MoS_(2)纳米酶通过调控线粒体动力及自噬减轻炎性内皮细胞损伤

MoS_(2) nanozyme attenuated inflammation-related endothelial cell injury by regulating mitochondrial dynamics and mitophagy

探讨新型MoS_(2)纳米酶通过调控线粒体动力,以减轻炎性血管内皮细胞损伤的保护机制。利用水热法制备出了花状MoS_(2)纳米片,结合电子自旋共振(electron spin resonance,ESR)光谱检测技术,表明花状MoS_(2)纳米片对羟基自由基(·OH)和单线态氧(^(1)O_(2))都具有很强的清除能力,呈剂量依赖效应。通过体外脂多糖(lipopolysaccharide,LPS)诱导的血管内皮细胞炎性氧化应激损伤模型,用花状MoS_(2)纳米片预处理,本研究分别用MTT及Annexin V-FITC/PI双染法检测内皮细胞毒性及凋亡;用MitoTracker荧光探针观察内皮细胞线粒体分裂及融合形态;用活性氧(reactive oxygen species,ROS)探针DCFH-DA及超氧阴离子(O2-)探针DHE检测细胞氧化应激水平;用质粒GFP-LC3转染及荧光共定位技术观察并分析细胞自噬及线粒体自噬形成。结果表明,MoS_(2)纳米酶可以显著减少炎性内皮细胞的细胞毒性及细胞凋亡,减轻炎性内皮细胞线粒体的分裂,并维持融合状态下的线粒体动力;还可以缓解LPS介导的内皮细胞线粒体自噬,进而保护内皮细胞免受炎性氧化应激性损伤。以上结果确立了新型MoS_(2)纳米酶可以通过调控内皮细胞线粒体动力及线粒体自噬,实现对炎性内皮细胞损伤的保护,有望拓展MoS_(2)纳米酶用于防治慢性炎症性血管内皮损伤相关疾病。

To explore the protective mechanisms of a novel molybdenum disulfide(MoS_(2))nanozyme in alleviating inflammation-related endothelial cell injury by regulating mitochondrial dynamic,flower like-MoS_(2)nanosheets were prepared by hydrothermal method,and its antioxidant enzyme-mimic activities were assessed via electron spin resonance(ESR)spectroscopy.It was shown that MoS_(2)nanosheets had strong scavenging ability for hydroxyl radical(·OH)and singlet reactive oxygen species(^(1)O_(2))in a dose-dependent manner.Using an in vitro lipopolysaccharide(LPS)-induced vascular endothelial cell injury model,the protective roles of MoS_(2)nanozyme on cytotoxicity and apoptosis of endothelial cells were examined by MTT and Annexin V-FITC/PI assay,respectively.Mitochondrial fission/fusion of endothelial cell were observed by Mito-Tracker green probe.Reactive oxygen species(ROS)probe DCFH-DA and superoxide anion probe DHE were used to detect the level of oxidative stress in vitro.Plasmid GFP-LC3 transfection using colocalization analysis was applied to assess the autophagy of endothelial cells.The results showed that MoS_(2)nanozyme could significantly reduce the cytotoxicity and apoptosis of endothelial cells stimulated by LPS,and prevent the impairment mitochondrial dynamics of endothelial cells,thus maintaining mitochondrial dynamics.In addition,MoS_(2)nanozyme was also shown to alleviate LPS-mediated endothelial mitochondrial autophagy,thus protecting endothelial cells from inflammatory stress.These results established that MoS_(2)nanozyme protected endothelial cells injury from inflammatory stress by regulating mitochondrial dynamics and mitochondrial autophagy of endothelial cells,which is expected to expand the use of MoS_(2)nanozyme in the prevention and treatment of inflammation-related vascular endothelial diseases.