Induction of ferroptosis in response to graphene quantum dots through mitochondrial oxidative stress in microglia

Abstract

Background Graphene quantum dots (GQDs) provide a bright prospect in the biomedical application because they contain low-toxic compounds and promise imaging of deep tissues and tiny vascular structures. However, the biosafety of this novel QDs has not been thoroughly evaluated, especially in the central nervous system (CNS). The microarray analysis provides a hint that nitrogen-doped GQDs (N-GQDs) exposure could cause ferroptosis in microglia, which is a novel form of cell death dependent on iron overload and lipid peroxidation. Results The cytosolic iron overload, glutathione (GSH) depletion, excessive reactive oxygen species (ROS) production and lipid peroxidation (LPO) were observed in microglial BV2 cells treated with N-GQDs, which indicated that N-GQDs could damage the iron metabolism and redox balance in microglia. The pre-treatments of a specific ferroptosis inhibitor Ferrostatin-1 (Fer-1) and an iron chelater Deferoxamine mesylate (DFO) not only inhibited cell death, but also alleviated iron overload, LPO and alternations in ferroptosis biomarkers in microglia, which were caused by N-GQDs. When assessing the potential mechanisms of N-GQDs causing ferroptosis in microglia, we found that the iron content, ROS generation and LPO level in mitochondria of BV2 cells all enhanced after N-GQDs exposure. When the antioxidant ability of mitochondria was increased by the pre-treatment of a mitochondria targeted ROS scavenger MitoTEMPO, the ferroptotic biological changes were effectively reversed in BV2 cells treated with N-GQDs, which indicated that the N-GQDs-induced ferroptosis in microglia could be attributed to the mitochondrial oxidative stress. Additionally, amino functionalized GQDs (A-GQDs) elicited milder redox imbalance in mitochondria and resulted in less ferroptotic effects than N-GQDs in microglia, which suggested a slight protection of amino group functionalization in GQDs causing ferroptosis. Conclusion N-GQDs exposure caused ferroptosis in microglia via inducing mitochondrial oxidative stress, and the ferroptotic effects induced by A-GQDs were milder than N-GQDs when the exposure method is same. This study will not only provide new insights in the GQDs-induced cell damage performed in multiple types of cell death, but also in the influence of chemical modification on the toxicity of GQDs.

Ferroptosis Enhanced Diabetic Renal Tubular Injury via HIF-1α/HO-1 Pathway in db/db Mice

“…However, Ferrostatin-1 can form a complex compounded with iron ( 45 ). Previous studies have found that ferroptosis inhibitor Ferrostatin-1 alleviated atherosclerosis lesion and inhibited the iron accumulation in HFD-fed ApoE -/- mice ( 46 ), protected against early brain injury and decreased the iron content in subarachnoid hemorrhage rats ( 47 ), and inhibited death of microglia and alleviated iron overloading induced by nitrogen-doped graphene quantum dots ( 48 ). In current research, Lillie staining showed that there was more iron accumulation in diabetic mouse renal tubules compared with non-diabetic mouse renal tubules, but iron overloading was relieved in renal tubules of diabetic mice after Ferrostatin-1 treatment.…”

Research advance on cell imaging and cytotoxicity of different types of quantum Dots

“…Thus, ROS induced by GQDs may increase the phosphorylation of p38-MAPK and promote the nuclear translocation of NF-κB, further lead to inflammation, apoptosis and autophagy (Qin, Zhou, et al, 2015). Recently, Wu et al found that nitrogen-doped GQDs (N-GQDs) can induce mitochondrial oxidative stress in microglia, which eventually lead to ferroptosis (Wu et al, 2020).…”

Direct and Indirect Genotoxicity of Graphene Family Nanomaterials on DNA—A Review

“…Organisms possess a well-developed inhibition of antioxidant defense, including ROSscavenging enzymes (e.g., superoxide dismutase (SOD), peroxidase, and catalase) and regulatory mechanisms to protect organisms from the negative effects of ROS [46,84]. The ROS generation benefitted from inhibition of fatty acid, carbohydrate, and amino acid metabolism [85]. ROS induced by GO seemed to be the main mechanism leading to human lung fibroblast (HLF) cells of genotoxicity [86].…”