Viili多糖对巨噬细胞RAW264.7激活及分泌细胞因子的影响

探讨viili胞外多糖(Viili exopolysaccharides,VEPS)对小鼠巨噬细胞RAW264.7激活和增殖的影响。噻唑蓝(MTT)比色法检测细胞的生长与增殖;中性红吞噬实验检测吞噬活性;Griess试剂盒检测培养上清液中NO分泌量,ELISA法检测VEPS不同浓度及不同作用时间培养上清中IL-6,IL-1β含量;扫描电子显微镜观察VEPS对细胞形态的影响;碘化丙啶(PI)染色检测VEPS对细胞周期的影响。结果显示,VEPS对RAW264.7细胞的增殖、吞噬能力、分泌NO、IL-6、IL-1β等都有显著的促进作用,VEPS为100μg/ml时促进作用最明显,呈剂量相关,作用72h时细胞因子分泌量达到最大,72h后下降。VEPS激活巨噬细胞并使其变得扁平伸展且形成伪足。VEPS促进G1期细胞增多,提高细胞的增殖能力。VEPS免疫调节作用与其激活RAW264.7细胞,促进NO、IL-6、IL-1β等分泌有关,且VEPS与LPS对RAW264.7细胞有相似的作用规律。以上结果证明VEPS能激活巨噬细胞,也可能最终激活淋巴细胞,达到增强非特异性和特异性免疫的作用。

Effect of Viili exopolysaccharides on the activation,cell proliferation and cytokine production of murine macrophage cell line RAW264.7

This study aims to study the activation of RAW264.7 cells upon stimulation of viili exopolysaccharides(VEPS).Cell proliferation in response to different concentrations of VEPS was analyzed by MTT,phagocytosis was detected by neutral red.The secretion of,IL-6 and IL-1β in the supernatant of cell culture was measured by Griess kit and Elisa kit,respectively.Cell morphology was observed by scanning electronic microscopy(SEM),and cell cycles by flow cytometry.VEPS at 25 through 100(g/ml promoted cell proliferation,phagocytosis as evidenced by uptake of neutral red,and release of NO,IL-6 and IL-1β by RAW264.7 cells.The RAW264.7 cells treated with LPS or VEPS alone,or in combination became flattened demonstrated by SEM,a strong indication of activation of macrophages.VEPS also increased cell number in phase G1.VEPS promoted activation of macrophages in which NO,IL-1β,IL-6 were involved in a very similar way as LPS.VEPS activate macrophages and may eventually activates lymphocytes as well,and increases both innate and specific immunity.