摘要
目的探讨沙眼衣原体质粒蛋白Pgp3诱导HeLa细胞产生IL-8的机制。方法选择不同质量浓度的Pgp3蛋白刺激HeLa细胞并在不同时间点收集细胞,Real-time PCR检测Pgp3刺激HeLa细胞后IL-8转录水平,Western blot和Real-time PCR分别检测NOD1的表达水平及下游蛋白RIP2的转录水平;Western blot检测ERK1/2和p38磷酸化水平;HeLa细胞用NOD1抑制剂ML130、RIP2抑制剂GSK583、ERK抑制剂PD98059和p38抑制剂LY2228820分别预处理后,再用Pgp3蛋白刺激细胞,ELISA和Real-time PCR检测IL-8的表达水平。结果 0~24μg/ml的Pgp3蛋白刺激HeLa细胞后,IL-8的含量呈时间和质量浓度依赖性,Pgp3蛋白刺激HeLa细胞能调控NOD1及RIP2的表达,同时ERK1/2和p38磷酸化水平显著升高;NOD1和RIP2抑制剂并不影响Pgp3蛋白诱导的ERK和p38磷酸化水平;用4种抑制剂预处理HeLa细胞后,Pgp3刺激HeLa细胞,IL-8表达量均出现下降。结论 Pgp3经NOD1/RIP2以及活化ERK1/2和p38信号通路诱导HeLa细胞调控IL-8的产生。
To investigate the mechanism of Chlamydia trachomatis plasmid-encoded Pgp3 inducing the production of IL-8 in He La cells, several concentrations of Pgp3 were selected to stimulate He La cells in different time course, and then the IL-8 expression level, NOD1 and RIP2 transcriptional levels were detected via real-time PCR and Western blotting. Besides, Western blot was used to detect ERK1/2 and p38 phosphorylating on various time course. Data showed that Pgp3 elicited IL-8 production in He La cells in a way of time and concentration dependence, and the optimal concentration and time was 24 μg/ml and 24 h, respectively; Pgp3 could modulate the expression of NOD1/RIP2 and phosphorylate ERK1/2 and p38, which would not affect by pretreatment of NOD1 inhibitor ML130, RIP2 inhibitor GSK583. And NOD1 inhibitor ML130, RIP2 inhibitor GSK583, ERK inhibitor PD98059 or p38 inhibitor LY2228820 could reverse the up-regulation of IL-8 induced by Pgp3 stimulation. Taken together, Pgp3 protein induces IL-8 production in He La cells through NOD1/RIP2 in ERK or p38-independent pathway.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2017年第9期760-764,共5页
Immunological Journal
基金
湖南省研究生科研创新项目(CX2016B475)
南华大学博士启动基金(2016XQD22)
国家自然科学基金(81072417)
