黄芪多糖对鸡淋巴细胞体外增殖及脾脏淋巴细胞分泌细胞因子和相关mRNA表达的影响

Effect of Astragalus Polysacchride on Cell Proliferation of Lymphocytes and Splenic Lymphokines Secretion and Related mRNA Expression in Spleen of Chick

【目的】探讨黄芪多糖(APS)对雏鸡不同免疫组织淋巴细胞体外增殖和脾脏淋巴因子分泌及相关基因mRNA表达的影响。【方法】将浓度为20、40、80、160以及320μg/mL的APS分别加入用刀豆蛋白(ConA)刺激的体外培养雏鸡外周血、胸腺、脾脏和法氏囊淋巴细胞中,采用MTT比色法测定淋巴细胞增殖情况,ELISA检测脾脏淋巴细胞中Th1型细胞因子(IL-2、TNF-β、IFN-γ)和Th2型细胞因子(IL-4、IL-6)的含量,RT-qPCR检测细胞因子相关基因以及核转录因子TBX21和Gata-3基因mRNA表达量。【结果】浓度为80～320μg/mL的APS可显著刺激外周血、胸腺、脾脏和法氏囊淋巴细胞增殖,明显促进细胞因子IL-2、TNF-β、IFN-γ、IL-4、IL-6的分泌以及相关基因mRNA的表达,并可显著提高TBX21和Gata-3的mRNA表达量,其中APS为160μg/mL效果最佳。【结论】APS可通过促进淋巴细胞Th1和Th2细胞因子分泌,提高淋巴细胞增殖,从而参与调解机体细胞免疫。

[Objective] The aim of this study was to investigate the effect of Astragalus polysaccharide LAPS） on the cell proliferation of lymphocytes from different tissues, and secretion of spleen cytokines and related gene mRNA expression in chicks. [Method] APS of different concentrations （20,40,80,160 and 320 μg/mL） were added into cultured lymphocytes isolated from peripheral blood, thymus, spleen and bursa, respectively. All those tissues were pretreated with ConA. MTT and enzyme linked immunosorbent assa LELISA） were used to detect the lymphocyte proliferation and the contents of Thl and Th2 type cytokines in spleen lymphocytes, respectively. The gene expressions oflL-2, TNF-β, IFN-T, 1L-4, IL-6 and transcription factors TBX21 and Gata-3 in spleen lymphocytes were determined by RT-qPCR. [Result] The results showed that APS supplementation ranged from 80 to 320 μg/mL significantly induced the proliferation of lymphocytes isolated from immunologic tissues, and promoted the secretions of IL-2, TNF-β, IFN-γ, IL-4, IL-6 and the mRNA expressions of related genes. Meanwhile, APS supplementation remarkably improved the mRNA abundance of TBX21 and Gata-3. The concentration of APS at 160 μg/mL showed the best effect in promoting lymphocytic proliferation. [Conclusion] The results suggested that APS could strengthen cellular immune functions by promoting the secretions of Thl and Th2 cytokines in spleen and increasing the proliferation of lymphocytes in chicks.