铜绿假单胞菌分泌蛋白Pec1抑制肺巨噬细胞吞噬功能的初步研究  

Inhibitory effect of Pseudomonas aeruginosa secreted protein Pec1 on phagocytosis of pulmonary alveolus macrophages

作　　者：林少清 舒磊 杜兴冉[2] 冯旰珠[1] LIN Shaoqing;SHU Lei;DU Xingran;FENG Ganzhu(Department of Respiratory and Critical Care Medicine,the Second Affiliated Hospital of Nanjing Medical University,Nanjing 210011,China;Department of Infectious Disease Department,the Second Affiliated Hospital of Nanjing Medical University,Nanjing 210011,China)

机构地区：[1]南京医科大学第二附属医院呼吸与危重症医学科,江苏南京210011 [2]南京医科大学第二附属医院感染科,江苏南京210011

出　　处：《南京医科大学学报（自然科学版）》2021年第9期1304-1309,共6页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金(81870009)。

摘　　要：目的:探讨铜绿假单胞菌分泌蛋白Pec1对小鼠肺泡巨噬细胞株MH-S吞噬功能的影响。方法:通过PCR扩增、质粒构建、原核表达及蛋白纯化等过程制备重组蛋白Pec1;CCK-8法检测Pec1蛋白对MH-S细胞增殖的影响;中性红染色检测MH-S细胞的吞噬功能;荧光显微镜观察MH-S细胞对铜绿假单胞菌标准菌株PAO1灭活菌的吞噬能力。结果:成功构建重组质粒pET-30a-pec1,原核表达并纯化后得到重组蛋白Pec1,其分子量为30.5 kDa。Pec1对MH-S细胞的增殖有抑制作用,Pec1浓度越高,对细胞增殖的抑制作用越明显;Pec1能减少MH-S细胞对中性红的内吞,抑制MH-S细胞对铜绿假单胞菌灭活菌的吞噬作用。结论:铜绿假单胞菌分泌蛋白Pec1对MH-S细胞的吞噬功能有抑制作用。Objective:This study aims to investigate the effect of the Pseudomonas aeruginosa secreted protein Pec1 on phagocytosis of MH-S cells.Methods:The recombinant protein Pec1 was prepared by PCR amplification,plasmid construction,induced expression and protein purification.The effect of the recombinant protein Pec1 on the proliferation of MH-S cells was measured by CCK-8 kit.Neutral red was used to detect the phagocytic function of MH-S cells,and the phagocytosis of heat-inactivated Pseudomonas aeruginosa was observed by fluorescence microscope.Results:The recombinant protein Pec1 with the molecular weight of 30.5 kDa was successfully expressed and purified.Pec1 had a concentration-dependent inhibitory effect on the proliferation of MH-S cells.Pec1 decreased the endocytosis of neutral red and inhibited the phagocytosis of heat-inactivated Pseudomonas aeruginosa by MH-S cells.Conclusion:The Pseudomonas aeruginosa secreted protein Pec1 can inhibit the phagocytosis of MH-S cells.

关 键 词：Pec1 铜绿假单胞菌 吞噬作用 

分 类 号：R392.12[医药卫生—免疫学]