转基因过表达IL-10对NOD小鼠胰岛形态和功能的保护及其机能的观察  

Protective effect of gene transfer-induced IL-10 over-expression on morphology and function of pancreatic beta cells

作　　者：徐爱晶 陈志红[2] 阎丽华[3] 田飞[2] 李堂[2] 

机构地区：[1]广州市妇女儿童医疗中心内分泌代谢科,510520 [2]青岛大学医学院附属医院儿科 [3]青岛市妇女儿童医疗保健中心

出　　处：《中国糖尿病杂志》2013年第7期625-629,共5页Chinese Journal of Diabetes

基　　金：国家自然科学基金(81170762);广东省自然科学基金(S2012040006330)

摘　　要：目的观察转基因方法诱导IL-10过表达对NOD小鼠胰岛形态和功能的保护及其机能。方法 8周龄NOD小鼠随机分为腺病毒介导IL-10(Ad-IL-10)组、腺病毒空载体Ad-GFP(Ad-GFP)组及NC组。检测各组胰腺IL-10的表达,外周血胰岛素水平;观察胰腺组织形态变化;检测NOD小鼠血清IL-10、TNF-α、IFN-γ水平;测定胰岛细胞凋亡及Fas、Caspase-3的表达。结果腹腔注射Ad-IL-10可成功转染NOD小鼠胰腺细胞,减少糖尿病累积发病率,增加胰岛素水平、减轻胰岛炎。TNF-α、IFN-γ水平在Ad-IL-10组低于NC组及Ad-GFP组(P<0.01);Ad-IL-10组胰岛β细胞凋亡率低于Ad-GFP组及NC组(14.1%vs 32.8%vs 35.4%,P<0.01);Fas、Caspase-3蛋白在Ad-IL-10组表达较其他两组降低(P<0.01)。结论 IL-10在NOD小鼠胰腺组织过表达可以减轻胰岛炎、减少糖尿病的发生,通过Fas途径减少细胞凋亡。Objective To explore the effect and mechanism of IL-10 gene transfer on morphology and function of pancreatic beta cells in NOD mice. Method Female NOD mice, aged 8 weeks, were randomized into 3 groups： normal control group, Ad-GFP group and Ad-IL-10 group. Before and during the experiment, the blood glucose of all mice was assessed. Panereata were removed from NOD mice at the 16th week of age in each group and their insulitis severity was scored by routine FIE staining. The levels of IL-10, TNF-α, IFN-7 and insulin in sera were measured by the ELISA method. The apoptosis of pancreatic β-cells was determined by TUNEL assay and the expressions of Fas and easpase-3 were estimated by immunohistochemistry analysis. Results There was a significant reduction of diabetes incidence in NOD mice treated with IL-10 gene. The instis score of the Ad-IL-10 group was lower than that of the normal control group and Ad-GFP group （P〈0. 01）. The sera level of IL-10 was higher and TNF-α and IFN-γ were lower in the Ad-IL-10 group than those in the normal control group （P〈0. 01）. The apoptosis of fl cells was lower in the Ad-IL-10 group than in the Ad-GFP group and the normal control group （14. 1% vs 32. 8 % vs 35. 4 0%, P〈0. 01） by TUNEL assay. The expressions of Fas and easpase- 3 were decreased in the Ad-IL-10 group than in the other two groups （P〈0. 01）. Conclusion The gene transfer-induced IL-10 over-expression alleviates the insulitis of NOD mice, reduces the incidence of diabetes, and reduces the apoptosis via decreasing the expression of Fas.

关 键 词：IL-10 基因 凋亡 NOD小鼠 

分 类 号：R587.1[医药卫生—内分泌]