金黄色葡萄球菌毒力因子PSM-α对巨噬细胞的细胞毒性和呼吸爆发的影响
首发时间:2019-08-13
摘要:α型酚溶性调节蛋白(PSM-α)是金黄色葡萄球菌产生的重要毒力因子,可以高效裂解巨噬细胞。在本研究中,我们通过连接基因PSM-α与表达载体pGEX-4T-1,成功构建了重组质粒pGEX-PSM-α。IPTG诱导大肠杆菌DE3蛋白表达,通过纯化和凝血酶酶切获得纯化蛋白。纯化蛋白刺激分离鼠腹腔巨噬细胞,通过流式细胞术以及CCK-8鉴定PSM-α对巨噬细胞细胞毒性和呼吸爆发的影响。结果表明,PSM-α可以显著提高巨噬细胞的呼吸爆发。因此得出结论,PSM-α可激活巨噬细胞呼吸爆发的发生,可能是金黄色葡萄球菌临床治疗中潜在的抗炎药物靶标。
关键词: 金黄色葡萄球菌 PSM-α 串联融合表达 巨噬细胞
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Effect of Staphylococcus aureus virulence factor PSM-α on cytotoxicity and respiratory burst of Macrophages
Abstract:Phenol-soluble modulin α-type (PSM-α) is an important virulence factor secreted by Staphylococcus aureus (S. aureus), which has the ability to lyse macrophage. In this study, we connected PSM-α with the expression vector pGEX-4T-1 to construct recombinant plasmids pGEX-PSM-α. Then, we uesd Escherichia coli DE3 to expressed PSM-α, and got its soluble protein through purification and thrombin digestion. Then, we stimulated mouse peritoneal macrophage with purified protein. We detected the impact on macrophage by Flow Cytometry and CCK-8. The results showed that PSM-α can significantly increase the respiratory burst of macrophages. Therefore, it is concluded that PSM-α can activate the occurrence of macrophage respiratory bursts, which may be potential anti-inflammatory drug targets in the clinical treatment of Staphylococcus aureus
Keywords: Staphylococcus aureus PSM-α tandem fusion expression macrophage
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金黄色葡萄球菌毒力因子PSM-α对巨噬细胞的细胞毒性和呼吸爆发的影响
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