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

发布时间：2018-01-09 22:26

本文关键词：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　出处：《吉林大学学报(医学版)》2013年03期 　论文类型：期刊论文

【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(P0.05)。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(P0.05),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(P0.05),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。
[Abstract]:Objective: To observe the effect of alpha fodrin (alpha -Fodrin) specific small interfering RNA (siRNA) of Sjogren syndrome (SS) animal model of non obese diabetic (NOD) mice expression of cytokines IFN- and IL-4, to explore the possibility of a -Fodrin siRNA for the treatment of NOD. Methods: 12 NOD female mice were randomly divided into control group, empty vector group, siRNA1 group and alpha -Fodrin alpha -Fodrin siRNA2 group, 3 rats in each group. The successful construction of the -Fodrin siRNA1 alpha and alpha -Fodrin expression vector siRNA2 respectively by intravenous injection of alpha -Fodrin alpha -Fodrin siRNA2 siRNA1 group and NOD group, PBS and pGFP-V-RS were injected with the same dose of vector light group and vector group mice, each mouse was measured weekly. The amount of water after injection of 1,3,5 and 7d, the mice tail blood ELISA cytokines were measured by 4 groups of mice serum levels of IFN-, the expression level of IL-4 and RT-PCR were detected in lung tissue of NOD mice The expression level of alpha -Fodrin mRNA, detection of NOD expression in mouse lung tissue by immunohistochemistry in alpha -Fodrin. Results: there was no significant difference between the 4 groups of mice drinking water (P0.05). After the injection of 1 and 3D, IL-4 level of siRNA1 group and siRNA2 group NOD in the serum of mice was significantly increased, the ratio of /IL-4 IFN- gamma reduced, there was statistically significant difference compared with the control group and empty vector group (P0.05), the control group and empty vector group had no significant difference, the difference between the siRNA1 group and siRNA2 group have no statistical significance; after the injection of 5 and 7d, no significant difference was found in the level of IFN- and the ratio of /IL-4 gamma IL-4 4 groups in the serum of mice difference. After injection of 7D, the expression level of -Fodrin and mRNA alpha siRNA1 group and siRNA2 group of NOD mice lung tissues was significantly lower than the control group and empty vector group (P0.05), the control group and empty vector group but no significant difference was found between group siRNA1 and S The difference between the iRNA2 group had no statistical significance. Conclusion: NOD mice were injected with alpha -Fodrin siRNA vector, elevated serum IL-4 level, IFN- ratio of gamma /IL-4 decreased, while the expression of -FodrinmRNA alpha and alpha -Fodrin expression level decreased, suggesting that siRNA alpha -Fodrin can reduce NOD mice inflammatory response delay SS disease process.

【作者单位】：内蒙古科技大学包头医学院风湿免疫研究所;
【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)
【分类号】：R589;R-332
【正文快照】： 干燥综合征(Sjogren’s syndrome,SS)是一种以泪腺、涎腺等外分泌腺受累为主要特征的系统性自身免疫性疾病。多种细胞因子和自身抗体参与了SS的致病过程[1-3]。α-胞衬蛋白(α-Fodrin)是从干燥小鼠唇腺中提取的一种相对分子质量为120 000的涎腺特异性自身抗原。研究[4]表明:α

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