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目的探讨金黄色葡萄球菌(简称金葡菌)感染人巨噬细胞系U937细胞后半胱氨酸蛋白酶-3/-9(Caspase-3/-9)的表达,确定Caspase-3/-9在金葡菌诱导的U937细胞凋亡中的作用。方法当细胞:细菌为1:20时分别培养0、15、30、60和90min,采用Annexin V FITC/PI双染流式细胞仪检测U937细胞的凋亡率;采用Westernblot方法检测Caspase-3和Caspase-9的表达;采用荧光比色法检测Caspase-3/-9蛋白酶活性。结果金葡菌以时间依赖的方式诱导U937细胞凋亡。随着金葡菌感染时间的延长,Caspase-3和Cas-pase-9的表达逐渐增加,Caspase-3/-9蛋白酶活性随着培养时间的延长也逐渐上升。Caspases蛋白酶的抑制剂Z-DEVD-FMK和Z-LEHD-FMK能够抑制金葡菌诱导的U937细胞凋亡。结论Caspase-3/-9参与了金葡菌诱导的U937凋亡过程并发挥了重要的作用。
Objective To investigate the expression of Caspase-3 / -9 in human macrophage cell line U937 infected with Staphylococcus aureus (Staphylococcus aureus) Role of Staphylococcus aureus in Apoptosis Induced by U937 Cells. Methods The apoptosis rate of U937 cells was detected by Annexin V FITC / PI flow cytometry at 0, 15, 30, 60 and 90 min after the cells were 1:20, respectively. The expressions of Caspase-3 and Caspase-9 expression; Caspase-3 / -9 protease activity was detected by fluorescence colorimetry. Results Staphylococcus aureus induced U937 cell apoptosis in a time-dependent manner. With the prolongation of S. aureus infection, the expression of Caspase-3 and Cas-pase-9 gradually increased, and the activity of Caspase-3 / -9 gradually increased with the prolongation of culture time. Inhibitors of Caspases protease, Z-DEVD-FMK and Z-LEHD-FMK, inhibited S. aureus-induced apoptosis of U937 cells. Conclusion Caspase-3 / -9 is involved in the apoptosis of U937 induced by S. aureus and plays an important role.