利拉鲁肽改善胰岛素抵抗大鼠的肾脏氧化应激作用  

Liraglutide improves renal oxidative stress in rats with insulin resistance

作　　者：吴明昊[1] 刘剑[1] 高宇[3] 胡桂才[2] 张宝红[2] 周瑾[2] 赵亚娟[2] 

机构地区：[1]承德医学院,067000 [2]承德医学院附属医院肾脏内科 [3]承德医学院附属医院内分泌科

出　　处：《中华糖尿病杂志》2016年第9期559-563,共5页CHINESE JOURNAL OF DIABETES MELLITUS

摘　　要：目的：观察利拉鲁肽对高脂诱导的胰岛素抵抗（IR）大鼠肾功能、电镜下肾小球形态及肾脏还原型烟酰胺腺嘌呤二核苷酸磷酸氧化酶P22phox、P47phox表达的影响。方法将54只雄性Wistar大鼠分为普通饲料组（16只）和高脂饲料组（38只），喂养8周末各组取5只行高胰岛素-正葡萄糖钳夹试验，测定葡萄糖输注率（GIR），判定IR大鼠造模成功。将高脂饲料组大鼠按随机数字表法分为3组：高脂组（B组）、利拉鲁肽100μg/（kg·d）组（C组）、利拉鲁肽200μg/（kg·d）组（D组），各组11只，普通饲料组为对照组（A组，n=11）。A、B两组每天生理盐水皮下注射，C、D组药物干预2周。各组取5只大鼠再行钳夹试验计算GIR，并分别测定各组大鼠生化指标及肾组织P22phox、P47phox mRNA及蛋白表达水平，透射电镜观察大鼠肾脏超微结构。两组间比较采用LSD t检验，多组间比较采用单因素方差分析。结果（1）与A组比较，B组P22phox、P47phox的mRNA及蛋白表达水平显著上升（P22phox mRNA分别为4.9±0.4比1.0±0.0，P22phox蛋白2.72±0.20比1.27±0.24；P47phox mRNA 4.8±0.8比1.0±0.0，P47phox蛋白2.17±0.39比1.13±0.15，t=16.041、8.040、7.914、4.225；均P〈0.05）。（2）与B组比较，C、D组P22phox、P47phox mRNA及蛋白表达水平下降（P22phox mRNA在D、C、B组分别为2.3±0.4、4.5±0.4、4.9±0.4，P22phox蛋白1.75±0.13、2.32±0.21、2.72±0.20；P47phox mRNA在D、C、B组分别为2.2±0.3、3.6±0.4、4.8±0.8，P47phox蛋白为1.30±0.18、1.66±0.15、2.17±0.39），且D组比C组下降更为显著（t=6.482、3.401、4.875、2.067，均P〈0.05）。（3）B组肾小球基底膜厚薄不均，部分足突肿胀、融合甚至消失。经利拉鲁肽干预后有所减轻，且D组改善效果更为明显。结论通过抑制肾脏P22phox、P47phox的表达，减轻肾脏氧化应激，这可能是利拉鲁肽保护IR大鼠肾脏的作用机制之一。Objective To investigate the effects of liraglutide on renal function, the glomerular morphology in electron microscope and oxidase subunit of dihydronicotinamide adenine dinuclectide phosphate（NADPH） —P22phox and P47phox in kidney in rats with insulin resistance（IR）. Methods Fifty-four male Wistar rats were divided into normal diet group（group A, n=16） and high-fat diet group（n=38）. Hyperinsulinemic-euglycemic clamp test was performed at the waking state after 8 weeks feeding. Glucose infusion rate（GIR） was measured to determine if IR rats model was successful. The rats with high-fat diet were randomly divided into three groups：high-fat diet group（group B, n=11）, liraglutide 100μg/（kg · d） group （group C, n=11）, liraglutide 200μg/（kg·d） group（group D, n=11）. The rats with normal diet was group A（n=11）. Daily subcutaneous injections of saline were given group A, B,while liraglutide intervention was given in group C,D. After 2 weeks, biochemical markers were measured in each group. Realtime Quantitative PCR and Western blotting were used for detecting the expression of P22phox and P47phox in kidney. Renal ultrastructure was observed by transmission electron microscope. LSD t test was performed in comparison between the two groups, while one way variance analysis for multiple groups comparison. Results （1） Compared with group A, the expression of P22phox and P47phoxin were significantly increased in group B （P22phox mRNA：4.9±0.4 vs 1.0±0,P22phox protein：2.72±0.20 vs 1.27±0.24;P47phox mRNA：4.8±0.8 vs 1.0 ± 0,P47phox protein： 2.17 ± 0.39 vs 1.13 ± 0.15,t=4.225-16.041, all P〈0.05）. （2） Compared with the group B, the expression level of P22phox and P47phox were decreased in liraglutide groups （P22phox mRNA： 2.3 ± 0.4, 4.5 ± 0.4, 4.9 ± 0.4 in group D,C,B, respectively; P22phox protein 1.75 ± 0.13, 2.32 ± 0.21, 2.72 ± 0.20;P47phox mRNA： 2.2 ± 0.3, 3.6 ± 0.4, 4.8 ± 0.8,in group D, C, B, respectively; P47phox protein：1.30±0.

关 键 词：胰岛素抵抗 氧化应激 烟酰胺腺嘌呤二核苷酸氧化酶 肾脏 利拉鲁肽 

分 类 号：R587.1[医药卫生—内分泌]