目的:探讨雪灵芝水溶性提取物对淋巴细胞增殖及其Th1型细胞因子表达的影响。方法:体外培养的小鼠脾淋巴细胞及人外周血单个核细胞(PBMC)经不同浓度雪灵芝水提物处理。采用MTT法检测小鼠脾淋巴细胞增殖水平;应用ELISA双抗体夹心法检测小鼠脾淋巴细胞培养上清中IL-2和IFN-γ水平;RT-PCR与实时定量PCR检测人PBMC中NF-кB p65、IL-2及IFN-γ的mRNA表达。结果:雪灵芝水提物在0.025~0.2 mg/ml浓度范围,对小鼠脾淋巴细胞增殖具有浓度依赖性的促进作用;0.05 mg/ml和0.1 mg/ml雪灵芝处理组IL-2水平高于对照组(P〈0.05),0.4 mg/ml雪灵芝处理组IFN-γ水平高于对照组(P〈0.05);相对于对照组,0.2 mg/ml雪灵芝处理组人PBMC的NF-кB p65、IL-2及IFN-γmRNA相对表达系数分别为204.76、4.31和40.46。结论:雪灵芝水提物对小鼠脾淋巴细胞增殖以及小鼠脾淋巴细胞、人PBMC中Th1型细胞因子的表达具有促进作用。
Objective:To investigate the effects of Arenaria kansuensis aqueous extract(AKAE) on lymphocytes proliferation and Th1 cytokine expression.Methods:Mice spleen lymphocytes and human peripheral blood monouclear cells(PBMC) were treated with AKAE at various concentration in vitro;the proliferation level of mice spleen lymphocytes was observed by MTT assay;the level of IL-2 and IFN-γ produced by mice spleen lymphocytes were tested by enzyme-linked immunosorbent assay(ELISA);RT-PCR and realtime PCR were used to detect the mRNA expression of NF-кB p65,IL-2 and IFN-γ in human PBMC.Results:The proliferation of mice spleen lymphocytes was dose-dependently enhanceed by AKAE treatment at concentration of 0.025 ~ 0.2 mg/ml;in culture supernatant of mice spleen lymphocytes,the level of IL-2 was increased after AKAE treatment at dose of 0.05 mg/ml and 0.1 mg/ml(P0.05),and the level of IFN-γ was increased after AKAE treatment at dose of 0.4 mg/ml(P0.05);compared with control group,the relative expression of NF-кB p65,IL-2 and IFN-γ mRNA in human PBMC treated by 0.2 mg/ml AKAE were 204.76,4.31 and 40.46 respectively.Conclusion:AKAE could enhance the proliferation of mice spleen lymphocytes and promote the Th1 cytokine expression of mice spleen lymphocytes and homo PBMC.