肺炎链球菌溶血素诱导THP-1细胞凋亡涉及MAPK1/3信号途径  

Pneumolysin induces apoptosis in human THP-1 cells through MAPK1/3 signal pathway

作　　者：姜慧[1] 贺潇[1] 袁军[1] 王虹[1] 李忱炜[1] 董杰[1] 崔瑾[1] 董姗姗[1] 周爱娥[1] 张雪梅[1] 胥文春[1] 尹一兵[1] 何於娟[1] 

机构地区：[1]重庆医科大学检验系临床检验诊断学教育部重点实验室,重庆400016

出　　处：《第三军医大学学报》2011年第13期1338-1341,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金面上项目(30872363)~~

摘　　要：目的探讨肺炎链球菌溶血素(pneumolysin,Ply)对人急性单核细胞白血病单核细胞株THP-1细胞的增殖抑制和诱导凋亡作用及机制。方法 MTT法检测Ply对THP-1细胞的增殖抑制。瑞氏染色观察Ply对人THP-1细胞形态的影响。AnnexinV-FITC/PI双标法检测细胞凋亡。琼脂糖凝胶电泳观察凋亡细胞DNA片段化现象。免疫组化和Western blot检测MAPK1/3蛋白表达的变化。结果将人THP-1细胞分别与0.05、0.1、0.5、1、2.5、5μg/ml Ply共同孵育后,在24、48、72、96 h用MTT法检测细胞增殖抑制率,发现Ply对人THP-1细胞具有明显的增殖抑制作用,并且呈剂量和时间依赖性,IC50(24 h)为1.31μg/ml;0.05μg/ml和0.1μg/ml Ply分别处理THP-1细胞12 h后,通过瑞氏染色法观察人THP-1细胞可见凋亡小体及核碎裂象等典型的凋亡形态学改变;用0.05μg/ml Ply和0.1μg/ml Ply作用THP-1细胞12 h后细胞早期凋亡率分别为11.83%和48.45%(P<0.05);提取凋亡THP-1细胞DNA,进行琼脂糖凝胶电泳可见典型的"梯状"条带;Ply作用于人THP-1细胞后,胞内MAPK1/3蛋白表达降低(P<0.05)。结论 Ply可诱导人THP-1细胞凋亡,此作用涉及MAPK1/3信号通路的参与。Objective To determine the effect of pneumolysin（Ply） on proliferation and apoptosis in human acute monocytic leukemia THP-1 cells.Methods After exposure to Ply at 0.05,0.1,0.5,1,2.5,or 5 μg/ml for 24,48 or 72 h,the proliferation in THP-1 cells was measured by MTT assay.Morphological changes in the treated and untreated THP-1 cells were observed by Wright＇s staining.The apoptotic rate of THP-1 cells was examined by AnnexinV-FITC/PI-stain.DNA fragmentation was examined by sepharose electrophoresis.The expression of MAPK1/3 was investigated by immunocytochemical staining and Western blot analysis.Results Ply resulted in apparent inhibition on proliferation of THP-1 cells in a time-and dose-dependent manner,with an IC50（24 h） of 1.31 μg/ml.Typical morphological features of apoptosis,such as nuclear fragmentation and apoptosis bodies,were observed in THP-1 cells after Ply treatment at 0.05 and 0.1 μg/ml for 12 h by Wright＇s staining.After Ply treatment at 0.05 and 0.1 μg/ml for 12 h,the apoptotic rate of THP-1 cells was 11.83% and 48.45%,respectively（P〈0.05）.A typical ladder pattern was identified in DNA electrophoresis.The expression of MAPK1/3 was reduced（P〈0.05） after THP-1 cells were treated with Ply.Conclusion Ply induces apoptosis in human THP-1 cells though MAPK1/3 signal pathway.

关 键 词：肺炎链球菌溶血素 THP-1细胞 凋亡 MAPK1/3通路 

分 类 号：R378.18[医药卫生—病原生物学] R733.71[医药卫生—基础医学]