TLR4抑制剂TAK-242对高脂饮食诱导的小鼠胰岛素抵抗的干预作用  

Intervention role of TLR4 inhibitor TAK-242 on insulin resistance states in mice with a high fat diet-induced insulin resistance

作　　者：王振辉[1,2] 马科[2] 朱晓波[2] 郝敏[2] 常晓彤[2] 

机构地区：[1]解放军第251医院核医学科,张家口075000 [2]河北北方学院临床检验诊断学重点实验室,张家口075000

出　　处：《免疫学杂志》2016年第11期928-934,共7页Immunological Journal

基　　金：河北省自然科学基金(C2011405015);河北北方学院创新人才培育项目(CXRC1316)

摘　　要：目的建立高脂饮食诱导的胰岛素抵抗小鼠模型,探讨干预Toll样受体4(Toll-like receptor 4,TLR4)对高脂饮食诱导的小鼠胰岛素抵抗状态的影响。方法首先建立高脂饮食诱导的胰岛素抵抗小鼠模型。选取出生21 d的雄性C57BL/6小鼠36只,随机分为2组,正常对照组12只,以普通基础饲料喂养(low fat diet,LFD),高脂饮食实验组24只,给予高脂饲料喂养(high fat diet,HFD)。待2组小鼠体质量出现显著差异时,进行葡萄糖耐量实验(glucose tolerance test,GTT)和胰岛素耐量实验(insulin tolerance test,ITT)观察小鼠胰岛素抵抗的发生。小鼠胰岛素抵抗模型建立后,观察TLR4抑制剂TAK-242对小鼠胰岛素抵抗状态的作用。LFD组小鼠继续以基础饲料喂养(即作LFD对照组);HFD组小鼠随机分为2组,每组12只小鼠,均继续以高脂饲料喂养,其中一组HFD小鼠腹腔注射TLR4抑制剂TAK-242,以0.5 mg TAK-242/kg体质量的计量给予,每周2次(即给予TAK-242的高脂饮食实验组,HFD-T组);另一组HFD小鼠作为单纯高脂饮食的胰岛素抵抗空白对照组(HFD-C组),只给予等体积的溶媒-二甲基亚砜(DMSO)。给予小鼠TLR4抑制剂5个月,进行GTT和ITT后,处死小鼠。采用EDTA-K2抗凝管收集血液标本,立即分离外周血单核细胞(peripheral blood mononuclear cell,PBMC)和血浆。应用Western blot技术检测PBMC TLR4蛋白表达水平。采用全自动生化分析仪测定血浆葡萄糖(GLU)、甘油三酯(TG)、总胆固醇(TC)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、丙氨酸氨基转移酶(ALT)的水平。结果在小鼠胰岛素抵抗模型建立期间,与对照组相比,高脂饮食组小鼠体质量增长较快,喂养至第20周体质量出现显著差异(P<0.05);持续高脂饮食7个月,GTT和ITT结果显示,高脂饮食组小鼠对糖的调节能力受损、对胰岛素的降糖作用减低,说明高脂饮食诱导小鼠胰岛素抵抗成功。出现胰岛素抵抗的小鼠,在给予TLR4抑制剂The role of Toll-like receptor 4（TLR4） in obesity-related insulin resistance is not well known.This study aimed to investigate the intervention role of TLR4 inhibitor TAK-242 on insulin resistance states in mice with a high fat diet-induced insulin resistance.In order to achieve this aim,a high fat diet-induced insulin resistance model was established in mice.A total of 36 male C57BL/6 mice（born 21 days） were selected and randomly divided into two groups：the control group（12 mice） was fed with general basic diet（low fat diet,LFD）,while the experimental group（24 mice） was fed with high fat diet（high fat diet,HFD）.When the two groups demonstrated significant differences in body weight,the glucose tolerance test（GTT） and insulin tolerance test（ITT） were performed for observing the occurrence of insulin resistance in mice.After the mouse model of insulin resistance was established,the role of TLR4 inhibitor TAK-242 in insulin resistance was studied.LFD mice were still fed with general basic diet（LFD control group）;HFD mice were randomly divided into 2 groups,with 12 mice in each group,and fed with high fat diet.One group of HFD mice was injected intraperitoneally with TLR4 inhibitor TAK-242（0.5 mg TAK-242/kg body weigh）,2 times a week（HFD-T group）;another group of HFD mice was designed as control group（HFD-C group）.LFD control group and HFD-C group mice were only injected with an equal volume Dimethyl sulfoxide（DMSO）.After been giving mouse TLR4 inhibitors for GTT and ITT for 5months,the mice were sacrificed.The blood samples were collected using EDTA-K_2 anticoagulant tubes,and then peripheral blood mononuclear cells（PBMC） and plasma were isolated immediately.Western blot was applied to detect PBMC TLR4 protein levels.Automatic biochemical analyzer was used to detect plasma glucose（GLU）,triglyceride（TG）,total cholesterol（TC）,high-density lipoprotein cholesterol（HDL-C）,low-density lipoprotein cholesterol（LDL-C）,alanine aminotransferase（AL

关 键 词：高脂饮食 胰岛素抵抗 肥胖 TOLL样受体4 抑制剂TAK-242 

分 类 号：R589.2[医药卫生—内分泌]