【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^-1C188-9+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（m al）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（m al）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（m al）₃高剂量组NLRP3蛋白表达量增加75%（P<0.05）,Al（m al）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蛋白表达量分别降低30%、19%、32%（P<0.05）。与对照组相比,IL-1β在Al（mal）₃中、高剂量组含量分别增加18%、21%（P<0.05）,IL-18在Al（mal）₃高剂量组的含量增加10%（P<0.05）。与Al（m al）₃高剂量组相比,C188-9干预组IL-18的含量降低23%（P<0.05）。抗炎因子IL-10的含量在各组差异无统计学意义（P>0.05）。[结论]铝能引起BV2小胶质细胞的炎症反应并且以促炎反应为主,其机制可能与STAT3调控NLRP3炎症小体分泌炎症因子有关。更多还原

【Abstract】 [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 160 μmol·L^-1 Al（mal）₃ respectively),and one C188-9（STAT3 antagonist） intervention group [10 μmol·L^-1 C188-9+160 μmol·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）-1β and 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）₃high-dose group,the cell viability of the C188-9 intervention group was significantly elevated by 14%（P <0.05）.Compared with the control gro up,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）₃high-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/STAT3ratio in the Al（mal）₃ high-dose group increased by 129% and 127%,respectively（P<0.05）.Compared with the Al（mal）₃ high-dose group,the p-STAT3 protein expression and the p-STAT3/STAT3 ratio in the C188-9 intervention group were decreased by 55% and 54%,respectively（P>0.05）.Compared with the control group,the expression level of NLRP3 protein increased by 75% in the Al（mal）₃ high-dose group（P<0.05）,the expression levels of cleaved-casepase-1 protein increased by 28% and 35% in the Al（mal）₃ medium-and high-dose groups（P<0.05）,and the expression levels of ASC increased by 22%,25%,and 53% in the Al（mal）₃ low-,medium-and high-dose groups（P <0.05）,respectively.Compared with the Al（mal）₃ high-dose group,the expression levels of NLRP3,cleaved-casepase-1,and ASC proteins in the C188-9 intervention group decreased by 30%,19%,and 32%,respectively（P<0.05）.Compared with the control group,the levels of IL-1β in the Al（mal）₃ medium-and high-dose groups increased by 18% and 21%,respectively（P<0.05）,and the level of IL-18 in the Al（mal）₃ high-dose group increased by 10%（P<0.05）.Compared with the Al（mal）₃ high-dose group,the IL-18 levels were reduced by 23%in the C188-9 intervention group（P<0.05）.The content of anti-inflammatory factor IL-10 did not differ significantly between groups（P>0.05）.[Conclusion] Aluminum can induce inflammatory responses in BV2 microglia and is predominantly pro-inflammato ry,and the mechanism may involve STAT3 regulation of NLRP3 inflammasome secretion of inflammatory factors.更多还原