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本研究观察不同浓度马钱子碱对再生障碍性贫血(aplastic anenia,AA)患者T淋巴细胞分泌TNF-α、IFN-γ、IL-4的水平及其增殖的影响,以探讨马钱子治疗AA的作用机制。AA患者及健康志愿者各10例,分离外周血T淋巴细胞并对其进行培养和纯化。将AA患者T细胞分为马钱子碱0、100、200和400μg/ml 4个干预组;健康志愿者T细胞为正常对照组,不作干预。培养72小时后,用ELISA法检测各组细胞培养上清液中TNF-α、IFN-γ、IL-4的水平,MTT比色法检测AA各组T细胞的增殖状况。结果表明,AA患者T细胞培养上清液中TNF-α、IFN-γ水平较正常对照组均明显升高,而IL-4水平较正常对照组明显降低;AA患者T细胞经马钱子碱作用后,TNF-α、IFN-γ水平均明显降低,且呈剂量依赖关系,而作用前后IL-4水平无明显变化。马钱子碱100、200和400μg/ml浓度对AA患者T细胞生长抑制率分别为(13.61±4.31)%、(14.28±4.31)%、(15.12±4.56)%,3个组间的比较无明显差异。结论:马钱子碱通过降低AA患者TNF-α、IFN-γ造血负调控因子的水平和抑制T细胞增殖而发挥治疗作用。本研究为马钱子治疗AA提供了实验依据。
This study was to investigate the effects of different concentrations of brucine on TNF-α, IFN-γ and IL-4 secreted by T lymphocytes in patients with aplastic anemia (AA) AA mechanism of action. AA patients and healthy volunteers in each of 10 cases, peripheral blood T lymphocytes were isolated and cultured and purified. AA patients T cells were divided into brucine 0,100,200 and 400μg / ml intervention group; healthy volunteers T cells as normal control group, without intervention. After cultured for 72 hours, the levels of TNF-α, IFN-γ and IL-4 in the cell culture supernatants of each group were detected by ELISA. The proliferation of T cells in AA groups was detected by MTT colorimetry. The results showed that the levels of TNF-α and IFN-γ in T cell culture supernatant of patients with AA were significantly higher than those in normal control group, while the levels of IL-4 were significantly lower in AA patients than in normal control group. T cells in AA patients were treated with brucine After treatment, the levels of TNF-αand IFN-γwere significantly reduced and in a dose-dependent manner, while the levels of IL-4 had no significant changes before and after treatment. The inhibitory rates of brucine, 100, 200 and 400 μg / ml strychnine on patients with AA were (13.61 ± 4.31)% and (14.28 ± 4.31)%, (15.12 ± 4.56)%, respectively Significant differences. Conclusion: Brucine plays a therapeutic role in reducing the level of TNF-α and IFN-γ negative regulatory factor in AA and inhibiting T cell proliferation. This study provided the experimental evidence for the treatment of AA by Strychnos.