Toll样受体4对2型糖尿病小鼠视网膜病变的影响  

Effects of TLR4 on retinopathy of typeⅡdiabetes mice

作　　者：张经[1] 肖青[2] 姜发纲[2] 刘栋 赵曼[1] 

机构地区：[1]西安市第一医院眼科,陕西省眼科研究所,陕西省西安市710002 [2]武汉协和医院眼科,湖北省武汉市453000 [3]同济医院放射科,湖北省武汉市430022

出　　处：《眼科新进展》2016年第9期805-808,共4页Recent Advances in Ophthalmology

基　　金：国家自然科学基金资助(编号:30801266)~~

摘　　要：目的探讨Toll样受体4(toll-like receptor 4,TLR4)对2型糖尿病小鼠模型糖尿病视网膜病变(diabetic retinopathy,DR)的影响。方法取16周龄的健康雄性TLR4基因缺陷小鼠C3H/He J(mutation,Mut)及相应野生型小鼠C3H/He N(wild type,WT)各12只,用链脲佐菌素(STZ)腹腔注射建立2型糖尿病动物模型,另取4只未处理小鼠做对照,分为Mut组、WT组与对照组。于模型建成后1个月、2个月、4个月分别取材,每个时间点模型组分别取材4只,对照组分别取16周龄的两种小鼠各2只眼球。通过电镜观察小鼠视网膜组织超微结构的变化,并应用RTq PCR检测视网膜TLR4及巨噬细胞炎性蛋白2(macrophage inflammatory protein-2,MIP-2)的mRNA表达。结果糖尿病模型造模后1个月的超微电镜显示,小鼠视网膜组织即有神经节细胞及神经纤维水肿,核周间隙增宽,核固缩,胞浆颜色变淡;模型造模后2个月时毛细血管内皮细胞胞浆线粒体水肿,嵴变短,周细胞水肿;模型造模后4个月时基底膜节段性增厚,微绒毛突起,管腔狭窄,各时间点的病变WT组均重于Mut组。视网膜组织RT-q PCR结果显示,WT组和Mut组TLR4及MIP-2的mRNA的表达在糖尿病模型造模后1个月、2个月、4个月时依次上调(均为P<0.001),各时间点TLR4及MIP-2的mRNA的表达以WT组强于Mut组(均为P<0.05)。结论 2型糖尿病小鼠模型中,随病程发展出现视网膜病变并不断恶化,此过程中视网膜中TLR4与MIP-2的mRNA表达呈显著性上调趋势;TLR4基因突变小鼠的视网膜TLR4及MIP-2的表达比野生型小鼠减弱,同时视网膜病变减轻。Objective To investigate the effects of Toll-like receptor 4 （TLR4） on the diabetic retinopathy （DR） in type ]1 diabetic mice. Methods Twenty healthy male TLR4 mutant （C3H/HeJ;Mut） mice and 12 healthy male wild-type mice （C3H/ HeN;WT） withl5 weeks old were established the diabetic model with STZ injection, and other 4 normal mice were used as control group. At 1 month,2 months and 4 months after the models establishment, the mice were sacrificed, the eyeballs were enucleated, and 4 cases at each time point. The retinal microvascular ultrastructure was observed under transmission electron microscope, the expressions of TLR4 and macrophage inflammatory protein 2 （ MIP-2 ） mRNA in the retina were detected by RT-qPCR. Results At 1 month after the models establishment, the transmission electron microscope showed that the ganglion cells and nerve fiber were edema, the nuclear space was increased with nuclear condensation;At 2 month after the models establishment, the mitochondria of capillary endothelial cell were swollen, the crista was shorten and disappeared, and the peripheral cells were edema; At 4 month after the models establishment,the basement membrane of capillaries became thicker, microvilli became tuber, and the capillary lumen became narrowing;At each time point,the lesions of WT group was more serious than Mut group. RT-qPCR showed that the expression of MIP-2 and TLR4 mRNA in the retina was up-regulated in the model groups with the course of DR prolonged （P 〈 0. 000 1 ）, at each time point the expression of WT group was stronger than Mut group （ all P 〈 0.05 ）. Conclusion The retinopathy is more and more serious with the course prolong in type Ⅱ diabetic mice model, and at the same time the expression of MIP-2 and TLR4 mRNA in the retina are up-regulated; The expressions of MIP-2 and TLR4 mRNA in the TLR4 mutant mice are weaker than those in the wild-type mice, and the lesion is weaker meanwhile.

关 键 词：TOLL样受体4 2型糖尿病视网膜病变 巨噬细胞炎性蛋白2 

分 类 号：R774[医药卫生—眼科]