衰老标记蛋白30在糖尿病小鼠视网膜中的表达  

Expression of senescence marker protein 30 in retina of diabetic mice

作　　者：王洁[1] 李思雨 谢子康 左慧懿 唐承业[1] 唐东永[1] 忠昕 梁皓[1] WANG Jie;LI Siyu;XIE Zikang;ZUO Huiyi;TANG Chengye;TANG Dongyong;ZHONG Xin;LIANG Hao(Department of Ophthalmology,the First Affiliated Hospital of Guangxi Medical University,Nanning 530021,Guangxi Zhuang Autonomous Region,China)

机构地区：[1]广西医科大学第一附属医院眼科,广西壮族自治区南宁市530021

出　　处：《眼科新进展》2020年第3期216-220,共5页Recent Advances in Ophthalmology

基　　金：国家自然科学基金资助(编号81960174)。

摘　　要：目的从糖尿病与衰老之间生物学联系的角度探寻糖尿病视网膜病变(diabetic retinopathy,DR)早期干预的新靶点。方法从在线数据库Rat Genome Database(RGD)分别搜索与人类糖尿病和衰老有关的基因,通过STRING和GeneMANIA分别从蛋白和基因水平构建二者重叠基因的相互作用网络图,使用DAVID进行重叠基因的富集分析。用高脂饲料结合链-脲佐菌素(STZ)腹腔注射建立糖尿病小鼠模型,通过苏木精-伊红(HE)染色观察糖尿病小鼠视网膜结构变化,应用免疫组织化学法和RT-qPCR分别检测小鼠视网膜中上述筛选所得的新型衰老分子衰老标记蛋白30(senescence marker protein 30,SMP30)的蛋白及mRNA表达。结果通过RGD共筛选出与人类糖尿病和衰老共相关的包含新型衰老分子SMP30在内的基因11个,它们在蛋白和基因水平上都有着相互作用关系,富集分析显示其主要涉及神经元凋亡过程、凋亡过程负调控、NO生物合成过程的正调控、细胞衰老等;KEGG通路分析显示与线粒体生物发生和HIF-1信号通路相关。糖尿病小鼠视网膜各层排列不规则,其中内界膜不规则和神经节细胞排列错落紊乱尤为明显。对照组和糖尿病组SMP30蛋白分布于小鼠视网膜各层细胞胞浆且集中于神经纤维层、神经节细胞层、外丛状层,糖尿病组小鼠视网膜SMP30蛋白表达低于对照组(t=7.057,P<0.01)。RT-qPCR检测显示,糖尿病组小鼠视网膜中SMP30 mRNA的表达低于对照组(t=3.717,P=0.02)。结论糖尿病小鼠神经视网膜病理改变与SMP30表达降低有关,SMP30可作为DR早期的干预靶点,上调SMP30可能是保护视网膜神经组织免受糖尿病状态下胰岛素信号紊乱和氧化应激所致损伤的新途径。Objective To explore new targets for early intervention in diabetic retinopathy(DR)from the perspective of biological link between diabetes and aging.Methods Genes related to human diabetes and aging were searched from the online database Rat Genome Database(RGD).STRING and GeneMANIA were used to construct the interaction network diagrams of the overlapping genes from protein and gene levels respectively,and DAVID was used for enrichment analysis of the overlapping genes.Diabetic mouse model was established by intraperitoneal injection streptozotocin(STZ)as well as feeding with high fat diet.Changes in retinal structure of diabetic mice were observed by hematoxylin-eosin(HE)staining,expression of SMP30 protein and mRNA in mice retina were respectively detected by immunohistochemistry and RT-qPCR.Immunohistochemistry and RT-qPCR were used to detect the protein and mRNA expression of senescence marker protein 30(SMP30),a novel senescence molecule obtained from the retina in the mouse.Results RGD was used to screen totally 11 genes co-related to human diabetes and aging,which included new aging molecule SMP30,and they had interactions at the protein and gene levels.Enrichment analysis showed that they mainly involved neuronal apoptosis,negative regulation of apoptosis,positive regulation of nitric oxide biosynthesis and cellular senescence;KEGG pathway analysis had been shown to be related to mitochondrial biogenesis and HIF-1 signaling pathway.Retinal layers of diabetic mice were irregularly arranged,and irregularity of inner limiting membrane and disorder of the ganglion cells were particularly obvious.SMP30 proteins in control group and diabetic group were distributed in the cytoplasm of retinal layers and concentrated in nerve fiber layer,ganglion cell layer and outer plexiform layer in each group.But expression of SMP30 protein in retina of diabetic group was lower than that in control group(t=7.057,P<0.01).RT-qPCR showed that the expression of SMP30 mRNA in the retina of diabetic mice was lower than that in th

关 键 词：衰老标记蛋白30 糖尿病 衰老 神经视网膜 

分 类 号：R774[医药卫生—眼科]