摘要
目的:初步探讨大气细颗粒物(fine particulate matter,PM_(2.5))诱导大鼠肺泡巨噬细胞内NLRP3炎性小体活化及细胞凋亡的潜在分子机制。方法:采用中流量采样器收集PM_(2.5)颗粒物,经超声提取制成混悬液,作用于大鼠肺泡巨噬细胞(NR8383)后,MTT法检测细胞存活率,Western blot检测Nod样受体蛋白3(NLRP3)、组织蛋白酶B(Cathepsin-B)及凋亡相关蛋白[Bax、Bcl-2及半胱天冬酶-3(Caspase-3)]的表达水平,ELISA检测PM_(2.5)诱导的细胞上清液中半胱天冬酶-1(Caspase-1)、白介素(interleukin,IL)-18和IL-1β的含量。结果:随着PM_(2.5)浓度的增加和刺激时间的延长,NR8383细胞存活率降低(24 h:F=17.253,P=0.000;48 h:F=18.678,P=0.000;72 h:F=256.104,P=0.000);PM_(2.5)诱导NR8383细胞内NLRP3(F=437.166,P=0.000)和Cathepsin-B(F=42.062,P=0.000)蛋白的表达上调;PM_(2.5)上调细胞内促凋亡蛋白Bax(F=72.827,P=0.000)表达,下调抗凋亡蛋白Bcl-2(F=390.322,P=0.000)表达,下调Caspase-3(F=169.833,P=0.000)蛋白的表达;PM_(2.5)上调细胞培养液中Caspase-1(F=629.866,P=0.000)、IL-18(F=587.165,P=0.000)和IL-1β(F=68.472,P=0.000)的水平。结论:PM_(2.5)可通过溶酶体模式诱导肺泡巨噬细胞内NLRP3炎性小体活化,促进IL-18和IL-1β分泌,并引起肺泡巨噬细胞发生凋亡。
Objective:To investigate the potential molecular mechanism of fine particulate matter(PM_(2.5))on activation of NLRP3 inflammasome and apoptosis in rat alveolar macrophages.Methods:Rat alveolar macrophages were treated with PM2.5 suspension collected by medium flow atmospheric sampler.After action of the suspension on rat alveolar macrophages(NR8383),MTT assay was used to detect the cell survival rate.Western blot was performed to detect the expression of NLRP3,Cathepsin-B and apoptosis-related proteins(Bax,Bcl-2,Caspase-3).The level of Caspase-1,interleukin-18(IL-18)and interleukin-1β(IL-1β)in the cell supernatant induced by PM_(2.5)were measured by ELISA method.Results:With the increase of PM2.5 concentration and the extension of stimulation time,the survival rate of NR8383 decreased(24 h:F=17.253,P=0.000;48 h:F=18.678,P=0.000;72 h:F=256.104,P=0.000);PM_(2.5)accelerated the expression of NLRP3(F=437.166,P=0.000)and Cathepsin-B(F=42.062,P=0.000)in NR8383;PM_(2.5)up-regulated the expression of pro-apoptotic protein Bax(F=72.827,P=0.000),down-regulated the expression of anti-apoptotic protein Bcl-2(F=390.322,P=0.000),and down-regulated the expression of Caspase-3(F=169.833,P=0.000);PM_(2.5)increased the level of Caspase-1(F=629.866,P=0.000),IL-18(F=587.165,P=0.000)and IL-1β(F=68.472,P=0.000)in cell culture medium.Conclusion:PM_(2.5)can activate NLRP3 inflammasome in alveolar macrophages through the lysosomal model,promote the secretion of IL-18 and IL-1β,and cause apoptosis of alveolar macrophages.
作者
顾娜
谭珵
张桂贤
史鹏程
刘伟伟
赵秀梅
刘洪斌
田瑛泽
石钏
Gu Na;Tan Cheng;Zhang Guixian;Shi Pengcheng;Liu Weiwei;Zhao Xiumei;Liu Hongbin;Tian Yingze;Shi Chuan(Tianjin Institute of Medical and Pharmaceutical Sciences;College of Env ironme ntal Science and Engine ering,Nank ai University;Tianjin QuantoBio Biotech Co.,Ltd.)
出处
《重庆医科大学学报》
CAS
CSCD
北大核心
2021年第8期915-920,共6页
Journal of Chongqing Medical University
基金
天津市卫计委科技基金资助项目(编号:2015KY39)。
