中国药理学与毒理学杂志2024，Vol.38Issue(5)：375-383,9.DOI:10.3867/j.issn.1000-3002.2024.05.007
SARS-CoV-2刺突糖蛋白对神经细胞的损伤效应及机制
Damage effect and mechanism of SARS-CoV-2 spike protein on nerve cells
摘要
Abstract
OBJECTIVE To investigate the damage effect and potential toxic mechanism of SARS-CoV-2 spike protein(S protein)on human neuroblastomacells(SH-SY5Y).METHODS SH-SY5Y were treated with S protein at concentrations of 25,50,75,and 100 mg·L-1 for 24 h.Cell viability of SH-SY5Y was detected using the CCK-8 assay.The cytotoxic lactate dehydrogenase(LDH)detection kit was used to measure the release rate of LDH,and the 5-ethynyl-2′-deoxyuridine(EdU)-488 cell prolifera-tion kit was used to assess cell proliferation.The ATP detection kit was used to measure intracellular ATP content.The JC-1 fluorescent probe method was employed to detect the mitochondrial membrane potential(MMP)of cells.Seahorse XF was used to measure mitochondrial respiratory and glycolytic capacity.RESULTS Compared with the cell control group,cell viability was significantly reduced in S protein 25,50,75 and 100 mg·L-1 groups(P<0.01),and the half-inhibition concentration(IC50)was 65.05 mg·L-1.The LDH release rate wassignificantly increased(P<0.01)and the proportion of EdU positive cellswas significantly reduced(P<0.01)in S protein 25,50,75 and 100 mg·L-1 groups.S protein signifi-cantly reduced intracellular ATP content(P<0.01)at the concentrations of 75 and 100 mg·L-1,while significantly reduced intracellular MMP(P<0.05,P<0.01)at the concentrations of 50 and 75 mg·L-1.S protein 50 mg·L-1 increased the maximum value of basal glycolysis levels and glycolytic capacity(P<0.05,P<0.01),and S protein 25 and 50 mg·L-1 increased the maximum value of respiration capacity(P<0.05,P<0.01).SH-SY5Y cell viability was positively correlated with the intracellular ATP content and the MMP level(r2=0.9209,P=0.001;r2=0.6170,P=0.0025),and negatively correlated with the maximum level of basal glycolysis and glycolytic capacity(r2=0.5194,P=0.0285;r2=0.6664,P=0.0073),and nega-tively correlated with ATP production capacity(r2=0.8204,P=0.0008).CONCLUSIONS protein decreases the viability of SH-SY5Y cells and inhibited cell proliferation.The mechanism may be closely related to the disorder of energy metabolism.关键词
严重急性呼吸综合征冠状病毒2(SARS-CoV-2)/刺突糖蛋白/人神经母细胞瘤细胞/神经损伤/糖酵解/能量代谢Key words
severe acute respiratory syndrome coronavirus 2/spike protein/human neuroblastoma cells/injury of nerve/cell viability/glycolysis/energy metabolism分类
医药卫生引用本文复制引用
王娇,李佳佳,肖文一,韦冬晖,蒋宁,周文霞..SARS-CoV-2刺突糖蛋白对神经细胞的损伤效应及机制[J].中国药理学与毒理学杂志,2024,38(5):375-383,9.基金项目
国家重点研发计划课题(2022YFC3500304) （2022YFC3500304）
省部共建组分中药国家重点实验室开放课题(CBCM2023101) National Key R&D Program of China(2022YFC3500304) （CBCM2023101）
and State Key Laboratory of Component-based Chinese Medicine(CBCM2023101) （CBCM2023101）
