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目的:探讨砒霜(Arsen ic Trioxide,As2O3)对体外MRL/lpr狼疮小鼠脾Th细胞及分泌相关细胞因子IFN-γ和IL-10的调节作用。方法:通过免疫磁珠分别分选MRL/lpr和C57BL小鼠脾Th细胞,Th细胞随机分PHA组和空白组,采用酶联免疫吸附反应(ELISA)和逆转录-聚合酶链反应(RT-PCR)法比较MRL/lpr和C57BL小鼠Th细胞分泌IFN-γ和IL-10及其mRNA水平的差异,然后观察As2O3对MRL/lpr小鼠Th细胞分泌IFN-γ和IL-10及其mRNA水平的影响。细胞计数试剂盒(CCK-8)观察不同浓度(0.5、1.0、1.5、2.0μmol/L)及作用时间As2O3对Th细胞增殖的影响。结果:(1)与C57BL相比,MRL/lpr小鼠各组Th细胞分泌的IFN-γ和IL-10水平显著增高,1.0μmol/L的As2O3能明显抑制MRL/lpr小鼠各组上述因子的分泌;(2)PHA组的IL-10 mRNA和IFN-γmRNA水平在MRL/lpr小鼠中表达均高于C57BL小鼠,As2O3能抑制MRL/lpr小鼠Th细胞空白组和PHA组中IL-10mRNA和IFN-γmRNA的高表达;(3)除0.5μmol/L的As2O3能刺激Th细胞的增殖,≥1.0μmol/L的As2O3能抑制Th细胞的生长,但浓度增加对细胞存活率有明显的影响。结论:适当浓度的砒霜能有效下调Th细胞因子IL-10和IFN-g mRNA的异常分泌及而影响因子的表达,但并不产生过度的免疫抑制,并能下调MRL/lpr狼疮小鼠脾脏Th细胞的异常增殖而不影响其存活率,从而调节狼疮鼠的异常免疫功能,可能是砒霜治疗SLE的重要机制之一。
AIM: To investigate the regulatory effect of Arsen ic Trioxide (As2O3) on spleen Th cells and secretion-related cytokines IFN-γ and IL-10 in murine MRL / lpr lupus mice. Methods: The splenic Th cells of MRL / lpr and C57BL mice were respectively sorted by immunomagnetic beads. Th cells were randomly divided into PHA group and blank group. ELISA and reverse transcription polymerase chain reaction (RT- PCR) was used to compare the secretion of IFN-γand IL-10 and its mRNA level in Th cells of MRL / lpr and C57BL mice. Then the effect of As2O3 on the secretion of IFN-γ and IL-10 and mRNA of Th cells in MRL / lpr mice Horizontal impact. The cell counting kit (CCK-8) was used to observe the effect of different concentrations (0.5,1.0,1.5,2.0μmol / L) and action time As2O3 on the proliferation of Th cells. Results: (1) Compared with C57BL, the levels of IFN-γ and IL-10 secreted by Th cells in MRL / lpr mice were significantly increased. 1.0μmol / L As2O3 significantly inhibited the above factors in MRL / lpr mice (2) The levels of IL-10 mRNA and IFN-γmRNA in PHA group were higher than those in C57BL mice in MRL / lpr mice. As2O3 inhibited the IL-10 mRNA and IFN- (3) Except that 0.5μmol / L As2O3 could stimulate the proliferation of Th cells, ≥1.0μmol / L As2O3 could inhibit the growth of Th cells, but the cell survival rate was obviously increased with the increase of concentration Impact. CONCLUSION: Arsenic trioxide can effectively down-regulate the abnormal secretion of IL-10 and IFN-γ mRNA and influence the expression of Th factor cytokines, but not excessive immunosuppression and down-regulate the Th Abnormal proliferation of cells without affecting its survival rate, thereby regulating abnormal immune function in lupus mice may be one of the important mechanisms of arsenic in the treatment of SLE.