三氧化二砷对MRL/lpr狼疮鼠T-bet/GATA3信号通路的影响

摘要

AIM:To investigate the effect of arsenic trioxide (ATO) on T-bet/GATA3 signal pathway in MRL/lpr mice.METHODS:MRL/lpr mice and C57BL/6J mice at the age of 20 weeks were chosen and then divided in 2 different sub-groups,respectively.The mice in 2 sub-groups received ATO (0.4 mg · kg-1.d-1) and sodium chloride (NS,volume weight-determined) by intraperitoneal injection respectively for 2 months.Afterward,the spleens were isolated fron the MRL/lpr and C57BL/6J mice under pathogen-free condition and the suspensions were prepared.The mRNA level of T-bet,GATA3,IFN-γ,IL-4 and the mRNA ratio of T-bet/GATA3 were detected by RT-qPCR.The protein expression of T-bet and GATA3 was determined by Western blot.The serum levels of IFN-γ and IL-4 were measured by ELISA.RESULTS:The mRNA and protein levels of T-bet,IFN-γand the mRNA ratio of T-bet/GATA3 in NS group of MRL/lpr mice were higher than those in NS group of C57BL/6J mice (P ＜0.05).However,the GATA3 and IL-4 were lower in NS group of MRL/lpr mice in both mRNA and protein level (P ＜ 0.05).In MRL/lpr mice,the mRNA and protein levels of T-bet,IFN-γ and the mRNA ratio of T-bet/GATA3 were lower in ATO group compared with NS group (P ＜ 0.05),no difference was found in GATA3 and IL-4.No difference of the indexes mentioned above between ATO group and NS group in C57BL/6J mice was observed.CONCLUSION:ATO may affect the signaling pathway of T-bet/GATA3 to down-regulate the mRNA expression and the protein secretion of IFN-γ by decreasing the expression of T-bet in MRL/lpr mice.%目的:本文旨在探讨三氧化二砷(arsenic trioxide,ATO)对MRL/lpr狼疮鼠T-bet/GATA3信号通路的影响及其作用机制.方法:将20周龄MRL/lpr狼疮鼠和正常C57BL/6J小鼠随机分组,分别接受ATO(0.4mg·kg-1.d-1)和同体积生理盐水(NS)治疗2个月,分离脾脏淋巴细胞,然后用实时荧光定量PCR法检测T-bet、GATA3、IFN-γ、IL-4的mRNA水平及T-bet/GATA3 mRNA的比值,Western blot法检测T-bet和GATA3的蛋白表达,ELISA法检测血清中IFN-γ和IL-4的浓度.结果:(1)MRL/lpr狼疮鼠NS组T-bet、IFN-γ的mRNA及蛋白表达、T-bet/GATA3 mRNA比值均高于C57BL/6J小鼠NS组(P＜0.05),而GATA3、IL-4的mRNA及其蛋白表达均低于C57BL/6J小鼠NS组(P＜0.05);(2)在MRL/lpr狼疮鼠中,与NS组相比,ATO组T-bet、IFN-γ的mRNA及蛋白表达、T-bet/GATA3 mRNA比值均下降(P＜0.05),而2组中GATA3、IL-4的mRNA及蛋白表达无明显差异;(3)在C57BL/6J小鼠中,NS组和ATO组之间以上指标均无显著性差异.结论:ATO可能通过影响MRL/lpr狼疮鼠T-bet/GATA3信号通路,降低T-bet表达,从而减少Th1型细胞因子IFN-γ的表达和分泌.