硒化大蒜多糖对小鼠腹腔巨噬细胞功能的影响

【目的】 研究硒化大蒜多糖（sGPS）对小鼠腹腔巨噬细胞功能的影响，以期为硒化大蒜多糖的作用挖掘和临床应用提供依据。【方法】 依次通过分离、纯化得到大蒜多糖（GPS），并经硝酸-亚硒酸钠硒化修饰得到sGPS₃、GPS₅和sGPS₆。以小鼠腹腔巨噬细胞为研究对象，用6.25、12.5、25、50、100 μg/mL sGPS₃、GPS₅、sGPS₆及10 μg/mL脂多糖（LPS组）处理小鼠腹腔巨噬细胞48 h，同时设置不加药物的细胞为对照组，用中性红法测定其吞噬功能，CCK-8法测定其増殖能力，筛选出活性最好的硒化大蒜多糖；然后用ELISA法检测活性最好的硒化大蒜多糖对巨噬细胞上清液中一氧化氮（NO）、肿瘤坏死因子-α（TNF-α）、干扰素-γ（IFN-γ）、白介素-1β（IL-1β）、白介素-6（IL-6）、白介素-12（IL-12）含量的影响；再通过小鼠碳廓清实验、小鼠腹腔巨噬细胞吞噬鸡红细胞实验测定空白对照组（生理盐水）及高（2 mg/mL）、中（1 mg/mL）、低（0.5 mg/mL）剂量活性最好的硒化大蒜多糖的吞噬指数与吞噬百分率。【结果】 除6.25 μg/mL sGPS₃外，6.25、12.5、25、50和100 μg/mL sGPS₃、sGPS₅、sGPS₆组小鼠腹腔巨噬细胞的A_{490 nm}值均显著高于对照组（P<0.05），其中，50和100 μg/mL sGPS₆组的A_{490 nm}值显著高于LPS组（P<0.05），因此选用sGPS₆进行后续试验；12.5、25、50和100 μg/mL sGPS₆组小鼠腹腔巨噬细胞上清中NO、TNF-α、IL-1β、IL-6含量均显著高于对照组，25、50和100 μg/mL sGPS₆组小鼠腹腔巨噬细胞上清中IFN-γ、IL-12显著高于对照组（P<0.05），其中100 μg/mL sGPS₆组小鼠腹腔巨噬细胞上清中NO、TNF-α、IL-12显著高于LPS组（P<0.05）。2和1 mg/mL sGPS₆组的吞噬指数和吞噬率均显著高于空白对照组（P<0.05）。【结论】 硒化大蒜多糖能增强小鼠腹腔巨噬细胞的吞噬功能和増殖能力，促进细胞因子TNF-α、IL-1β和IL-6的分泌。

Effects of Selenized Garlic Polysaccharide on the Function of Mouse Peritoneal Macrophages

【Objective】 This study was aimed to explore the effect of selenized garlic polysaccharide on the function of mouse peritoneal macrophages, in order to provide a basis for the mining and clinical application of selenium garlic polysaccharide (sGPS).【Method】 The garlic polysaccharide(GPS) was obtained by isolation and purification, and modified by HNO₃-Na₂SeO₃ to obtain sGPS₃, GPS₅ and sGPS₆.Taking the mouse peritoneal macrophages as the research object, the mouse peritoneal macrophages were treated with sGPS₃, GPS₅, sGPS₆ at five concentrations (6.25, 12.5, 25, 50 and 100 μg/mL)and 10 μg/mL lipopolysaccharide (LPS group) for 48 h, at the same time, the cells without drugs were set as control group. The neutral red method was used to determine the phagocytic function of mouse peritoneal macrophages, and the CCK-8 method was used to determine the proliferation, in order to screen out sGPS with the best activity. The ELISA method was used to determine the effect of sGPS with the best activity on the contents of NO, TNF-α, IFN-γ, IL-1β, IL-6, and IL-12 in the supernatant of macrophages. The mouse peritoneal macrophages swallow chicken red blood cells and mouse carbon clearance experiments were used to detemine the phagocytic indexes and phagocytic percentage of blank control group (normal saline), high (2 mg/mL), medium (1 mg/mL) and low (0.5 mg/mL) doses of sGPS with the best activity.【Result】 Except 6.25 μg/mL sGPS₃, the A_{490 nm} values of 6.25, 12.5, 25, 50 and 100 μg/mL peritoneal macrophages in sGPS₃, sGPS₅ and sGPS₆ groups were significantly higher than those in control group (P<0.05).Among them the A_{490 nm} values 50 and 100 μg/mL of sGPS₆ groups were significantly higher than that of the LPS group (P<0.05).Therefore, sGPS₆ was selected for follow-up test.The contents of NO, TNF-α, IL-1β and IL-6 in the supernatant of mouse peritoneal macrophages of sGPS₆ at the concentrations of 12.5, 25, 50 and 100 μg/mL were significantly higher than those in control group (P<0.05).The contents of IFN-γ, IL-12 in the supernatant of mouse peritoneal macrophages of sGPS₆ at the concentrations of 25, 50 and 100 μg/mL were significantly higher than those in control group (P<0.05).Among them, the contents of NO, TNF-α and IL-12 of mouse peritoneal macrophage supernatant at 100 μg/mL of sGPS₆ were significantly higher than the LPS group (P<0.05).The phagocytic index and phagocytic rate at 2 and 1 mg/mL sGPS6 were significantly higher than those of the blank control group (P<0.05).【Conclusion】 Selenium garlic polysaccharide could enhance the phagocytic function and proliferation ability of the mouse peritoneal macrophages, and promote the secretion of cytokines TNF-α, IL-1β and IL-6.