LL-37抗菌肽提纯及其治疗革兰阴性细菌感染的研究  

Purification of LL-37 Peptide and Its Effects on Gram-negative Bacterial Infection

作　　者：冯新富[1] 李俊华[1] 陈晓理[2] 石承先[1] 王伯瑶[3] 

机构地区：[1]贵州省人民医院普外科,贵阳500011 [2]四川大学华西医院普外科 [3]四川大学华西基础医学院感染免疫室

出　　处：《天津医药》2006年第9期628-630,共3页Tianjin Medical Journal

基　　金：国家自然科学基金资助项目(项目编号:39970932)

摘　　要：目的:提纯抗菌肽LL-37并探讨其治疗小鼠革兰阴性(G-)细菌感染的疗效。方法:用蛋白质纯化技术从pGEX-1λT-LL-37重组质粒转染的大肠杆菌JM109中提纯LL-37肽,检测LL-37对G-细菌(铜绿假单胞菌)的杀菌活性。随机将BALB/C小鼠分配入盐水组(n=8)、内毒素组(n=8)、LL-37结合内毒素组(n=8),以观察LL-37结合内毒素的能力;随机将BALB/C小鼠分配入盐水组(n=8)、G-细菌感染组(n=8)、LL-37治疗组(n=8),以观察LL-37治疗BALB/C小鼠G-细菌感染的疗效。结果:融合蛋白和LL-37分子质量分别为30ku和5ku;LL-37对铜绿假单胞菌的最小抑菌浓度(MIC)、最小有效浓度(MEC)、最小杀菌浓度(MBC)分别为25、100、50mg/L;内毒素组、LL-37结合内毒素组48h死亡率为100%、12.5%;G-细菌感染组、LL-37治疗组48h死亡率为75%、37.5%。结论:LL-37具备杀菌、抗内毒素毒性能力,可用于治疗革兰阴性细菌感染。Objective： To purify LL-37 peptide and explore its effects on the treatment of gram-negative bacterial infection in BALB/C mice. Methods： Purified LL-37 was extracted from Escherichia coli JM 109 which were transfected by the pGEX- 1λT-LL-37 recombinant vector. The antibacterial activity of LL-37 for gram-negative bacteria （pseudomonas） was detected. BALB/C mice were divided randomly into saline group, endotoxin group and LL-37 Combined with endotoxin group （n=8）. BALB/C mice were randomly divided into saline group, gram-negative bacterial infection group, and LL-37 group. Results： The molecular magnitudes of fusion protein and LL-37 recombinant peptide were approximate 30 ku and 5 ku respectively. The minimal inhibitory concentration （M IC）, minimal effective concentration （MEC）, and minimal bactericidal concentration （MBC） of LL- 37 to pseudomonas were 25, 100, 50 mg/L respectively. The 48 hour mortality was 100% in endotoxin group, 12.5% in LL-37 combined with endotoxin group, 75% in gram-negative bacterial infection group, and 37.5% in LL-37 group. Conclusion： LL-37 has an ability of killing bacteria and attenuating the leathality of endotoxin, and can be used to treat gram-negative bacterial infection.

关 键 词：革兰阴性菌 感染 抗生素 肽 假单胞菌 铜绿 大肠杆菌 微生物敏感性试验 小鼠 

分 类 号：R978.16[医药卫生—药品] R978.1[医药卫生—药学]