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
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国家自然科学基金(81870009)


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

    Abstract:

    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.

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林少清,舒 磊,杜兴冉,冯旰珠.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[J].南京医科大学学报(自然科学版),2021,(9):1304-1309

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  • 收稿日期:2020-12-10
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  • 在线发布日期: 2021-10-09
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