. processed with porcine cardiac blood inhibited GLRX5-mediated ferroptosis alleviating cerebral ischemia-reperfusion injury

Shikang Zhou; Ziqi Wang; Ting Wang; Chunhua Peng; Jinyun Zhang; Chanming Liu; Jianda Xu; Yi Zhang; Li Zhang; Libo Luo; Xiaojing Yan

doi:10.1016/j.phymed.2024.155622

Salvia miltiorrhiza Bge. processed with porcine cardiac blood inhibited GLRX5-mediated ferroptosis alleviating cerebral ischemia-reperfusion injury

Phytomedicine. 2024 Apr 25:129:155622. doi: 10.1016/j.phymed.2024.155622. Online ahead of print.

Authors

Shikang Zhou¹, Ziqi Wang², Ting Wang², Chunhua Peng², Jinyun Zhang³, Chanming Liu³, Jianda Xu³, Yi Zhang⁴, Li Zhang⁵, Libo Luo⁶, Xiaojing Yan⁷

Affiliations

¹ Changzhou Key Laboratory of Human Use Experience Research & Transformation of Menghe Medical Sect, Changzhou Hospital of Chinese Medicinal Affiliated to Nanjing University of Chinese Medicine, No. 25 Heping North Road, Tianning District, Changzhou 213003, PR China; Jiangsu Key Laboratory for High Technology Research of TCM Formulae, National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine and Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, Nanjing University of Chinese Medicine, No.138, Xianlin Road, Qixia District, Nanjing 210023, PR China.
² Jiangsu Key Laboratory for High Technology Research of TCM Formulae, National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine and Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, Nanjing University of Chinese Medicine, No.138, Xianlin Road, Qixia District, Nanjing 210023, PR China.
³ Changzhou Key Laboratory of Human Use Experience Research & Transformation of Menghe Medical Sect, Changzhou Hospital of Chinese Medicinal Affiliated to Nanjing University of Chinese Medicine, No. 25 Heping North Road, Tianning District, Changzhou 213003, PR China.
⁴ Jiangsu Key Laboratory for High Technology Research of TCM Formulae, National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine and Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, Nanjing University of Chinese Medicine, No.138, Xianlin Road, Qixia District, Nanjing 210023, PR China. Electronic address: zhangyi@njucm.edu.cn.
⁵ Jiangsu Key Laboratory for High Technology Research of TCM Formulae, National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine and Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, Nanjing University of Chinese Medicine, No.138, Xianlin Road, Qixia District, Nanjing 210023, PR China. Electronic address: zhangli@njucm.edu.cn.
⁶ Changzhou Key Laboratory of Human Use Experience Research & Transformation of Menghe Medical Sect, Changzhou Hospital of Chinese Medicinal Affiliated to Nanjing University of Chinese Medicine, No. 25 Heping North Road, Tianning District, Changzhou 213003, PR China. Electronic address: liboluo2020@sina.com.
⁷ Changzhou Key Laboratory of Human Use Experience Research & Transformation of Menghe Medical Sect, Changzhou Hospital of Chinese Medicinal Affiliated to Nanjing University of Chinese Medicine, No. 25 Heping North Road, Tianning District, Changzhou 213003, PR China. Electronic address: czfy047@njucm.edu.cn.

PMID: 38677272
DOI: 10.1016/j.phymed.2024.155622

Abstract

Background: Cerebral ischemia-reperfusion injury (CIRI) is a destructive adverse reaction of ischemic stroke, leading to high disability and mortality rates. Salvia miltiorrhiza Bge. (Danshen, DS) processed with porcine cardiac blood (PCB-DS), a characteristic processed product, has promising anti-ischemic effects. However, the underlying mechanism of PCB-DS against CIRI remains unclear.

Purpose: Ferroptosis is demonstrated to be involved in CIRI. The aim of this study was to explore the molecular mechanism underlying PCB-DS inhibited GLRX5-mediated ferroptosis alleviating CIRI, which was different from DS.

Methods: Quality evaluation of PCB-DS and DS was conducted by UPLC. Pharmacological activities of PCB-DS and DS against CIRI were compared using neurobehavioral scores, infarct volume, proinflammatory factors, and pathological examinations. Proteomics was employed to explore the potential specific mechanism of PCB-DS against CIRI, which was different from DS. Based on the differential protein GLRX5, ferroptosis-related iron, GSH, MDA, SOD, ROS, liperfluo, and mitochondrial morphology were analyzed. Then, the proteins of GLRX5-mediated iron-starvation response and SLC7A11/GPX4 were analyzed. Finally, OGD/R-induced SH-SY5Y cells upon GLRX5 silencing were constructed to demonstrate that PCB-DS improved CIRI by GLRX5-mediated ferroptosis.

Results: PCB-DS better alleviated CIRI through decreasing neurological score, reducing the infarct volume, and suppressing the release of inflammatory cytokines than DS. Proteomics suggested that PCB-DS may ameliorate CIRI by inhibiting GLRX5-mediated ferroptosis, which was different from DS. PCB-DS reversed the abnormal mitochondrial morphology, iron, GSH, MDA, SOD, ROS, and liperfluo to inhibit ferroptosis in vitro and in vivo. PCB-DS directly activated GLRX5 suppressing the iron-starvation response and downregulated the SLC7A11/GPX4 signaling pathway to inhibit ferroptosis. Finally, silencing GLRX5 activated the iron-starvation response in SH-SY5Y cells and PCB-DS unimproved OGD/R injury upon GLRX5 silencing.

Conclusion: Different from DS, PCB-DS suppressed ferroptosis to alleviate CIRI through inhibiting GLRX5-mediated iron-starvation response. These findings give a comprehensive understanding of the molecular mechanism underlying the effect of PCB-DS against CIRI and provide evidence to assess the product in clinical studies.

Keywords: Cerebral ischemia-reperfusion injury; Ferroptosis; Iron-starvation response; Iron-sulfur cluster; Salvia miltiorrhiza Bge. proceed with Porcine cardiac blood.