摘要
目的探讨氧化锌纳米颗粒(Zinc oxide nanoparticles,ZnO NPs)对小鼠小胶质细胞HMGB1/TLR4/RAGE信号通路的影响,以及HMGB1/TLR4/RAGE信号通路在ZnO NPs导致的小鼠小胶质细胞炎症反应和细胞焦亡中的可能作用。方法采用ZnO NPs处理BV2小鼠小胶质细胞24 h,MTT法检测ZnO NPs对小鼠小胶质细胞增殖活性影响;LDH实验检测细胞膜完整性;ELISA法检测细胞培养基中IL-1β分泌水平;Western blot法检测HMGB1、TLR4、RAGE、Caspase-1和GSDMD蛋白表达水平。结果ZnO NPs染毒24 h后,小鼠小胶质细胞出现明显的生长抑制(P<0.05),细胞培养液上清LDH活性升高,细胞内HMGB1、TLR4和RAGE蛋白表达增加(P<0.05),而用丙酮酸乙酯(Ethyl Pyruate,EP)处理后,上述蛋白含量均下降,差异有统计学意义(P<0.05);细胞内Caspase-1、GSDMD和GSDMD-N水平升高(P<0.05),细胞上清液中IL-1β分泌水平增加(P<0.05),经EP处理后,这些损伤均有一定程度改善。结论ZnO NPs可抑制小鼠小胶质细胞活力,造成细胞膜损伤,且可能通过HMGB1/TLR4/RAGE信号通路的激活介导炎症反应和细胞焦亡的发生。
Objective To explore the effects of ZnO NPs on the HMGB1/TLR4/RAGE signaling pathway in BV2 mouse microglial cells,to clarify the role of HMGB1/TLR4/RAGE signaling pathway in the inflammation and pyroptosis induced by ZnO NPs in BV2 cells.Methods BV2 mouse microglial cells were treated with ZnO NPs for 24 h.The cell viability was detected by MTT assay.The cell membrane integrity of microglia was detected by the lactate dehydrogenase(LDH)test.The secretion of IL-1βin cell supernatant was detected by ELISA.The expressions of HMGB1,TLR4,RAGE,Caspase-1,and GSDMD were determined by Western blot.Results After exposure to zinc oxide nanoparticles for 24 h,the cell viability was decreased(P<0.05),and the LDH level in the cell medium was enhanced(P<0.05).The protein levels of HMGB1,TLR4,and RAGE in cells increased and decreased when treated with ethyl pyruate(P<0.05).The protein contents of Caspase-1,GSDMD,GSDMD-N were elevated(P<0.05),and the secretion of inflammatory factor IL-1βwas enhanced(P<0.05).After treatment with EP,these injuries were rescued.Conclusion ZnO NPs might decrease cell viability,damage cell membrane,and promote the inflammation and pyroptosis of BV2 microglia cells,which might be mediated by the activation of the HMGB1/TLR4/RAGE signaling pathway.
作者
孙腾腾
张小强
崔丹丹
黄文迪
张秀文
SUN Teng-teng;ZHANG Xiao-qiang;CUI Dan-dan;HUANG Wen-di;ZHANG Xiu-wen(School of Public Health,Southeast University,Nanjing Jiangsu 210009,China;Key Laboratory of Environmental Medicine,Ministry of Education,Nanjing Jiangsu 210009,China)
出处
《毒理学杂志》
CAS
CSCD
2022年第4期304-309,315,共7页
Journal of Toxicology
基金
江苏省普通高校研究生科研创新计划资助项目(SJZZ16)。
