摘要
目的:观察表面活性蛋白A(SP-A)对大肠杆菌凝集、生长的作用及单核巨噬细胞对大肠杆菌摄取的影响,探讨SP-A在固有免疫中的作用及机制。方法:①SP-A对大肠杆菌凝集的影响:培养大肠杆菌至稳定期,进行分组:空白对照组、BSA组(BSA 25mg/L)、SP-A组(SP-A 25mg/L)。37℃,200r/min培养,观察大肠杆菌形态。②SP-A对单核巨噬细胞吞噬能力的影响:Ficoll-Hypaque密度梯度离心法分离健康志愿者外周血,分离单核巨噬细胞。实验分为空白对照组、SP-A组(SP-A 25mg/L)。快速瑞氏-姬姆萨复合染液染色,显微镜下观察单核巨噬细胞对大肠杆菌的吞噬并摄像。③SP-A对大肠杆菌生长的影响:培养大肠杆菌至稳定期,实验分为空白对照组、BSA组(BSA 25mg/L)、SP-A组(SP-A 25mg/L)。37℃,200r/min培养6h,每1h末于600nm波长分光光度计进行光密度值(OD)测定,绘制大肠杆菌生长曲线。结果:①空白对照组、BSA组大肠杆菌未发生凝集,SP-A组大肠杆菌发生凝集。②与空白对照组相比,加入SP-A后单核巨噬细胞对大肠杆菌摄取明显增加。③加入BSA后,大肠杆菌生长无明显变化(P>0.05);加入SP-A后,大肠杆菌生长受到明显抑制(P<0.05)。结论:SP-A通过促进大肠杆菌凝集发生,一方面直接抑制大肠杆菌生长;另一方面增加单核巨噬细胞对大肠杆菌的摄取,从而在固有免疫中发挥作用。
Objective： To investigate the role and its mechanism of surfactant protein A on Escherichia coli aggregation, growth and phagocytosis of macrophage. Methods： ① The role of surfactant protein A on Escherichia coli aggregation： Escherichia coli were grown at 37°C in LB medium with BSA（25 mg/L）, SP-A（25 mg/L）. Escherichia coli aggregation was evaluated under the light microscope after 1-hour culture. ②The role of surfactant protein A on phagocytosis of macrophage： Macrophage was isolated from healthy peripheral venous blood with Fieoll-Hypaque density gradient centrifugation. The macrophage were exposed to medium with SP-A （25 mg/L） and the number of Phagocyted Escherichia coli in macrophage was counted under microscope. ③The role of surfactant protein A on Escherichia coli aggregation： Escherichia coli were grown at 37°C in LB medium with BSA（25 mg/L） ,SP-A（25 mg/L）. Escherichia coli density was analysed by a spectrophometer at a wavelength of 600 nm at one hour intervals. Results： ①Aggregated-Escherichia coli were founded in SP-A medium, which was not in control mediums （P〈0.05）. ②Growth of Escherichia coli SP-A addition significantly increased controls. Conclusion： SP-A plays an was inhibited in SP-A medium, but not in BSA medium. ③the number of Escherichia coli in macrophage in contrast to lmpo eration of Escherichia coli and improving rtant role in immunity via inhibiting the ability of proliferation of Escherichia coli and macrophage-induced phagocytosis of Escherichia coli.
出处
《武汉大学学报（医学版）》
CAS
北大核心
2014年第2期202-204,208,共4页
Medical Journal of Wuhan University
基金
国家自然科学基金资助项目(编号:30670985
81070556)
