三氧化二砷对MRL/lpr狼疮鼠T-bet/GATA3信号通路的影响  

Effects of arsenic trioxide on T-bet/GATA3 signal pathway in MRL/lpr mice

作　　者：朱晓芳[1] 胡宏叶 李永吉[1] 朱小春[1] 张章 陈丹[1] 

机构地区：[1]温州医科大学附属第一医院风湿免疫科,浙江温州325000 [2]温州医科大学附属第一医院肿瘤外科,浙江温州325000

出　　处：《中国病理生理杂志》2017年第9期1708-1712,共5页Chinese Journal of Pathophysiology

基　　金：温州科技局重大项目(No.Y20090240)

摘　　要：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、Tbet/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Tbet/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。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 in2 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 from 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-q PCR. 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-γ and 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.Table 1. The sequences of the primers for RT-q PCR Name The sequence of the primer（5＇-3＇） Product（ bp）T-bet Forward： gtttgtgcatctccttccatct 148Reverse： ggaggtggtagcctttaatcct GATA3 Forward： ccaggcaagatgagaaagagtg 149Reverse： atagggcggataggtggtaatg IFN-γ Forward： ggaaccctctcccttcaatgt 136Reverse： ctccacaatagccttcagtgc IL-4 Forward： gaactcagacccacccaaaag 141Reverse： tacacaataccaaagcccaggtβ-actin Forward： gagaccttcaacaccc

关 键 词：三氧化二砷 MRL/lpr狼疮鼠 T-bet/GATA3信号通路 IFN-γ IL-4 

分 类 号：R593.2[医药卫生—内科学]