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目的探讨表面活性蛋白D(SP-D)对吞噬细胞吞噬幽门螺杆菌(HP)的影响。方法从SD大鼠肺泡灌洗液中提取并纯化SP-D。将试验组HP11637、HPSS1菌液分别用SP-D预处理30min,加入小鼠腹腔巨噬细胞培养液。将细胞与细菌混合物置于细胞培养箱中37℃分别孵育0、30、60min后,用头孢唑啉钠杀灭细胞外的细菌。裂解巨噬细胞后,取适量细胞裂解稀释液接种于微需氧环境中孵育3d后,计数菌落形成单位。同时设置未经SP-D预处理的HP11637、HPSS1加入小鼠腹腔巨噬细胞培养液中作为对照组。结果小鼠腹腔巨噬细胞与HP11637共同孵育后0、30、60min,试验组细胞内HP11637数量为(123±8)、(411±38)、(343±44)个/100个巨噬细胞,显著多于对照组的(104±14)、(179±10)、(163±10)个/100个巨噬细胞(P<0.01)。小鼠腹腔巨噬细胞与HPSS1共同孵育后0、30、60min,两组细胞内HPSS1数量相仿(P>0.05)。结论 SP-D具有显著增强小鼠腹腔巨噬细胞体外吞噬HP11637的功能,而对小鼠腹腔巨噬细胞体外吞噬HPSS1无显著影响。
Objective To investigate the effect of surfactant protein D (SP-D) on phagocytosis of Helicobacter pylori (HP). Methods SP-D was extracted and purified from BAL fluid in BALF. The experimental group HP11637, HPSS1 bacteria were pretreated with SP-D for 30min, added to the peritoneal macrophage culture medium. The cells and bacteria mixture were placed in a cell incubator at 37 ℃ were incubated 0,30,60 min, the use of cefazolin sodium to kill extracellular bacteria. After lysing macrophages, take appropriate cell lysis diluent was inoculated in micro-aerobic environment incubated 3d, colony forming units. Meanwhile, HP11637 and HPSS1 without SP-D pretreatment were added into the culture medium of mouse peritoneal macrophages as a control group. Results The number of intraperitoneal HP11637 was (123 ± 8), (411 ± 38) and (343 ± 44) / 100 macrophages in the experimental group at 0, 30, and 60 minutes after co-incubation with mouse peritoneal macrophages. (104 ± 14), (179 ± 10), (163 ± 10) / 100 macrophages in the control group (P <0.01). Mice peritoneal macrophages co-incubated with HPSS1 0,30,60 min, the number of intracellular HPSS1 similar (P> 0.05). Conclusion SP-D can significantly enhance the function of murine peritoneal macrophages in phagocytosing HP11637 in vitro and have no significant effect on in vitro phagocytosis of HPSS1 induced by peritoneal macrophages in mice.