马钱子碱对再生障碍性贫血患者T细胞分泌功能和增殖能力的影响

Effect of Brucine on Secretion Function and Proliferation Capability of T Lymphocytes in Patients With Aplastic Anemia

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。

The aim of this study was to investigate the effect of brucine on secretion of TNF-α,IFN-γ,IL-4 and proliferation of T lymphcytes in patients with aplastic anemia（AA）,and to explore its mechanism.Peripheral blood T lymphocytes from 10 patients with AA and 10 healthy volunteers were isolated,purified and cultured.T lymphocytes from the patients were divided into 0,100,200 and 400 μg/ml brucine-treated groups.T lymphocytes from healthy volunteers were used as control group.After being cultured for 72 hours,the levels of TNF-α,IFN-γ,IL-4 in the supernatant of cultured T lymphocytes from AA patients were detected by ELISA,and the proliferation of T lymphocytes from AA patients was detected by MTT.The results showed that compared with the normal control group,the levels of TNF-α and IFN-γ in the culture supernatant significantly increased,and IL-4 was significantly decreased.The levels of TNF-α and IFN-γ in the culture supernatant of brucine treated groups were lower,and were dependent on the concentration of brucine.However,the levels of IL-4 were found to be not obviously changed.The inhibition rate of T lymphocytes in 100,200 and 400μg/ml brucine-treated groups were（13.61± 4.31）%,（14.28±4.31）% and（15.12±4.56）% respectively,among which the differences were not statistically significant.It is concluded that the brucine can reduce the levels of TNF-α and IFN-γ through inhibiting the proliferation of T lymphocytes in AA patients,which provides experimental basis for therapy of AA patients.