摘要
目的:探讨木香烃内酯(Cos)对脂多糖(LPS)诱导的小鼠原代巨噬细胞炎性小体激活和炎症反应的影响。方法:提取并培养小鼠腹腔原代巨噬细胞,用不同浓度Cos预处理巨噬细胞1 h后,1 mg/L LPS处理,分为空白对照组、模型组(单用LPS)、不同浓度(5、10和20μmol/L) Cos+LPS组;ELISA法检测培养液上清中TNF-α/IL-6的含量,q PCR法检测TNF-α/IL-6 mRNA表达;为明确Cos对LPS诱发的炎症反应的保护机制,增加NLRP3抑制剂MCC950组(10μmol/L),LPS刺激4h、6 h,Western blot检测NLRP3、ERK及IκB的激活,细胞免疫荧光实验观察NLRP3的蛋白水平。结果:与LPS组比较,LPS+Cos组TNF-α/IL-6的mRNA表达及细胞培养液上清TNF-α/IL-6分泌均显著降低(P <0. 05);Western blot和细胞免疫荧光实验结果表明,与LPS组比较,LPS+Cos组的NLRP3蛋白水平显著降低,其抑制效果与NLRP3抑制剂MCC950相当(P <0. 05);Western blot结果表明,与LPS组比较,LPS+MCC950组和LPS+Cos组的ERK及IκB的激活均显著降低(P <0. 01)。结论:Cos能有效抑制LPS诱导的巨噬细胞炎性小体和NF-κB的激活而缓解炎症反应。
Objective:To investigate the effect of Costunolide on lipopolysaccharide(LPS)-induced activation of inflammatory corpuscles and inflammatory response in murine primary macrophages.Methods:Mouse peritoneal macrophages were extracted and cultured,and macrophages were divided into blank control group,model group(LPS),LPS+Costunolide group.After pretreatment with1 h,LPS was used to stimulate 6 h or 24 h,and total RNA or protein and culture medium were extracted.ELISA was used to detect the content of TNF-α/IL-6 in the culture medium,and q PCR was used to detect the expression of TNF-α/IL-6 mRNA.Western blot was used to detect the activation of NLRP3,ERK and IκB,and the level of NLRP3 protein was observed by cellular immunofluorescence assay.Results:Compared with group LPS,the mRNA expression and secretion of TNF-α/IL-6 significantly decreased in LPS+Costunolide group(P<0.001 or P<0.05).Experimental results of Western blot and immunofluorescence showed that,compared with LPS group,NLRP3 protein level in LPS+Costunolide group was significantly decreased(P<0.01).The results of Western blot showed that compared with LPS group,ERK and IκB activation in LPS+Costunolide group decreased significantly(P<0.01).Conclusion:Costunolide can effectively inhibit LPS induced macrophage inflammatory corpuscle and activation of NF-κB and alleviate inflammatory responses.
作者
吴晓燕
厉星
WU Xiao-Yan;LI Xing(Department of Pharmacy,Sir Run Run Shaw Hospital,School of Medicine,Zhejiang University,Hangzhou 310018,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2019年第12期1433-1437,共5页
Chinese Journal of Immunology
