防风多糖体内外免疫调节活性研究

Study on Evaluation of Immunoregulatory Activity of Polysaccharides from Fangfeng(Saposhnikovia divaricata)in vitro and in vivo

目的采用体内外多指标免疫调节活性评价方法,研究防风多糖体内外免疫功能的调节作用。方法体外实验选用RAW 264.7小鼠巨噬细胞系,采用CCK-8法检测巨噬细胞的增殖能力,中性红染色法检测巨噬细胞的吞噬能力,Griess法测定巨噬细胞NO释放量,酶联免疫吸附法(ELISA)测定巨噬细胞肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)和白细胞介素-6(IL-6)释放量。体内实验选用环磷酰胺诱导的免疫低下BALB/c小鼠,通过碳粒廓清实验探讨防风多糖对小鼠网状内皮系统吞噬功能,以小鼠免疫器官指数评价防风多糖对小鼠免疫器官的影响,并采用ELISA法测定小鼠体内免疫球蛋白(IgM、IgG、IgA)和补体(C3、C4)含量。结果体外实验结果表明,防风多糖能够显著提高巨噬细胞的增殖和吞噬能力,并促进巨噬细胞释放NO、TNF-α和IL-6,但对IL-1β的释放量影响不显著。体内实验结果表明,防风多糖可使免疫低下小鼠模型的吞噬能力和免疫器官指数显著提高,并使免疫低下小鼠体内的免疫球蛋白(IgM、IgG、IgA)和补体(C3、C4)的含量升高。结论防风多糖在体内外水平均具有较好的免疫调节活性,参与调节机体的特异性免疫和非特异性免疫。

Objective The immunoregulatory activity of polysaccharides from Fangfeng(Saposhnikovia divaricata)was evaluated in vitro and in vivo.Methods In vitro,RAW 264.7 mouse macrophage system was used to detect the proliferation of macrophages by CCK-8 method,the phagocytosis of macrophages by neutral red staining,the NO release of macrophages by Griess method,and the levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and interleukin-6(IL-6)of macrophages were measured by ELISA.In vivo experiments were carried out in immunocompromised BALB/c mice induced by cyclophosphamide.The phagocytosis function of polysaccharides from Fangfeng(Saposhnikovia divaricata)on reticuloendothelial system of mice was studied by carbon particle clearance test.The effect of polysaccharides from Fangfeng(Saposhnikovia divaricata)on immune organs of mice was evaluated by mouse immune organ index.The contents of immunoglobulin(IgM,IgG,IgA)and complement(C3,C4)in mice were determined by ELISA.Results The results in vitro showed that polysaccharides from Fangfeng(Saposhnikovia divaricata)could significantly improve the proliferation and phagocytosis of macrophages,and promote the release of NO,TNF-α and IL-6,but the effect on the release of IL-1β was not significant.The results of in vivo experiments showed that polysaccharide from Fangfeng(Saposhnikovia divaricata)could significantly improve the phagocytic capacity and immune organ index of immunosuppressed mice,and increase the contents of immunoglobulin(IgM,IgG,IgA)and complement(C3,C4)in immunosuppressed mice.Conclusion Polysaccharides from Fangfeng(Saposhnikovia divaricata)have good immunoregulatory activity both in vivo and in vitro,and participate in regulating specific and non-specific immunity of the body.