三氧化二砷对MRL/lpr狼疮鼠淋巴细胞亚群和细胞因子表达的影响

Effects of Arsenic Trioxide on Expression of Cytokines and Lymphocyte Subsets in MRL/lpr Mice

目的探讨三氧化二砷(arsenic trioxide,ATO)对MRL/lpr狼疮鼠淋巴细胞亚群和细胞因子表达的影响。方法MRL/lpr狼疮鼠随机分为ATO组(0.4mg·kg-1·d-1,ip)、环磷酰胺(CTX)组(50mg·kg-1每周1次,ip)和生理盐水(NS)组(按体重取量,ip),给药2个月。取脾脏制成脾细胞悬液,加入佛波醇酯(PMA)和ionomycin活化,同时加monensin抑制蛋白转运,于37℃,5%CO2培养4h后,用四色流式细胞术测脾脏CD3+(T)细胞和CD3+CD4+(Th)细胞的百分率以及CD3+CD4+(Th)细胞内细胞因子IFN-γ、IL-4、IL-10和IL-12的水平;分离血清,用ELISA法测血清中抗ds-DNA抗体、IFN-γ、IL-4、IL-10和IL-12的浓度。结果①给药后,ATO组小鼠血清的抗ds-DNA抗体水平较给药前明显下降(P<0.01),而NS对照组则明显升高(P<0.05)。②ATO组CD3+细胞和CD3+CD4+百分率均显著低于NS组(P<0.01);③ATO组的血清IFN-γ和IL-12浓度均低于NS组(前者P<0.01,后者P=0.018);④ATO组Th细胞内IFN-γ、IL-4、IL-10和IL-12表达均显著低于NS组(P<0.01)。结论三氧化二砷能显著降低MRL/lpr狼疮鼠血清抗ds-DNA抗体的水平,抑制T细胞和Th细胞增生和活化功能,降低IFN-γ和IL-12的血清水平和Th细胞内IFN-γ、IL-4、IL-10和IL-12的水平。

OBJECTIVE To investigate the effects of arsenic trioxide （ATO） on expression of cytokines and lymphocyte subsets proliferation in splenic ceils of MRL/lpr mice. METHODS MRL/lpr mice were divided into 3 different groups. The 3 groups received arsenic trioxide （ATO, 0.4 mg-kg^-1·d^-1）, cyclophosphamide （CTX, 50 mg·kg^-1qw^-1） and sodium chloride respectively. The treatment was lasted for 2 months. The contents of the CD3^＋（T）cells, CD3^＋CD4^＋（Th）cells and the IL-4, IL-10 and IL-12 were detected using single-cell measurement of intracellular cytokines by flow cytometry after polyclonal stimulation with PMA and ionomycin for 4 h in 5% CO2. Serum levels of IFN-y, and IL-4 were assayed using the Mouse cytokines ELISA Kit. RESULTS ① The production of ds-DNA antibody was significantly decreased in ATO group （P〈 0.01 ）, while it was dramatically increased in the NS groups（P〈0.05 ） after the treatment. ②Compared with NS group, CD^3＋cells and CD3^＋CD4^＋cells in ATO group were decreased significantly （P〈0.01）. ③The serum levels of IFN-y and IL-12 were significantly reduced in ATO group （P〈0.01 and P=0.018 ）. ④The intracellular levels of IFN-y, IL-4, L-10 and IL-12 produced by CD3^＋CD4^＋（Th）cells in ATO group were significantly lower than those in NS group （P〈0.01）. CONCLUSION Arsenic trioxide can reduce the production ofds-DNA antibody, inhibit the activation and proliferation of both T cells and Th subsets in MRL/lpr mice, and hence decrease the serum levels of IFN-y, IL-12 and the levels of IFN-y, IL-4, IL-10 and IL-12 yielded by Th cells.