抑制NLRP3炎症小体活化对二氧化硅粉尘致巨噬细胞炎性反应的影响

Inhibition of NLRP3 inflammasome activation on the inflammatory response of macrophage induced by silica dust

目的探讨抑制核苷酸结合寡聚化结构域受体家族含pyrin蛋白结构域蛋白3(NLRP3)炎症小体活化对二氧化硅(SiO2)粉尘致巨噬细胞炎性反应的影响。方法用大鼠肺泡巨噬细胞(NR8383)建立细胞模型,分为空白对照组、染尘组(50 mg/L矽尘混悬液)、NLRP3抑制剂组(50 mg/L矽尘混悬液和20 μmol/L NLRP3抑制剂)、木犀草素组(50 mg/L矽尘混悬液和20 μmol/L木犀草素)。培养12、24、48 h收集样本。用酶联免疫吸附实验(ELISA)检测白细胞介素(IL)、转化生长因子-β(TGF-β1)等细胞炎性因子分泌情况,用Western Blot法检测NLRP3和Caspase-1蛋白水平。结果与空白对照组比较,染尘组、NLRP3抑制剂组、木犀草素组NR8383细胞经染尘12、24、48 h后,细胞存活率均下降,IL-1β、IL-18、TGF-β1、NLRP3和Caspase-1表达水平均明显升高,差异均有统计学意义(P<0.05)。与染尘组比较,NLRP3抑制剂组、木犀草素组NR8383细胞经染尘12、24、48 h后,细胞存活率提高,IL-1β、IL-18、TGF-β1、NLRP3和Caspase-1水平均明显降低,差异均有统计学意义(P<0.05)。与NLRP3抑制剂组比较,木犀草素组NR8383细胞的IL-1β水平在12、24、48 h时均明显降低,IL-18、TGF-β1水平在48 h时明显降低,NLRP3和Caspase-1水平在24 h时明显降低,差异均有统计学意义(P<0.05)。结论抑制NLRP3炎症小体活化和木犀草素均可以降低SiO2粉尘引起的巨噬细胞炎性反应。

Objective To investigate the effect of inhibiting the activation of NLRP3 inflammatory bodies on the inflammatory response of macrophages induced by silica(SiO2)dust.Methods Rat alveolar macrophages(NR8383)cells were used to establish the cell model,which was divided into four groups:blank control group,dust exposure group(50 mg/L silica dust suspension),NLRP3 inhibitor group(50 mg/L silica dust suspension and 20μmol/L NLRP3 inhibitor),luteolin group(50 mg/L silica dust suspension and 20μmol/L luteolin).Samples were collected at 12,24 and 48 hours after culture.The secretion of inflammatory factors IL and TGF-β1 were detected by ELISA.The levels of NLRP3 and Caspase-1 were detected by Western Blot.Results Compared with the blank control group,the survival rates of NR8383 cells in the dust exposure group,NLRP3 inhibitor group and luteolin group were all decreased after 12,24 and 48 hours of dust exposure,and the expression levels of IL-1β,IL-18,TGF-β1,NLRP3 and Caspase-1 were significantly increased(P<0.05).Compared with the dust exposure group,NR8383 cells in NLRP3 inhibitor group and luteolin group had higher cell survival rates and lower levels of IL-1β,IL-18,TGF-β1,NLRP3 and Caspase-1 after 12,24 and 48 hours of dust exposure(P<0.05).Compared with the NLRP3 inhibitor group,the IL-1βlevel of NR8383 cells in luteolin group decreased significantly at 12,24 and 48 h,the IL-18 and TGF-β1 levels decreased significantly at 48 h,the NLRP3 and Caspase-1 levels decreased significantly at 24 h(P<0.05).Conclusion Inhibition of NLRP3 inflammaasome activation can reduce the inflammatory response of macrophages induced by SiO2 dust.