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[铜绿假单胞菌弹性蛋白酶抑制肺表面活性蛋白A介导的免疫吞噬作用]

[Pseudomonas aeruginosa elastase inhibits immune phagocytosis mediated by pulmonary surfactant protein A].

作者信息

Gao Yang, Wang Zeng, Wang Junqin, Cai Guodong, Zhu Zhuangchen, Chen Guang

机构信息

Department of Orthopaedics, Affiliated Hospital of Taishan Medical College, Taian 271000, China.

出版信息

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2014 Dec;30(12):1251-4.

Abstract

OBJECTIVE

To investigate the relationship between Pseudomonas aeruginosa (P.aeruginosa) elastase and pulmonary surfactant protein A (SP-A) in host infected by P.aeruginosa.

METHODS

C3H mice were intranasally infected with P.aeruginosa wild-type PAO1, ΔlasB mutant and genetic complement strain PDO240LasB to determine the difference of virulence between wide type and mutant. The ability to degrade SP-A in vitro by PAO1, ΔlasB and PDO240LasB was observed through co-incubation of equal bacteria and SP-A and detected by Western blotting. The susceptibility of bacteria to phagocytosis was assayed by in vitro experiment that bacteria treated with SP-A was incubated with mouse Raw264.7 macrophages.

RESULTS

Compared with wide-type PAO1, the virulence of ΔlasB mutant was attenuated in the mouse model of P.aeruginosa infection because of the knock down of elastase expression. The ΔlasB mutant lost the ability to degrade SP-A when incubated with SP-A in vitro. The in vitro phagocytosis experiments showed that SP-A augmented the phagocytosis of ΔlasB mutant bacteria more efficiently than the wild-type PAO1.

CONCLUSION

P.aeruginosa elastase provides a protection from phagocytic cells by degrading SP-A.

摘要

目的

研究铜绿假单胞菌弹性蛋白酶与铜绿假单胞菌感染宿主时肺表面活性蛋白A(SP-A)之间的关系。

方法

用铜绿假单胞菌野生型PAO1、ΔlasB突变体和基因互补菌株PDO240LasB经鼻感染C3H小鼠,以确定野生型和突变体之间的毒力差异。通过等量细菌与SP-A共孵育,观察PAO1、ΔlasB和PDO240LasB在体外降解SP-A的能力,并通过蛋白质免疫印迹法进行检测。通过将经SP-A处理的细菌与小鼠Raw264.7巨噬细胞共孵育的体外实验,测定细菌对吞噬作用的敏感性。

结果

与野生型PAO1相比,由于弹性蛋白酶表达下调,ΔlasB突变体在铜绿假单胞菌感染小鼠模型中的毒力减弱。ΔlasB突变体在体外与SP-A共孵育时丧失了降解SP-A的能力。体外吞噬实验表明,SP-A增强对ΔlasB突变体细菌的吞噬作用比野生型PAO1更有效。

结论

铜绿假单胞菌弹性蛋白酶通过降解SP-A为吞噬细胞提供保护。

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