为研究硫酸化银耳多糖(sTPS70c)和硫酸化党参多糖(sCPPS50c)增强免疫作用机理,本试验以未修饰银耳多糖(TPStp)为对照,采用噻唑蓝(MTT)法和实时荧光定量PCR法测定了sTPS70c和sCPPS50c对鸡T淋巴细胞增殖及白细胞介素-2(IL-2)mRNA表达水平的影响.结果表明,多糖单独加入到外周血淋巴细胞时,sTPS70c和sCPPS50c几乎所有浓度均可显著刺激T淋巴细胞的增殖(P<0.05),而TPStp仅在浓度为3.125 μg/mL时显著刺激T淋巴细胞增殖(P<0.05);多糖与植物血凝素P(PHA-P)同时加入到外周血淋巴细胞时,sCPPS50c在浓度为0.391~1.563 μg/mL时显著刺激T淋巴细胞增殖(P<0.05);sTPS70c和sCPPS50c在浓度为1.563 μg/mL时可提高T淋巴细胞IL-2 mRNA的表达水平(P<0.05),其中sTPS70c对IL-2 mRNA的促表达作用显著强于TPStp(P<0.05),且在浓度为1.563 μg/mL时的作用最强.结果提示,硫酸化修饰可以提高多糖的淋巴细胞增殖活性及显著增强IL-2 mRNA的表达,且以sTPS70c的作用较强,这与取代度有一定的相关性.%In order to study the mechanism of sulfated polysaccharidses from Tremella (sTPS70c) and Condonpsis pilosula (sCPPS50c) on immunological enhancement activity, the effects of sTPS70c and sCPPS50c on T lymphocyte proliferation and the mRNA expression level of interleukin-2 (IL-2) were determined by methyl thiazolyl tetrazolium (MTT) assay and real-time fluorescent quantitative PCR assay while taking the unmodified polysaccharidses from Tremella (TPStp) as control.The results showed that in single adding into peripheral lymphocyte, sTPS70c and sCPPS50c almost at all concentrations and TPStp only at 3.125 μg/mL could significantly stimulate T lymphocyte proliferation (P<0.05).In simultaneous adding into peripheral lymphocyte with plant haemagglutinin P (PHA-P), sCPPS50c at 0.391 to 1.563 μg/mL significantly stimulated T lymphocyte proliferation (P<0.05).sTPS70c and sCPPS50c also could significantly promote mRNA expression level of IL-2 in T lymphocyte at 1.563 μg/mL (P<0.05), and the promoting effects of sTPS70c were significantly better than TPStp (P<0.05), especially at 1.563 μg/mL, the effect of sTPS70c was the strongest.These results indicate that sulfated modification can enhance the lymphocyte proliferation and mRNA expression of IL-2, and sTPS70c presents more stronger action, which is related to the degree of substitution at a centain extent.
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