玉屏风散总多糖对小鼠派氏结淋巴细胞活化及功能影响的研究

The Effects of Yupingfeng Polysaccharides on Lymphocyte Activation and Function in Mice with Peyer's patches

通过研究玉屏风散总多糖对体外培养小鼠派氏结(Peyer's patches,PPs)淋巴细胞活化和功能变化的影响,探讨其对小鼠肠黏膜免疫应答的作用机制。体外培养小鼠PPs淋巴细胞,MTT法检测PPs淋巴细胞加入Con A协同不同浓度玉屏风散总多糖(0. 5、1、2、4、8 mg/m L)在作用不同时间(0、8、12、16、24、36 h)后的增殖变化; ELISA试剂盒检测细胞因子IL-2、IL-4、IFN-γ的含量变化;流式细胞术研究PPs淋巴细胞中T、B淋巴细胞的比例变化,进行T淋巴细胞的亚群分析。结果发现,不同浓度的玉屏风散总多糖均能增强PPs淋巴细胞的活性,单独作用时,浓度达到1 mg/m L以上,差异极显著(P <0. 01);与Con A刺激组比较,Con A与玉屏风散总多糖协同作用时,浓度达到2 mg/m L以上差异极显著(P <0. 01),且对PPs淋巴细胞增殖活化有明显促进作用(P <0. 01)。同时,玉屏风散总多糖可明显促进小鼠PPs淋巴细胞产生和分泌IL-2、IL-4、IFN-γ,改变PPs淋巴细胞中T、B淋巴细胞的比值,且可显著提高CD4+/CD8+的比值。结果表明,玉屏风散总多糖能显著提高小鼠PPs淋巴细胞活性,增强肠黏膜免疫功能。

To study the effect of Yupingfeng polysaceharide on the activation and function of lymphocytes in mouse Peyer＇s patches （PPs） cultured in vitro, the mechanism of the immune response to mouse intestinal mucosa was explored. Mouse PPs lymphocytes were cultivated in vitro, and MTY assay were performed on PPs lymphocytes treated with ConA plus different concentrations of Yupingfeng polysaccharide （0.5, 1,2,4, and 8 mg/mL） at different times （0,8,12,16 ,and 24 h） Proliferation changes were anlyzed after 36 h）. IL-2, IL-4 and IFN-γlevels were measured by ELISA kits. Flow cytometry was performed to study changes in the proportion of T and B lymphocytes in lymphocytes from PPs. The results showed that different concentrations of Yupingfeng polysaccharides enhanced the activity of PPs lymphocytes, and when Yuping feng polysaceharides was added alone, the concentration of 1 mg/ mL or more, the difference was extremely significant （ P 〈 0.01 ） ; compared with ConA alone stimulation group ConA and Yuping Feng polysaecharides acted synergistically. The difference was extremely significant （ P 〈 0.01 ） when the concentration reached 3 mg/mL, and had a significant effect on the proliferation and activation of PPs lymphocytes （P 〈0.01 ）. At the same time, Yupingfeng polysaecharide obviously promoted the production and secretion of IL-2, tL-4 and IFN-γ by mouse PPs lymphocytes, and the ratio of T and B lymphocytes in PPs lymphocytes was changed, having a significant increase of the CD4 ＋/CD8 ＋ ratio. The results showed that YPF-P significantly increases the activity of mouse PPs lymphoeytes and enhance intestinal mucosal immune function.