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[兔中性粒细胞防御素对铜绿假单胞菌及其多重耐药菌株的体外抗菌活性]

[In vitro antimicrobial activity of defensins from rabbit neutrophils against Pseudomonas aeruginosa and its multiple-drug-resistance strains].

作者信息

Zhao Wei-ye, Dong Bi-rong, Zhou Yan

机构信息

Department of Geriatrics, West China hospital of Sichuan University, Chengdu.

出版信息

Sichuan Da Xue Xue Bao Yi Xue Ban. 2005 Jan;36(1):83-5.

Abstract

OBJECTIVE

To investigate the role and mechanism of defensins from rabbit neutrophils against the multi-drug-resistant (MDR) strains of Pseudomonas aeruginosa.

METHODS

Defensins were extracted from rabbit peritoneal neutrophil granules with citric acid and purified by acid urea polyacrylamide gel electrophoresis (AU-PAGE). An agrose gel radial diffusion assay for bactericidal activity in vitro was used and ultrastructural changes of the killed Pseudomonas aeruginosa were examined by use of transmission electron microscope.

RESULTS

NP2 was isolated and purified successfully by AU-PAGE. NP2 demonstrated concentration-dependent killing of Pseudomonas aeruginosa and its MDR strain. When the two kinds of Pseudomonas aeruginosa cells exposed to NP2 were examined by transmission electron microscope, we found that the cell membrane was incomplete, the components of cells condensed into pyknotic bodies accompanied by vacuolization, and there was some transudate between the cells.

CONCLUSION

Defensin from the rabbit neutrophils exerts potent bactericidal activity against the MDR strain of Pseudomonas aeruginosa. The findings under electron microscope suggest the theory that defensins are capable of destabilizing the cell membrane permeability and causing the death of microorganism.

摘要

目的

研究兔中性粒细胞防御素对多重耐药铜绿假单胞菌菌株的作用及其机制。

方法

用柠檬酸从兔腹膜中性粒细胞颗粒中提取防御素,通过酸性尿素聚丙烯酰胺凝胶电泳(AU-PAGE)进行纯化。采用琼脂糖凝胶放射扩散法检测体外杀菌活性,并利用透射电子显微镜观察被杀灭的铜绿假单胞菌的超微结构变化。

结果

通过AU-PAGE成功分离并纯化出NP2。NP2对铜绿假单胞菌及其多重耐药菌株表现出浓度依赖性杀伤作用。当用透射电子显微镜检查两种暴露于NP2的铜绿假单胞菌细胞时,发现细胞膜不完整,细胞成分浓缩成固缩体并伴有空泡化,细胞间有一些渗出液。

结论

兔中性粒细胞防御素对多重耐药铜绿假单胞菌菌株具有强大的杀菌活性。电子显微镜下的发现提示了防御素能够破坏细胞膜通透性并导致微生物死亡的理论。

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