基因芯片研究云芝糖肽对健康成人外周血单个核细胞Toll样蛋白受体信号转导通路的影响

Regulatory effects of polysaccharopeptide on Toll-like receptor signaling pathway in human PBMC by cDNA microarray

目的利用基因芯片技术检测PSP对健康成人外周血单个核细胞(PBMC)Toll样蛋白信号转导通路的影响,探索云芝糖肽的免疫调节作用机制。方法体外分离、培养健康成人外周血单个核细胞,摸索云芝糖肽的最佳作用浓度,再分为对照组、云芝糖肽组,72h后收获细胞抽提总RNA,应用Toll样蛋白受体信号转导基因芯片检测PSP对健康成人外周血单个核细胞TLRs信号传导通路中相关基因的变化。结果12.5μg/mL的云芝糖肽可显著促进健康成人PBMC增殖转化(P<0.01);经云芝糖肽作用后上调≥2倍的基因有22个,占18.8%(如γ-干扰素、干扰素-γ-诱导蛋白-10等),下调≥2倍的基因有23项,占19.7%(如Toll作用蛋白、共刺激分子CD86等)。结论云芝糖肽可以调节免疫细胞Toll样蛋白受体转导途径中相关基因的表达,为进一步探讨云芝糖肽的免疫调节作用机理提供了依据。

Objective To detect the effects of polysaccharopeptide （PSP） on Toll-like receptor signaling pathway in humar, PBMC by cDNA microarray technique, so as to explore the immunoregulatory mechanism of PSP. Methods The human PBMC were isolated and cultured in vitro, and then divided into control group and PSP group. The total RNA was collected after 72 hours and cDNA microarray （Oligo GEArray Human Toll-like receptor signaling pathway microarray） was used to screen the genes which showed significant changes in expression of Toll-like receptor signaling pathway in human PBMC induced by PSP. Results PSP could significantly promote human PBMC proliferation when the concentration was 12.5 μg/mL compared with control group （ P 〈0.01）. The microarray analysis showed that 22 genes （ 18.8% ）, such as IFN-γ and CXCL10, were up-regulated over two-times by PSP and 23 genes （ 19.7% ）, such as TOLLIP, CD86, were down-regulated over two-times by PSP. Conclusion PSP can regulate the related gene expression of Toll-like receptor signaling pathway in human PBMC, and supply evidence for further exploring the immunoregulatory mechanism of PSP.