Matsuda Takeaki, Saito Hideaki, Fukatsu Kazuhiko, Han Ilsoo, Inoue Tomomi, Furukawa Satoshi, Ikeda Shigeo, Hidemura Akio, Kang Woodae
Department of Surgery, The University of Tokyo, Japan.
Shock. 2002 Jul;18(1):69-74. doi: 10.1097/00024382-200207000-00013.
Antibiotic therapy is an essential treatment for gram-negative bacterial infections. Antibiotic-induced endotoxin release and subsequent production of inflammatory cytokines reportedly depend on the type of antibiotic action. This study examined the effects of various beta-lactam antibiotics on cell death of human polymorphonuclear neutrophils (PMNs) cocultured with Escherichia coli (E. coli) in vitro. E. coli morphology after antibiotic treatment was determined. PMNs and E. coli were cocultured with antibiotics for 0, 4, or 12 h. Levels of endotoxin and cytokines (TNF-alpha, IL-1beta, and IL-6) in the supernatants were measured. The filtrates of antibiotic-treated E. coli supernatants were cocultured with PMNs for 0, 4, or 12 h. In all experiments, ampicillin (ABPC), cefazolin sodium (CEZ), cefoperazone sodium (CPZ), latamoxef sodium (LMOX), imipenem (IPM), and polymyxin B sulfate (PLB) were used at 30 microg/mL. PMNs were isolated from healthy volunteers. PMN cell death was assessed by flow cytometry and light microscopy. ABPC, CEZ, CPZ, and LMOX, which induce bacterial filament formation with lysis, caused PMN necrosis when cocultured with E. coli. In contrast, IPM, which induces bacterial spheroplast formation with lysis, caused PMN apoptosis. Levels of endotoxin, TNF-alpha and IL-6 in the supernatants with IPM and PLB were significantly lower than in those with other beta-lactam antibiotics. The filtrates of IPM- and PLB-treated E. coli supernatants induced PMN apoptosis, whereas those treated with other beta-lactam antibiotics increased PMN necrosis. Beta-lactam antibiotics have different impacts on the types of PMN cell death after E. coli killing. Underlying mechanisms and the clinical relevance of IPM-induced PMN apoptosis in severe gram-negative infection warrant further investigation.
抗生素治疗是革兰氏阴性菌感染的重要治疗方法。据报道，抗生素诱导的内毒素释放及随后炎性细胞因子的产生取决于抗生素的作用类型。本研究检测了多种β-内酰胺类抗生素对体外与大肠杆菌（E. coli）共培养的人多形核中性粒细胞（PMN）细胞死亡的影响。确定了抗生素处理后大肠杆菌的形态。PMN和大肠杆菌与抗生素共培养0、4或12小时。测量上清液中内毒素和细胞因子（TNF-α、IL-1β和IL-6）的水平。将抗生素处理的大肠杆菌上清液的滤液与PMN共培养0、4或12小时。在所有实验中，氨苄西林（ABPC）、头孢唑林钠（CEZ）、头孢哌酮钠（CPZ）、拉氧头孢钠（LMOX）、亚胺培南（IPM）和硫酸多粘菌素B（PLB）的使用浓度均为30μg/mL。从健康志愿者中分离出PMN。通过流式细胞术和光学显微镜评估PMN细胞死亡情况。ABPC、CEZ、CPZ和LMOX可诱导细菌形成丝状并裂解，与大肠杆菌共培养时会导致PMN坏死。相比之下，IPM可诱导细菌形成原生质球并裂解，导致PMN凋亡。IPM和PLB处理的上清液中内毒素、TNF-α和IL-6的水平显著低于其他β-内酰胺类抗生素处理的上清液。IPM和PLB处理的大肠杆菌上清液的滤液可诱导PMN凋亡，而其他β-内酰胺类抗生素处理的滤液则会增加PMN坏死。β-内酰胺类抗生素在杀死大肠杆菌后对PMN细胞死亡类型有不同影响。IPM诱导PMN凋亡在严重革兰氏阴性感染中的潜在机制和临床相关性值得进一步研究。
