结核分枝杆菌19 kDa脂蛋白诱导THP-1细胞凋亡的研究

Study on apoptosis of THP-1 cells induced by Mycobacterium tuberculosis 19 kDa lipoprotein

摘要: 目的:观察结核分枝杆菌19 kDa脂蛋白(Mtb P19)对THP-1细胞凋亡的影响。方法:从培养的结核杆菌H37Ra制备Mtb P19,以5、10、20μg/ml Mtb P19作用于佛波醇酯(PMA)分化的THP-1细胞,37℃、5% CO₂孵箱孵育4 h。Wright-Giemsa染色观察细胞形态变化;AnnexinV-FITC染色,用流式细胞仪检测细胞凋亡。结果:终浓度为5μg/ml的Mtb P19即可诱导THP-1细胞凋亡,且凋亡率随Mtb P19作用浓度的升高而增加(P<0.01);Wright-Giemsa染色镜下可见部分细胞体积变小、圆缩,核固缩,核断裂。结论:结核分枝杆菌P19以剂量依赖方式诱导THP-1细胞凋亡。

Abstract: Objective: To abserve the influence of Mycobacterium tuberculosis 19 kDa(Mtb P19) on the apoptosis of THP-1 cells.Methods: Mtb P19 was prepared from cultured Mtb H37Ra.Phorbol myristate acetate-differentiated THP-1 cells were incubated with P19 at the concentrations from 5 μg/ml to 20 μg/ml at 37℃,5% CO₂ for up to 4 hours.The change of cells was observed by Wright-Giemsa stain,and the apoptosis rate was detected by flowcytometry.Results: Mtb P19 could induce THP-1 cells apoptosis at final concentration of 5 μg/ml,and the apopotosis rate increased with the concentration of P19(P<0.01).Round shrinkage,pyknosis or nuclear fragmentation were observed in some cells by Wright-Giemsa stain.Conclusions: Mtb P19 induces apoptosis in THP-1 cells in a dose-dependent manner.