中性粒细胞吞噬表皮葡萄球菌诱导自身凋亡的形态学研究

Morphological study of neutrophils in phagocytizing S.epidermidis and inducing self-apoptosis

目的 利用中性粒细胞的吞噬功能 ,进一步探索中性粒细胞在体内吞噬细菌的变化和转归问题及可能机制。方法 取静脉血 1.8ml抗凝 ,加入表皮葡萄球菌菌液 0 .2ml( 10 9/ml) ,混匀 ,37℃恒温摇床振荡培养 ,每隔 30min取一滴培养物制备血涂片 ,瑞 姬染色 ,油镜下观察中性粒细胞形态变化 ,直到第 10h。同时以生理盐水做对照组。结果 30min即可见到中性粒细胞吞噬细菌现象 ;到第 3小时 ,吞噬细菌的中性粒细胞核发生裂解 ,出现分裂丝 ,直到第 4小时 ,就可见到吞噬细菌的中性粒细胞的典型凋亡特征。同时 ,胞核呈肾形、胞体较小的单核细胞也可吞噬细菌 ,但吞噬的细菌数较少 ;胞核呈“山”形且较大的单核细胞“粘附”核刚发生裂解的中性粒细胞。结论 ( 1)被中性粒细胞吞噬的表皮葡萄球菌能够最终激活凋亡基因 ,导致中性粒细胞凋亡 ;( 2 )

Objective To investigate the possible mechanism of neutrophils in phagocytizing S.epidermidis by means of neutrophils'phagocytosis.Methods Blood 1.8 ml was obtained from vein and anticoagulated,S.epidermidis liquid (10 9/ml) 0.2 ml was added and mixed fully,then cultured in a shaking cradle at 37 ℃.One drop of cultured liquid was taken out at 30 min interval lasting for 10 hours,which was stained by Wright Giemsa.Morphological changes of neutrophils were observed under oil immersion lens,meanwhile NS served as the control group.Results Neutrophils'phagocytosis occured after 30 min;the karyorrhexis of neutrophils was observed after 3 hours;typical apoptosis characteristics could be found after 4 hours.Meanwhile,smaller monocytes whose karyons looked like a kidney could also phagocytize few bacteria,and bigger monocytes whose karyons looked like 'W' could adhere neutrophlis whose karyons were just split.Conclusion (1)The S.epidermidis phagocytized by neutrophils can eventually activate apoptosis genes and thus induce neutrophils'apoptosis;(2)The different structural features of monocytes in the blood are related to the different phagocytosis of theirs.