Caspase-4非经典炎症小体对问号钩体诱导THP-1细胞炎症因子分泌水平的影响  

Effects of Caspase-4 non-canonical inflammasome on the secretion of inflammatory cytokines in THP-1 cells induced by Leptospira interrogans

作　　者：汪小娟 刘英[2] 王铭 肖子宇 杨幸贵 洪峰 李世军 WANG Xiao-juan;LIU Ying;WANG Ming;XIAO Zi-yu;YANG Xing-gui;HONG Feng;LI Shi-jun(School of Public Health,Guizhou Medical University,Guiyang,Guizhou 550025,China;不详)

机构地区：[1]贵州医科大学公共卫生学院,贵州贵阳550025 [2]贵州省疾病预防控制中心细菌性疾病检验科

出　　处：《中国微生态学杂志》2022年第4期394-399,共6页Chinese Journal of Microecology

基　　金：国家自然科学基金(81760366,81960622);贵州省科技厅资助培养项目[黔科合(2016)4021];贵州省科技厅资助培养项目(黔科合平台人才[2018]5606号)。

摘　　要：目的了解Caspase-4非经典炎症小体在问号钩体诱导THP-1细胞炎性细胞因子分泌过程中的作用。方法采用问号钩体感染THP-1细胞(预先经佛波酯刺激分化为巨噬细胞)建立细胞模型,用实时荧光PCR扩增检测caspase-4、IL-18、IL-1β和IL-1αmRNA水平,Western blotting检测Caspase-4蛋白表达,ELISA定量检测细胞上清中IL-18、IL-1β和IL-1α分泌情况。结果实时荧光PCR和Western blotting显示,与未感染细胞比较,THP-1细胞Caspase-4 mRNA及蛋白表达水平均升高(t=46.03、29.36,均P<0.05),Caspase-4 siRNA转染后,Caspase-4 mRNA及蛋白表达水平显著下降(t=32.48、30.77,均P<0.01);钩体感染后,IL-18、IL-1β和IL-1αmRNA水平显著升高(t=25.70、26.13、19.94,均P<0.05),在Caspase-4特异性阻断后显著下降(t=11.55、44.68、15.68,均P<0.05);ELISA检测显示,钩体感染后,IL-18、IL-1β和IL-1α分泌均显著升高(t=50.24、34.17、25.18,均P<0.05),且能被Caspase-4特异性阻断剂有效抑制(t=42.00、17.07、5.03,均P<0.05)。结论Caspase-4非经典炎症小体参与介导问号钩体诱导人单核-巨噬细胞炎性细胞因子IL-18、IL-1β和IL-1α的分泌。Objective To observe the effects of Caspase-4 non-canonical inflammasome on the secretion of inflammatory cytokines of THP-1 cells induced by Leptospira interrogans(L.interrogans).Methods THP-1 cells were infected with L.interrogans to establish cell model.Real-time PCR were used to detect the mRNA levels of Caspase-4,IL-18,IL-1βand IL-1α.The expression of Caspase-4 was detected by using Western blotting.ELISA was used to detect the secretion of IL-18,IL-1βand IL-1αin the supernatants.Results The Caspase-4 mRNA and protein expression levels of THP-1 cells were up-regulated after infected by L.interrogans(t=46.03,t=29.36,all P<0.05),and decreased after Caspase-4 siRNA transfection(t=32.48,t=30.77,all P<0.01).Real-time PCR results indicated that the mRNA levels of IL-1β,IL-1αand IL-18 of THP-1 cells were significantly up-regulated after infected by L.interrogans(t=25.70,t=26.13,t=19.94,all P<0.05),and decreased significantly after inhibited by Caspase-4 inhibitor(t=11.55,t=44.68,t=15.68,all P<0.05).ELISA detection demonstrated that the secretion levels of IL-1β,IL-1αand IL-18 significantly rose after infected by L.interrogans(t=50.24,t=34.17,t=25.18,all P<0.05),and could be effectively inhibited by Caspase-4 inhibitor(t=42.00,t=17.07,t=5.03,all P<0.05).Conclusion Caspase-4 non-canonical inflammasome participates in the mediation of IL-1β,IL-1αand IL-18 secretion of THP-1 cells induced by L.interrogans.

关 键 词：问号钩体 细胞因子 单核-巨噬细胞 Caspase-4非经典炎症小体 

分 类 号：R377.5[医药卫生—病原生物学]