骆驼蓬多糖以自噬途径免疫调节炎症细胞模型
目的：以骆驼蓬多糖干预小鼠腹腔巨噬细胞RAW264.7炎症反应模型,探讨骆驼蓬多糖调节炎症反应的生物活性及意义。方法：利用稳定表达GFP-LC3的NRK细胞,用不同浓度的骆驼蓬多糖干预,诱导细胞自噬,用激光共聚焦显微镜进行观察自噬体,筛选出骆驼蓬多糖诱导细胞自噬的最佳干预浓度和时间范围；利用RAW264.7细胞,以1μg/mL脂多糖构建炎症反应细胞模型；参照骆驼蓬多糖诱导细胞自噬的最佳浓度和时间进行干预,以CCK-8试验检测细胞活性,选择Hoechst和PI双重染色方法观察细胞凋亡,用ELISA试剂检测炎症因子IL-1β和TNF-α的分泌量。结果：骆驼蓬多糖对NRK细胞诱导自噬的最佳时间为12h,浓度为50μg/mL-100μg/mL范围。骆驼蓬多糖干预LPS诱导的RAW264.7细胞炎症模型后,抑制细胞增殖活性,诱导细胞凋亡,呈剂量依赖性,细胞培养液中,TNF-a和IL-1β分泌量降低,与模型组有显著差异(p<0.05)。结论：骆驼蓬多糖能抑制LPS刺激诱导的巨噬细胞增殖,诱导细胞自噬和凋亡,能抑制LPS诱导的炎症因子。
Objective: The aim of this study was to investigate the biological activity and significance of inflammatory response and the Potential mechanism of Peganum harmala polysaccharides on LPS－induced inflammatory response in RAW264.7cells . Methods: The NRK cells that stably expressing GFP-LC3 were treated by different concentrations of Peganum harmala polysaccharide,and Autophagy was induced and autophagosomes were observed by laser confocal microscopy. The optimal concentration and time range of autophagy induced by Peganum harmala polysaccharide were screened. 1ug/mL,Lipopolysaccharide was applied to RAW264.7 cells to establish an inflammatory Cell model and According to the above concentration range Treated with polysaccharides,The cells viability was measured by CCK-8,Using Hoechst and PI double staining method to observe apoptosis under laser confocal microscopy. The secretion of inflammatory factors IL-1βand TNF-α was detected by enzyme-linked immunosorbent assay (ELISA). Results:The optimal time for autophagy induced by Peganum harmala polysaccharide to NRK cells was 12 h, and the concentration ranged from 50μg/mLto100μg/mL.RAW264.7cell inflammatory mode treated by The Peganum harmala polysaccharide and inhibited cell proliferation activity and induced apoptosis in a dose-dependent manner,The secretion of TNF-a and IL-1β was decreased in cell culture medium, which was significantly different from the model group p<0.05. Conclusion: Peganum harmala polysaccharide can inhibit the proliferation of macrophages induced by LPS, induce autophagy and apoptosis, and inhibit the inflammatory factors induced by LPS.
标题：骆驼蓬多糖以自噬途径免疫调节炎症细胞模型
英文标题：Immunomodulatory Effect of Peganum harmala polysaccharide On the inflamatory cell model by autophagy
作者：
曼则热姆.热杰普 新疆医科大学基础医学院生物化学教研室
木塔力甫.艾买提 新疆医科大学中心实验室
伊力亚斯.艾萨 新疆医科大学基础医学院生物化学教研室
冯学召 新疆医科大学基础医学院生物化学教研室
古丽努尔.阿不力米提 新疆医科大学基础医学院生物化学教研室
雷秀英 新疆医科大学基础医学院生物化学教研室
陈倩 新疆医科大学基础医学院生物化学教研室
米娜 新疆医科大学基础医学院生物化学教研室;新疆医科大学中心实验室;新疆医科大学基础医学院生物学教研室;新疆医科大学基础医学院生物学教研室
中文关键词:小鼠腹腔巨噬细胞RAW264.7,骆驼蓬多糖,LPS,IL-1β,TNF-α,
英文关键词:Mouseperitoneal macrophage RAW264.7,Peganum harmala polysaccharides,LPS,IL-1β,TNF-α,
发表日期：2018-12-28
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