目的:观察三氧化二砷(ATO)对MRL/lpr狼疮小鼠脾脏CD4+T细胞分泌IFN-γ及其mRNA的影响。方法:免疫磁珠分别分选18～20周MRL/lpr狼疮小鼠和C57BL/6J正常对照小鼠脾脏CD4+T细胞,采用流式细胞术鉴定细胞纯度。PHA-p(20μg/mL)和IL-2(1000IU/mL)常规刺激48h后进行如下实验:(1)不同浓度ATO(0.5、1.0和2.0μmol/L)作用24h;(2)1μmol/LATO作用不同时间(12、24和48h);(3)不同药物处理组:①PBS组:空白对照;②ATO组:1μmol/LATO;③5-氮杂胞苷(5-AzaC)组:1μmol/L5-AzaC;④ATO+5-AzaC组:1μmol/LATO+1μmol/L5-AzaC。酶联免疫吸附法(ELISA)测定培养上清液IFN-γ的表达量;实时荧光定量PCR法检测不同药物处理组中IFN-γmRNA的表达情况。结果:①1.0μmol/LATO作用24h对细胞存活率无明显影响。②1mmol/lATO作用24h能抑制MRL/lpr小鼠脾脏CD4+T细胞IFN-γ的转录和翻译水平。③经5-AazC处理后MRL/lpr和C57BL/6J小鼠脾脏CD4+T细胞IFN-γ的转录和翻译水平升高,1mmol/LATO作用24h能抑制其异常活化状态。④MRL/lpr小鼠脾脏CD4+T细胞IFN-γ的分泌和表达水平均较C57BL/6J小鼠明显升高。结论:1mmol/LATO作用24h能在一定程度上有效抑制活化状态下MRL/lpr小鼠脾脏CD4+T细胞IFN-g的异常分泌,下调其mRNA表达水平而不影响其存活率。 Objective: To observe the effects of arsenic trioxide (ATO) on the secretion of IFN-γ and its mRNA from splenic CD4 + T cells of MRL / lpr lupus mice. Methods: Immunomagnetic beads were used to separate CD4 + T cells from MRL / lpr lupus mice and C57BL / 6J normal mice spleen from 18 to 20 weeks respectively. The cell purity was determined by flow cytometry. The following experiments were carried out 48h after conventional stimulation with PHA-p (20μg / mL) and IL-2 (1000IU / mL): (1) ATO at different concentrations (0.5,1.0 and 2.0μmol / L) (12,24 and 48h); (3) different drug treatment groups: ①PBS group: blank control; ②ATO group: 1μmol / LATO; ③5-azacytidine (5-AzaC) group: 1μmol / L5-AzaC ; ④ATO + 5-AzaC group: 1μmol / LATO + 1μmol / L5-AzaC. The expression of IFN-γ in culture supernatants was determined by enzyme-linked immunosorbent assay (ELISA). The expression of IFN-γ mRNA in different drug-treated groups was detected by real-time fluorescence quantitative PCR. Results: ① The effect of 1μmol / L ATO for 24 hours had no significant effect on the cell survival rate. ②1mmol / lATO 24h can inhibit the transcription and translation of IFN-γ in splenic CD4 + T cells of MRL / lpr mice. ③ The transcription and translation of IFN-γ in splenic CD4 + T cells of MRL / lpr and C57BL / 6J mice after 5-AazC treatment were increased, and the abnormal activation was inhibited by 1mmol / L ATO for 24h. ④ The secretion and expression of IFN-γ in CD4 + T cells of MRL / lpr mice were significantly higher than those in C57BL / 6J mice. CONCLUSION: 1 mmol / L ATO for 24 h can effectively inhibit the abnormal secretion of IFN-g from splenic CD4 + T cells in activated MRL / lpr mice to a certain extent and down-regulate the mRNA expression without affecting the survival rate.