摘要
[背景]铝激活信号转导和转录激活因子3(STAT3)致小胶质细胞活化核苷酸结合和寡聚化结构域样受体蛋白3(NLRP3)炎症小体产生炎症反应并造成神经毒性。[目的]探讨STAT3调控NLRP3炎症小体在麦芽酚铝(Al(mal)_(3))致小鼠小胶质细胞株(BV2)细胞炎症反应中的作用。[方法]选取BV2细胞株,利用Al(mal)_(3)染毒和STAT3拮抗剂C188-9干预,实验分为5组:对照组,Al(mal)_(3)低、中、高剂量组(40、80和160μmol·L^(-1)Al(mal)_(3)),C188-9干预组(10μmol·L^(-1)C188-9+160μmol·L^(-1)Al(m al)_(3))。采用CCK8检测细胞活力;采用Western blotting检测BV2细胞M1/M2型标志物CD68/CD206的表达,以及STAT3、p-STAT3、NLRP3、cleaved-casepase-1、衔接蛋白凋亡相关斑点样蛋白(ASC)表达量;采用ELISA检测促炎因子白细胞介素(IL)-1β、IL-18和抗炎因子IL-10的含量。[结果]细胞活力测定显示:随着染铝浓度的增加,各剂量组细胞活力逐渐降低。与对照组相比,Al(mal)_(3)高剂量组细胞活力下降18%(P<0.05);与Al(mal)_(3)高剂量组相比,C188-9干预组细胞活力升高14%(P<0.05)。与对照组相比,Al(mal)_(3)低、中、高剂量组CD68的表达分别升高19%、20%、21%(P<0.05),Al(mal)_(3)高剂量组CD206的表达降低25%(P<0.05)。与Al(mal)_(3)高剂量组相比,C188-9干预组CD68的表达水平降低9%(P<0.05),而CD206的表达水平升高22%(P<0.05)。与对照组相比,Al(mal)_(3)高剂量组p-STAT3蛋白表达量和p-STAT3/STAT3值分别增加129%和127%(P<0.05)。与Al(mal)_(3)高剂量组相比,C188-9干预组p-STAT3蛋白表达量和p-STAT3/STAT3值分别降低55%和54%(P<0.05)。NLRP3炎症小体测试结果显示,与对照组相比,Al(mal)_(3)高剂量组NLRP3蛋白表达量增加75%(P<0.05),Al(mal)_(3)中、高剂量组cleaved-casepase-1蛋白表达量分别增加28%、35%(P<0.05),Al(mal)_(3)低、中、高剂量组ASC表达量分别增加22%、25%、53%(P<0.05)。与Al(mal)_(3)高剂量组相比,C188-9干预组NLRP3、cleaved-casepase-1、ASC蛋白表达�
[Background]Aluminum activates signal transducer and activator of transcription 3(STAT3),causing microglial nucleotide-binding and oligomerization domain-like receptors protein 3(NLRP3)inflammasome activation and inflammatory responses and producing neurotoxicity.[Objective]To explore the role of STAT3 regulated NLRP3 inflammasomes in the inflammatory response of mouse microglia cell line(BV2)cells induced by maltol aluminum[Al(mal)_(3)].[Methods]BV2 cells were assigned to five groups:one control group,three Al(mal)_(3)exposure groups(low,medium,and high doses at 40,80,and 160μmol·L^(-1)Al(mal)_(3)respectively),and one C188-9(STAT3 antagonist)intervention group[10μmol·L^(-1)C188-9+160μmol·L^(-1)Al(mal)_(3)].Cell viability was detected by CCK8.The expression of M1/M2 type markers,i.e.CD68/CD206,STAT3,p-STAT3,NLRP3,cleaved-casepase-1,and apoptosis-associated speck-like protein(ASC)in BV2 cells were detected by Western blotting,and proinflammatory cytokines interleukin(IL)-1βand IL-18,and anti-inflammatory cytokine IL-10 were determined by ELISA.[Results]The results of cell viability assay showed that cell viability gradually decreased with the increase of Al(mal)_(3)dose.Compared with the control group,the cell viability of the Al(mal)_(3)high-dose group was decreased by 18%(P<0.05);compared with the Al(mal)_(3)high-dose group,the cell viability of the C188-9 intervention group was significantly elevated by 14%(P<0.05).Compared with the control group,the expression levels of CD68 in the Al(mal)_(3)low-,medium-,and high-dose groups were elevated by 19%,20%,and 21%,respectively(P<0.05);the expression level of CD206 in the Al(mal)_(3)high-dose group was decreased by 25%(P<0.05).Compared with the Al(mal)_(3)high-dose group,the expression level of CD68 in the C188-9 intervention group was reduced by 9%(P<0.05),whereas the expression level of CD206 was elevated by 22%(P<0.05).Compared with the control group,the p-STAT3 protein expression and the p-STAT3/STAT3 ratio in the Al(mal)_(3)high-dose group increased by 12
作者
王天枢
高丹
赵丹
宦佳萍
韩笑
宋静
王林平
张慧芳
牛侨
路小婷
WANG Tianshu;GAO Dan;ZHAO Dan;HUAN Jiaping;HAN Xiao;SONG Jing;WANG Linping;ZHANG Huifang;NIU Qiao;LU Xiaoting(School of Public Health,Shanxi Medical University,Taiyuan,Shanxi 030001,China)
出处
《环境与职业医学》
CSCD
北大核心
2023年第11期1250-1256,共7页
Journal of Environmental and Occupational Medicine
基金
山西省自然科学基金项目(202103021224226)
山西省回国留学人员科研资助项目(2021-084)。
关键词
麦芽酚铝
BV2细胞
信号转导和转录激活因子3
寡聚化结构域样受体蛋白3
maltol aluminum
BV2 cell
signal transducer and activator of transcription 3
nucleotide-binding and oligomerization domainlike receptors protein 3
