Ginsenoside compound K protects against cerebral ischemia/reperfusion injury via Mul1/Mfn2-mediated mitochondrial dynamics and bioenergy

Qingxia Huang; Jing Li; Jinjin Chen; Zepeng Zhang; Peng Xu; Hongyu Qi; Zhaoqiang Chen; Jiaqi Liu; Jing Lu; Mengqi Shi; Yibin Zhang; Ying Ma; Daqing Zhao; Xiangyan Li

doi:10.1016/j.jgr.2022.10.004

Ginsenoside compound K protects against cerebral ischemia/reperfusion injury via Mul1/Mfn2-mediated mitochondrial dynamics and bioenergy

J Ginseng Res. 2023 May;47(3):408-419. doi: 10.1016/j.jgr.2022.10.004. Epub 2022 Oct 20.

Authors

Qingxia Huang^{1

2}, Jing Li², Jinjin Chen², Zepeng Zhang^{1

2}, Peng Xu³, Hongyu Qi², Zhaoqiang Chen², Jiaqi Liu², Jing Lu^{1

3}, Mengqi Shi⁴, Yibin Zhang³, Ying Ma³, Daqing Zhao², Xiangyan Li²

Affiliations

¹ Research Center of Traditional Chinese Medicine, College of Traditional Chinese Medicine, Changchun University of Chinese Medicine, Changchun, Jilin, China.
² Northeast Asia Research Institute of Traditional Chinese Medicine, Key Laboratory of Active Substances and Biological Mechanisms of Ginseng Efficacy, Ministry of Education, Jilin Provincial Key Laboratory of Bio-Macromolecules of Chinese Medicine, Changchun University of Chinese Medicine, Changchun, Jilin, China.
³ Department of Encephalopathy, College of Traditional Chinese Medicine, Changchun University of Chinese Medicine, Changchun, Jilin, China.
⁴ Department of Graduate Administration, College of Traditional Chinese Medicine, Changchun University of Chinese Medicine, Changchun, Jilin, China.

Abstract

Background: Ginsenoside compound K (CK), the main active metabolite in Panax ginseng, has shown good safety and bioavailability in clinical trials and exerts neuroprotective effects in cerebral ischemic stroke. However, its potential role in the prevention of cerebral ischemia/reperfusion (I/R) injury remains unclear. Our study aimed to investigate the molecular mechanism of ginsenoside CK against cerebral I/R injury.

Methods: We used a combination of in vitro and in vivo models, including oxygen and glucose deprivation/reperfusion induced PC12 cell model and middle cerebral artery occlusion/reperfusion induced rat model, to mimic I/R injury. Intracellular oxygen consumption and extracellular acidification rate were analyzed by Seahorse multifunctional energy metabolism system; ATP production was detected by luciferase method. The number and size of mitochondria were analyzed by transmission electron microscopy and MitoTracker probe combined with confocal laser microscopy. The potential mechanisms of ginsenoside CK on mitochondrial dynamics and bioenergy were evaluated by RNA interference, pharmacological antagonism combined with co-immunoprecipitation analysis and phenotypic analysis.

Results: Ginsenoside CK pretreatment could attenuate mitochondrial translocation of DRP1, mitophagy, mitochondrial apoptosis, and neuronal bioenergy imbalance against cerebral I/R injury in both in vitro and in vivo models. Our data also confirmed that ginsenoside CK administration could reduce the binding affinity of Mul1 and Mfn2 to inhibit the ubiquitination and degradation of Mfn2, thereby elevating the protein level of Mfn2 in cerebral I/R injury.

Conclusion: These data provide evidence that ginsenoside CK may be a promising therapeutic agent against cerebral I/R injury via Mul1/Mfn2-mediated mitochondrial dynamics and bioenergy.

Keywords: Bioenergy; Cerebral ischemia/reperfusion injury; Ginsenoside compound K; Mitochondrial dynamics; Mul1/Mfn2 ubiquitination.