α-胞衬蛋白siRNA对干燥综合征模型NOD小鼠IFN-γ和IL-4表达水平及基因沉默效果的影响  

Effects of α-Fodrin siRNA on expressions of IFN-γ and IL-4 and gene silencing of Sjogren's syndrome model NOD mice

作　　者：庞春艳[1] 吕凤凤[1] 杨麟[1] 王永福[1] 

机构地区：[1]内蒙古科技大学包头医学院风湿免疫研究所,内蒙古包头014010

出　　处：《吉林大学学报（医学版）》2013年第3期548-553,F0003,共7页Journal of Jilin University：Medicine Edition

基　　金：内蒙古自治区教育厅高等学校科学技术研究项目资助课题(NJ09165)

摘　　要：目的:观察α-胞衬蛋白(α-Fodrin)特异性小干扰RNA(siRNA)对干燥综合征(SS)动物模型非肥胖糖尿病(NOD)小鼠细胞因子IFN-γ和IL-4表达水平的影响,探讨α-Fodrin siRNA治疗NOD的可能性。方法:12只NOD雌性小鼠随机分为对照组、空载体组、α-Fodrin siRNA1组和α-Fodrin siRNA2组,每组3只。将成功构建的α-Fodrin siRNA1和α-Fodrin siRNA2表达载体分别经尾静脉注射α-Fodrin siRNA1组和α-Fodrin siRNA2组NOD小鼠,对照组和空载体组小鼠分别注射同等剂量的PBS和pGFP-V-RS空载体,测定每周每只小鼠的饮水量。注射后1、3、5和7d,各组小鼠尾部采血,ELISA法检测4组小鼠血清中细胞因子IFN-γ、IL-4的表达水平,RT-PCR法检测NOD小鼠肺脏组织中α-Fodrin mRNA的表达水平,免疫组织化学法检测NOD小鼠肺脏组织中α-Fodrin的表达水平。结果:4组小鼠饮水量比较差异无统计学意义(P>0.05)。注射后1和3d,siRNA1组和siRNA2组NOD小鼠血清中IL-4水平明显升高、IFN-γ/IL-4比值减低,与对照组和空载体组比较差异有统计学意义(P<0.05),对照组与空载体组比较差异无统计学意义,siRNA1组与siRNA2组比较差异亦无统计学意义;注射后5和7d,4组小鼠血清中IL-4水平和IFN-γ/IL-4比值比较差异无统计学意义。注射后7d,siRNA1组和siRNA2组NOD小鼠肺脏组织中α-Fodrin及其mRNA的表达水平明显低于对照组和空载体组(P<0.05),对照组和空载体组比较差异无统计学意义,siRNA1组和siRNA2组比较差异无统计学意义。结论:NOD小鼠注射α-Fodrin siRNA载体后,血清中IL-4的水平升高、IFN-γ/IL-4的比值降低,同时α-FodrinmRNA表达水平和α-Fodrin表达水平下降,提示α-Fodrin siRNA可以通过减轻NOD小鼠炎症反应延缓SS病情进程。Objective To observe the effects of α-Fodrin siRNA on the expressions of IFN-γ and IL-4 of Sjogren＇s syndrome（SS） model non-obese diabetic（NOD） mice and to discuss its possibility on treatment of NOD.Methods 12 female NOD mice were randomly divided into control group（n=3）,empty vector group（n=3）,α-Fodrin siRNA1 group（n=3）,and α-Fodrin siRNA2 group（n=3）.The vectors expressing α-Fodrin and siRNA 1 and α-Fodrin siRNA2 were injected by tail vein into NOD mice in α-Fodrin siRNA1 group and α-Fodrin siRNA2 group,while the NOD mice in control group and empty vector group were injected with the same dose of PBS and pGFP-V-RS empty vector,respectively.The water drinking amount of each mouse was detected in each week.The expression levels of IFN-γ and IL-4 in serum of the mice in various groups were detected by ELISA method on the 1st,3rd,5th,and 7th day after injection,and the expression levels of α-Fodrin mRNA in lung tissue of NOD mice were detected by RT-PCR method and the expression level of α-Fodrin protein was detected by immunohistochemistry method.Results There were no significant differences of water drinking amounts of the mice between four groups（P0.05）.The levels of IL-4 were higher and IFN-γ/IL-4 ratios were lower in siRNA1 group and siRNA2 group than control group and empty vector group on the 1st and 3rd day after injection（P0.05）;the levels of IL-4 and IFN-γ/IL-4 in control group and empty vector group had no significant differences on the 1st and 3rd day after injection;so did α-Fodrin siRNA1 group and α-Fodrin siRNA2 group.There were no significant differences of the levels of IL-4 and the ratios of IFN-γ/IL-4 between various groups on the 5th and the 7th day.The expression levels of α-Fodrin mRNA and protein were lower in siRNA1 group and siRNA2 group than control group and empty vector group on the 7th day after injection（P0.05）;the expression levels of α-Fodrin protein were lower in siRNA1 group and siRNA2 than control group and empty vector group

关 键 词：Α-胞衬蛋白 小干扰RNA 非肥胖糖尿病 白细胞介素4 干扰素γ 干燥综合征 

分 类 号：R587[医药卫生—内分泌]