【摘要】 目的比较体外给药时灵芝孢子多糖(Gl-SP)和灵芝破壁孢子多糖(Gl-BSP)对小鼠脾淋巴细胞及腹腔巨噬细胞免疫调节活性的影响。方法噻唑蓝(MTT)法检测淋巴细胞增殖、混合淋巴细胞培养反应(MLR)、自然杀伤细胞(NK细胞)活性;比色分析检测巨噬细胞吞噬中性红的能力;流式细胞术测定脾T细胞亚群;ELISA法测定白细胞介素-2(IL-2)、γ干扰素(IFN-γ)含量,生物法测定肿瘤坏死因子α(TNF-α)活性,Griess法测定NO含量。结果当质量浓度为0.2～12.8mg/L时两种多糖均可促进淋巴细胞增殖及丝裂原诱导的淋巴细胞增殖,提高NK细胞活性,增强腹腔巨噬细胞吞噬能力并促进TNF-α和NO分泌。两种多糖均可促进MLR,在质量浓度为12.8mg/L时,Gl-BSP促增殖作用明显强于Gl-SP,Gl-SP或Gl-BSP(0.2～12.8mg/L)可显著增加IL-2、IFN-γ的分泌,但相同浓度的Gl-BSP增加IL-2和IFN-γ分泌的作用均强于Gl-SP。在一定的浓度范围内,两种多糖均可显著增加双向MLR反应中CD3+T、CD4+T和CD8+T细胞百分率,Gl-BSP在质量浓度为0.2～12.8mg/L或3.2～12.8mg/L时,增加CD4+或CD8+T细胞的作用较Gl-SP更为显著。在质量浓度为0.2～0.8mg/L时,Gl-BSP组的CD4+T和CD8+T细胞的比值大于Gl-SP组。结论Gl-SP和Gl-BSP在体外试验中具有相似的免疫调节活性,Gl-BSP的某些免疫调节作用强于Gl-SP。更多还原

【Abstract】 Objective: To compare the immunomodulatory effects of spore polysaccharides (Gl-SP) and broken spore polysaccharides (Gl-BSP) isolated from Ganoderma lucidum(Leyss et Fr.) Karst. on murine splenic lymphocytes and peritoneal macrophages in vitro. Methods: Mixed lymphocyte culture reaction (MLR), lymphocyte proliferation in the presence or absence of mitogen, and the cytotoxic activity of splenic natural killer (NK) cells were detected with MTT assay in vitro. The percentage of phagocytosis of neutral red (NR) by mouse peritoneal macrophages was detected by colorimetric assay. Splenic T-lymphocyte subpopulations were measured with flow cytometry(FCM). IL-2, IFN-γ and TNF-α in the culture supernatants were detected by ELISA and biological assay. Nitric oxide (NO) production was examined by Griess reaction. Results: At the concentration range of 0.2-12.8 mg/L, Gl-SP and Gl-BSP were shown to increase lymphocyte proliferation in the presence or absence of mitogen, enhance NK cytotoxic activity, augment the production of TNF-α and NO in Gl-SP-or Gl-BSP-activated macrophages, as well the percentage of phagocytosis of NR by macrophages in vitro. Both Gl-SP and Gl-BSP could promote MLR, however, at the dose of 12.8mg/L, Gl-BSP showed higher activity than Gl-SP in the proliferation of lymphocytes. These two kinds of polysaccharide could significantly increase the secretion of IL-2 and IFN-γ in double-way MLR at the concentrations of 0.2-12.8 mg/L, but Gl-BSP had stronger effects than Gl-SP at the same concentrations. Both Gl-SP and Gl-BSP could increase the ratio of T-lymphocyte subpopulations in double-way MLR. At the concentrations of 0.2-12.8 mg/L or 3.2-12.8 mg/L, Gl-BSP demonstrated more significant activity in increasing the percentage of the CD4+ or CD8+ subset than Gl-SP. At the concentrations of 0.2-0.8 mg/L, the ratio of the CD4+ and CD8+ subset in the Gl-BSP treated group was higher than that of the Gl-SP treated group. Conclusion:Gl-SP and Gl-BSP have sim-ilar immunomodulatory effects in vitro, as though the immunomodulatory effects of Gl-BSP are stronger than that of Gl-SP.更多还原