三氧化二砷对MRL/lpr小鼠CD4^+T细胞增殖和IFN-γ、IL-4分泌的调节作用  

Regulatory Effects of Arsenic Trioxide on CD4～+T Lymphocyte Proliferation and IFN-γ and IL-4 Expression in Splenocytes of MRL/lpr Mice

作　　者：夏晓茹[1] 邹贤斐[1] 张挺[1] 孙莉[1] 朱小春[1] 

机构地区：[1]温州医学院附属第一医院,浙江温州325000

出　　处：《中国药学杂志》2009年第11期836-841,共6页Chinese Pharmaceutical Journal

基　　金：浙江省自然科学基金资助项目(M303606);温州市科技局资助项目(Y2003A007)

摘　　要：目的探讨三氧化二砷(ATO)对MRL/lpr狼疮小鼠脾脏CD4+T细胞增殖以及IFN-γ和IL-4转录和翻译水平的影响。方法免疫磁珠分别分选15只MRL/lpr狼疮小鼠和15只C57BL正常对照小鼠脾脏CD4+T细胞,进而将CD4+T细胞随机分为空白组(RPMI1640),对照组(PHA20mg·L-1+IL-21000u·mL-1),ATO组[PHA20mg·L-1+IL-21000u·mL-1(48h)后+ATO1.0μmol·L-1],体外共培养至72h。采用细胞计数试剂盒(CCK-8)检测3组CD4+T细胞的增殖和抑制水平;酶联免疫吸附反应(ELISA)检测上清液中IFN-γ和IL-4水平;逆转录-聚合酶链反应(RT-PCR)检测上述细胞因子基因的转录水平。结果①分选后获得的CD4+T细胞纯度为(94.12±1.08)%,细胞存活率为(95.82±3.41)%;②MRL/lpr小鼠对照组CD4+T细胞的增殖水平(A值)(0.343±0.019)较空白组(0.054±0.008)高(P<0.001);却低于C57BL小鼠对照组的水平(0.375±0.018)(P<0.05);③MRL/lpr和C57BL小鼠ATO组CD4+T细胞增殖水平(A值)分别为(0.324±0.013)和(0.362±0.013),均低于对照组(P<0.05,P<0.05);④MRL/lpr小鼠对照组IFN-γ的表达水平[(562.287±60.345)ng·L-1]明显高于C57BL小鼠对照组[(253.208±59.434)ng·L-1](P<0.05);ATO对MRL/lpr小鼠IFN-γ的抑制程度大于C57BL小鼠(P<0.05);MRL/lpr小鼠对照组的IL-4表达水平[(41.332±8.039)ng·L-1]与ATO组[(23.046±3.253)ng·L-1]和C57BL小鼠对照组[(29.303±8.038)ng·L-1]比较,也具有明显的统计学差异(P<0.05和P<0.05);ATO对MRL/lpr小鼠IL-4的抑制程度高于C57BL小鼠(P<0.05);⑤在转录水平上,与C57BL小鼠相比,ATO对MRL/lpr小鼠IFN-γmRNA和IL-4mRNA的抑制程度更加明显(P<0.05和P<0.05);ATO对IFN-γ和IL-4表达水平的抑制和转录水平的抑制正相关,r=0.720(P<0.001)和r=0.557(P<0.01)。结论MRL/lpr小鼠存在CD4+T细胞的异常增殖,ATO能在不影响细胞存活率的浓度下抑制MRL/lpr小鼠CD4+T细胞的异常增殖活化,通过抑制其细胞因子IFN-γ和IL-4的mRNA分泌影响两者的表达,且对MRL/lpr小鼠抑制程度大于C57BL小鼠,表明ATO能抑制狼OBJECTIVE To evaluate the effects of arsenic trioxide （ATO） on the proliferation ofCD4^＋T lymphocytes and the transcription and translation of interferon-γ（IFN-γ） and interleukin-4 （IL-4） in MRL/lpr and C57BL mice. METHODS CD4^＋T cells in spleencytes from 15 MRL/lpr and C57BL mice were separated by immunomagnetic beads respectively, and then divided randomly into three groups： blank group（RPMI1640）, control group（PHA 20 mg·L^-1＋IL-2 1 000 u-mL^-1）, ATO group[PHA 20 mg·L^-1＋ IL-2 1 000 u-mL^-1 （48 h）after＋ATO 1.0 μmol·L^-1], cultured for 72 h. The proliferation and suppression level in three groups of CD4^＋T cells were evaluated by cell counting kit-8（CCK-8）. The transcription and translation levels of IFN-y and IL-4 were measured using RT-PCR and ELISA respectively. RESULTS （1）The purity of CD4^＋T cell was （94.12±1.08）% and the survival rate was （95.82＋3.41） % after separation. （2）The proliferation level of CD4^＋T cell in control group （A value） （0.343± 0.019） was higher than that in blank group （0.054±0.008） in MRL/lpr mice, P〈0.001; but lower than that in control group in C57BL mice （P〈0.05）. （3）The proliferation levels of CD4^＋T cell in ATO groups in MRL/lpr and C57BL mice were （0.324±0.013） and （0.362±0.013） respectively, both of which were lower than that in control group （P〈0.05, P〈0.05）. （4）The IFN-γ expression level in control group in MRL/lpr mice was much higher than that in C57BL mice, which were [（562.287±60.345） ng·L^-1] and [（253.208±59.434） ng·L^-1] respectively, （P〈0.05） . ATO suppression effect on IFN-y in MRL/lpr mice was much higher than that in C57BL mice （P〈0.05）Comparing the IL-4 expression in control group [（41.332 ± 8.039） ng·L^-1 with ATO group[（23.046 ±3.253） ng·L^-1] in MRL/Ipr mice and control group[（29.303 ± 8.038） ng·L^-1] in C57BL mice, it showed significant difference （P〈0.05 and P〈0.05）. ATO suppression effects

关 键 词：系统性红斑狼疮 三氧化二砷 CD4^+T细胞 γ-干扰素 白介素-4 

分 类 号：R593.241[医药卫生—内科学]