【摘要】 目的:观察益元生血方提取物对免疫性血小板减少症(immune thrombocytopenia,ITP)患者T淋巴细胞亚群的影响。方法:(1)纳入ITP患者10例和健康志愿者10例,抽取受试者外周血制备淋巴细胞悬液,以10%、5%、2.5%、1.25%、0.625%浓度的益元生血方水提物干预ITP患者淋巴细胞悬液,培养时间分别为24 h、36 h、48 h、72 h,检测不同浓度、不同时间益元生血方水提物培养的ITP患者T淋巴细胞增殖的活性,以确定益元生血方水提物作用的最佳药物浓度和最佳培养时间。(2)根据试验需求,结合上述试验结果,设正常对照组、ITP未干预组和ITP干预组,正常对照组为健康志愿者淋巴细胞混悬液+完全RPMI-1640培养液,ITP未干预组为ITP患者淋巴细胞混悬液+完全RPMI-1640培养液,ITP干预组为ITP患者淋巴细胞混悬液+"最佳药物浓度"益元生血方灭菌提取物,培养时间为"最佳培养时间"。采用流式细胞术检测各组CD4~+、CD8~+、IFN-γ、IL-4、IL-17、CD4~+CD25~+T淋巴细胞亚群表达。结果:(1)当培养时间为36 h,益元生血方水提物药物浓度为5%时,T淋巴细胞增殖活性最佳。(2)与正常对照组比较,ITP未干预组CD4~+、CD4~+CD25~+、IL-4 T淋巴细胞表达及CD4~+/CD8~+比值均降低(P<0.05),CD8~+、IFN-γ、IL-17 T淋巴细胞表达及IL-17/CD4~+CD25~+、IFN-γ/IL-4比值均显著升高(P<0.05);与ITP未干预组比较,ITP干预组CD4~+、CD4~+CD25~+、IL-4 T淋巴细胞表达及CD4~+/CD8~+比值均升高(P<0.05),CD8~+、IFN-γ、IL-17 T淋巴细胞表达及IL-17/CD4~+CD25~+、IFN-γ/IL-4比值均显著降低(P<0.05)。结论:益元生血方可通过调节ITP患者T淋巴细胞亚群分化调节免疫,从而发挥治疗作用。更多还原

【Abstract】 Objective: To observe the influence of "Yiyuan Shengxue Prescription"extract on T lymphocyte subsets in patients with immune thrombocytopenia(ITP). Methods:(1)Ten patients with ITP and ten healthy volunteers were included.The peripheral blood of subjects was collected to prepare the lymphocyte suspension. The lymphocyte suspension was intervened by water extract of"Yiyuan Shengxue Prescription"with 10%,5%,2. 5%,1. 25% or 0. 625% concentration for 24 h,36 h,48 h or 72 h. The T lymphocyte proliferation was detected to explore the best drug concentration and culture time of water extract of"Yiyuan Shengxue Prescription".(2)Based on the experiment requirement and above experimental results,the normal control group(the lymphocyte suspension of healthy volunteers + RPMI-1640 culture medium),the ITP non-intervention group(the lymphocyte suspension of ITP patients + RPMI-1640 culture medium) and the ITP intervention group(the lymphocyte suspension of ITP patients + sterilized extract of "Yiyuan Shengxue Prescription"at the best drug concentration for the best culture time) were divided and treated. The T lymphocyte subsets expressions of CD4~+,CD8~+,IFN-γ,IL-4,IL-17 and CD4~+CD25~+were detected by flow cytometry. Results:(1)When the culture time was 36 h and the drug concentration of water extract of"Yiyuan Shengxue Prescription"was 5%,the proliferation activity of T lymphocyte was the best.(2)Compared with the normal control group,the expressions of CD4~+,CD4~+CD25~+,IL-4 in T lymphocytes and the ratio of CD4~+/CD8~+were all decreased(P < 0. 05) and the expressions of CD8~+,IFN-γ,IL-17 in T lymphocytes and the ratios of IL-17/CD4~+CD25~+and IFN-γ/IL-4 were all significantly increased(P < 0. 05) in the ITP non-intervention group. Compared with the ITP nonintervention group,the expressions of CD4~+,CD4~+CD25~+,IL-4 in T lymphocytes and the ratio of CD4~+/CD8~+were all increased(P < 0. 05) and the expressions of CD8~+,IFN-γ,IL-17 in T lymphocytes and the ratios of IL-17/CD4~+CD25~+and IFN-γ/IL-4 were all significantly decreased(P < 0. 05) in the ITP intervention group. Conclusion: "Yiyuan Shengxue Prescription"can regulate the differentiation of T lymphocyte subsets in patients with ITP to regulate the immune and further play the therapeutic role.更多还原