Artemisinin Alleviates Cerebral Ischemia/Reperfusion-Induced Oxidative Damage via Regulating PHB2-Mediated Autophagy in the Human Neuroblastoma SH-SY5Y Cell Line

Abstract

Oxidative stress plays a key role in cerebral ischemia/reperfusion injury. Artemisinin (ART) has antioxidative stress activity in addition to its powerful antimalarial effects. In this article, we investigated the effect of ART on OGD/R-induced oxidative stress injury and its underlying mechanisms. We used oxygen-glucose deprivation/reoxygenation (OGD/R) to establish an in vitro model of cerebral ischemia/reperfusion (I/R) injury. CCK-8 and lactate dehydrogenase (LDH) release were used to assess cellular damage. Measurement of reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD), glutathione (GSH), and mitochondrial membrane potential (MMP) estimates oxidative stress-induced damage and protection from ART effect. OGD/R treatment aggravated oxidative stress damage, whereas ART reversed the effects of OGD/R. Autophagy is closely related to oxidative stress; in order to confirm whether the antioxidative stress effect of ART is related to PHB2-mediated autophagy, we examined the protein expression of prohibitin 2 (PHB2), TOMM20, p62, and the conversion of microtubule-associated protein light chain 3I (LC3I) to LC3II and found that the protein expression of PHB2, TOMM20, p62, and LC3II/LC3I was significantly correlated with OGD/R treatment. The colocalization of PHB2 and LC3, TOMM20, and LC3 was reduced after OGD/R treatment, and ART reversed this change. After silencing PHB2, the protective effect of ART against OGD/R-induced oxidative stress injury was reduced, the protein expressions of PHB2, TOMM20 and LC3II/LC3I and the colocalization of PHB2 and LC3, TOMM20, and LC3 were decreased. We used chloroquine to block the lysosomal pathway and found that ART increased the conversion of LC3I to LC3II, silencing PHB2 which inhibited the conversion of LC3I to LC3II, and impaired mitophagy. Our findings showed that ART attenuated OGD/R-induced oxidative stress damage through PHB2-mediated mitophagy. To the current knowledge, our study is the first to demonstrate that ART attenuates OGD/R-induced oxidative stress injury through PHB2-mediated autophagy in the human neuroblastoma SH-SY5Y cell line, which provided new insights into the treatment of OGD/R injury.

A study on the mechanism of Beclin-1 m6A modification mediated by catalpol in protection against neuronal injury and autophagy following cerebral ischemia

“…For instance, Yang et al pointed out that Arctium lappa L. roots protected against cerebral ischemia by repressing apoptosis and AMPK/mTOR-mediated autophagy (Yang et al 2021a , b ). Jiang and his team specified that Artemisinin could attenuate OGD/R-induced oxidative stress damage through PHB2-mediated mitophagy (Jiang et al 2022 ). The active compounds in drugs can regulate the autophagy of various cells, including tumor cells, neurons, myocardial cells, and endothelial cells (Huang et al 2015 ).…”

PHB2 Alleviates Neurotoxicity of Prion Peptide PrP106–126 via PINK1/Parkin-Dependent Mitophagy

“…Lai and co-workers reported that Rutin, a natural botanical ingredient, attenuated oxidative damage, through PHB2-mediated mitophagy, in MPP + -treated SH-SY5Y cells [65]. Artemisinin, known for its powerful antioxidative stress effect, alleviated oxidative damage caused by cerebral ischemia/reperfusion by regulating PHB2-mediated autophagy in the human neuroblastoma SH-SY5Y cell line [66].…”

“…While further research is needed to confirm and explore the full role of PHB2 in prion diseases, our studies provide hope for targeting PHB2 as a therapeutic approach for neurodegenerative diseases associated with prions. [66].…”

Progress of medicinal plants and their active metabolites in ischemia-reperfusion injury of stroke: a novel therapeutic strategy based on regulation of crosstalk between mitophagy and ferroptosis

“…Recent research indicates that ART not only has a potent antimalarial effect ( Chaturvedi et al, 2010 ), but also exhibits antioxidant stress activity ( Ahmed-Laloui et al, 2022 ). As reported by Jiang MH et al ( Jiang et al, 2022 ), they constructed an in vitro model of cerebral ischemia-reperfusion injury by using OGD/R. Subsequently, they assessed cell damage by using CCK-8 and LDH release, and evaluated the oxidative stress-induced damage and ART’s protective effects by using measurements of ROS, MDA, SOD, GSH, and MMP.…”