Toll样受体2在结核病患者中性粒细胞凋亡中的作用及意义

The role and significance of toll-like receptor 2 in the apoptosis of neutrophils in tuberculosis patients

目的探讨Toll样受体2(TLR2)在结核病患者中性粒细胞凋亡中的作用及意义。方法分离健康成人和结核病患者外周血中性粒细胞(PMN),培养0～48 h,并于0 h、12 h、24 h和48 h用AnnexinⅤ-FITC/PI凋亡试剂盒双染、流式细胞仪检测凋亡率;应用流式细胞术检测结核病患者PMN表面TLR2表达,同时检测正常人PMN表面TLR2表达;预先加入TLR2单克隆抗体作用PMN 30 min后培养24h和48 h,用流式细胞术检测PMN凋亡率。结果结核病患者和健康成人PMN凋亡率均随时间延长而增加(P<0.05),结核病患者0～24 h凋亡率均高于健康成人PMN凋亡率,差异具有统计学意义(P<0.05);结核病患者外周血PMN表面TLR2表达高于健康成人,差异具有统计学意义(P<0.05);加入TLR2单克隆抗体预处理后,结核病患者和健康成人PMN培养24 h和48 h凋亡率均降低。结论 TLR2参与结核病患者PMN凋亡过程,阻断TLR2可降低结核病患者PMN凋亡率。

Objective To explore the role and significance of toll-like receptor 2 in neutrophil apoptosis in tuberculosis patients.Methods The fresh PMNs were isolated from peripheral blood of TB patients and health adults,cultured for 0-48 hours and stained with annexin V-FITC/PI at the time points 0 h,12 h,24 h and 48 h.The apoptosis rate of PMN was detected by flow cytometry(FCM).Meanwhile,healthy adults were used as the control group.The expressions of TLR2 on neutrophils in tuberculosis patients and healthy adults were determined respectively by flow cytometry.The fresh isolated PMNs were cultured for 24 to 48 hours with or without pretreatment of the TLR2 monoclonal antibody for 30 minutes,the apoptosis rate of PMNs was detected by flow cytometry.Results The apoptosis rate of PMNs in both healthy adults and TB patients increased with the culture time going on(P<0.05).The apoptosis rate of tuberculosis patients from 0 to 24 h was higher than that of healthy adults,and the difference was statistically significant(P < 0.05).The expression of TLR2 on PMN surface of peripheral blood of tuberculosis patients was higher than that of healthy adults,and the difference was statistically significant(P<0.05).With the addition of TLR2 monoclonal antibody pretreatment,the apoptosis rate decreased in patients with tuberculosis and healthy adults at the time points 24 h and 48 h.Conclusions TLR2 is involved in the apoptosis process of PMN in tuberculosis patients.Blocking TLR2 could reduce the apoptosis rate of PMN in tuberculosis patients.