基于LPS炎症模型探究鲍姆纤孔菌(Sanghuangporus baumi)菌丝多糖对巨噬细胞RAW264.7的免疫调节作用

Immunomodulatory Effect of Polysaccharide Extracted from Sanghuangporus baumi Mycelia in LPS-Induced RAW264.7 Macrophages

鲍姆纤孔菌(Sanghuangporus baumi)是一种药用真菌,具有很高的活性,本文研究在脂多糖(LPS)诱导下鲍姆纤孔菌菌丝多糖对巨噬细胞RAW264.7的免疫调节作用。以鲍姆纤孔菌菌丝多糖(SJ)为研究对象,利用脂多糖(LPS)诱导的巨噬细胞RAW264.7建立炎症模型,实验分为空白对照组,LPS模型组,SJ处理组(SJ+LPS),采用Cell Counting Kit-8(CCK8)法检测SJ对巨噬细胞RAW264.7的增殖作用;利用酶联免疫(ELISA)法测定SJ对巨噬细胞RAW264.7一氧化氮(NO)、白介素-6(IL-6)、白介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)细胞因子分泌量的影响。此外,采用免疫印迹(Western-Blot)技术,研究SJ对RAW 264.7细胞生成IL-6、TNF-α、IL-1β和iNOS蛋白能力的影响。结果表明,不同浓度的SJ处理后巨噬细胞的活力与对照组相比分别增加了20.5%~50%;ELISA法结果表明:200μg/mL组的SJ极显著(P<0.001)抑制在炎症模型下巨噬细胞RAW264.7产NO的能力,比LPS模型组巨噬细胞NO、TNF-α、IL-6和IL-β分泌量分别降低了21.62%、29.99%、35.5%和50.03%。SJ呈剂量依赖的方式极显著(P<0.001)抑制由LPS诱导的促炎介质IL-6、TNF-α、IL-1β和iNOS表达,可减轻LPS引发的炎症反应,且与ELISA法检测到的细胞因子的分泌量变化结果相一致。综上,鲍姆纤孔菌多糖具有良好的免疫调节活性。

Sanghuangporus baumi is a kind of medicinal fungus with high activity.The aim of the present study was to investigate the immunoregulatory effects of Sanghuangporus baumi mycelia polysaccharides(SJ)on lipopolysaccharide(LPS)-induced RAW264.7 cells.In this study,Sanghuangporus baumi mycelia polysaccharides(SJ)were taken as the study subjects,and LPS-stimulated RAW264.7 macrophages were used as the cell model.The experiment was divided into three groups:Control group,LPS(model)and LPS+SJ groups.A CCK-8 kit was used to detect the viability of RAW264.7 cells.The effects of SJ on nitric oxide,and cytokines(IL-6,IL-1β,and TNF-α)production in LPS-stimulated RAW264.7 macrophages were measured by enzyme-linked immunoassay assay(ELISA).Furthermore,the effects of SJ on the protein levels of IL-6,TNF-α,IL-1β and inducible nitric oxide synthase(iNOS)were investigated by Western-Blot analysis.The results indicated that SJ increased 20.5%~50% cell viability at different concentrations compared to the control group.SJ at 200μg/mL potently inhibited the release of nitric oxide and cytokine production in LPS-stimulated RAW264.7(P<0.001),and compared with the LPS group,the secretion of NO,TNF-α,IL-6 and IL-1β decreased by 21.62%,29.99%,35.5% and 50.03%,respectively.Moreover,SJ also reduced LPS-induced protein expression levels of IL-6,TNF-α,IL-1β and iNOS in a dose-dependent fashion(P<0.001),and alleviated the inflammatory damage induced by LPS.And it was consistent with the change of cytokine secretion detected by ELISA method.The results of the present study suggest that Sanghuangporus baumi mycelia polysaccharides(SJ)have good immunomodulatory.