沙眼衣原体T3SS效应蛋白CT622通过激活ERK/p38 MAPK信号通路抑制Hela299细胞凋亡  

Chlamydia trachomatis T3SS effector protein CT622 inhibits Hela299 cell apoptosis by activating ERK/p38 MAPK signaling pathway

作　　者：胡艺宝 彭彬峰 曹倩 雷文波 刘双全 HU Yibao;PENG Binfeng;CAO Qian;LEI Wenbo;LIU Shuangquan(Department of Clinical Laboratory Medicine,Institution of Microbiology and Infectious Diseases,the First Affiliated Hospital,Hengyang Medical School,University of South China,Hengyang 421001,Hunan,China;Center of Laboratory Medicine and Pathology,the First Affiliated Hospital of Hunan University of Traditional Chinese Medicine,Changsha 410000,Hunan,China)

机构地区：[1]南华大学衡阳医学院附属第一医院检验医学中心,微生物与感染性疾病研究所,湖南衡阳421001 [2]湖南中医药大学第一附属医院医学检验与病理中心,湖南长沙410000

出　　处：《中南医学科学杂志》2024年第3期324-328,共5页Medical Science Journal of Central South China

基　　金：湖南省自然科学基金(2022JJ30532,2023JJ30540);湖南省高发性传播疾病精准诊治临床医学研究中心项目(2023SK4051);湖南省研究生科研创新项目基金(CX20230999);南华大学临床“4310”项目(20224310NHYCG02)。

摘　　要：目的探究沙眼衣原体Ⅲ型分泌系统(T3SS)效应蛋白CT622对Hela299细胞凋亡的影响机制。方法设置不同质量浓度CT622蛋白刺激Hela299细胞,筛选最佳刺激质量浓度。将细胞实验分为PBS组(空白对照)、肿瘤坏死因子(TNF)-α组(阳性对照)、CT622组、CT622+TNF-α组、CT622+PD98059(ERK1/2抑制剂)组、CT622+SB203580(p38抑制剂)组、CT622+TNF-α+PD98059组、CT622+TNF-α+SB203580组。Hoechst33258染色和流式细胞术检测Hela299细胞的凋亡情况。Western blotting检测凋亡相关蛋白裂解的Caspase-3(cleaved-Casepase-3)、B细胞淋巴瘤(Bcl)-2、Bcl-2相关X蛋白(Bax)和凋亡信号通路中信号分子ERK/p38 MAPK蛋白的表达。结果5 mg/L CT622蛋白为Hela299细胞最佳刺激质量浓度。CT622蛋白显著抑制Hela299细胞的凋亡,并上调ERK1/2、p38 MAPK和Bcl-2蛋白表达,下调Bax和Caspase-3蛋白表达。分别利用ERK1/2和p38 MAPK抑制剂与CT622联合处理可以明显增加Hela299细胞凋亡程度。结论T3SS效应蛋白CT622通过激活ERK/p38 MAPK信号通路抑制Hela299细胞凋亡。Aim To investigate the mechanism of Chlamydia trachomatis type 3 secretion system(T3SS)effector protein CT622 to affect Hela299 cell apoptosis.Methods Different mass concentrations of CT622 protein were used to stimulate Hela299 cells and to screen for the optimal stimulation mass concentration.Cell experiments were divided into PBS group(blank control),tumor necrosis factor(TNF)-αgroup(positive control),CT622 group,CT622+PD98059(ERK1/2 inhibitor)group,CT622+SB203580(ERK1/2 inhibitor)group,CT622+TNF-α+PD98059 group,and CT622+TNF-α+SB203580 group.The apoptosis of Hela299 cells was detected by Hoechst33258 assay and flow cytometry.The expression of apoptosis related proteins cleaved Caspase-3(cleaved-Caspase-3),B-cell lymphoma(Bcl-2),Bcl-2 associated X protein(Bax),and ERK/p38 MAPK protein were detected by Western blotting.Results The optimal mass concentration of CT622 protein to stimulate H ela 299 cells were 5 mg/L.CT622 significantly inhibited the apoptosis of Hela299 cells,up-regulated the expression of ERK1/2,p38MAPK and Bcl-2,and down-regulated the expression of Bax and Caspase-3 proteins.The combination of ERK1/2 and p38 MAPK inhibitors with CT622 could significantly increase the apoptosis of Hela299 cells.Conclusion T3SS effector protein CT622 inhibits Hela299 cell apoptosis by activating ERK/p38 MAPK signaling pathway.

关 键 词：沙眼衣原体 CT622效应蛋白 ERK/p38 MAPK信号通路 细胞凋亡 

分 类 号：R374.1[医药卫生—病原生物学]