前列腺素E2及其受体在链脲佐菌素诱导的糖尿病大鼠视网膜病变中的作用及机制研究  

Role and Mechanism of Prostaglandin E2 and Its Receptor in the Retinopathy in Diabetic Rats Induced by Streptozotocin

作　　者：车选义[1] 杨燕[2] 石蕊[1] 李春花[3] 杨文[3] 张倩[3] CHE Xuan-yi;YANG Yan;SHI Rui;LI Chun-hua;YANG Wen;ZHANG Qian(Department of Ophthalmology,Shaanxi Provincial People's Hospital,Affiliated Hospital of Xi'an Medical University,Xi'an,Shaanxi,710068,China;Operating Room,Shaanxi Provincial People's Hospital,Xi'an,Shaanxi,710068,China;Department of Ophthalmology,Xi'an Fourth Hospital,Xi'an,Shaanxi,710004,China)

机构地区：[1]陕西省人民医院西安医学院附属医院眼科,陕西西安710068 [2]陕西省人民医院手术室,陕西西安710068 [3]西安市第四医院眼科,陕西西安710004

出　　处：《现代生物医学进展》2020年第22期4218-4224,共7页Progress in Modern Biomedicine

基　　金：陕西省重点研发计划项目(2017SF-249)。

摘　　要：目的:探究前列腺素E2(prostaglandin E2,PGE2)及其受体前列腺素E2受体(E-prostanoid2 receptor,EP2R)在链脲佐菌素诱导的糖尿病大鼠视网膜病变中的作用及机制。方法:将SD大鼠随机分为6组:对照组使用标准饲料喂养;其他组大鼠使用高脂高糖饲料喂养+腹膜内注射链脲佐菌素(30 mg/kg)建立糖尿病大鼠模型;PGE2组、Butaprost组、AH6809组大鼠分别给予玻璃体腔内注射5 mmol/L的PGE2、EP2R激动剂Butaprost或EP2R抑制剂AH6809,注射剂量为6μL。DMSO组注射等剂量DMSO盐溶液。每周注射1次,共注射4周。通过苏木精伊红(HE)染色评价视网膜病变;免疫组化或蛋白质印迹分析视网膜组织中EP2R、胰岛素受体底物1(IRS-1)、磷酸肌醇3激酶(PI3K)、p-PI3K、蛋白激酶B(Akt)、p-Akt、细胞间粘附分子-1(ICAM-1)、内皮一氧化氮合酶(eNOS)、核因子κB p65(NF-κB p65)和血管内皮生长因子(VEGF)的表达。此外,分别应用PGE2、Butaprost或AH6809处理高糖培养基(4.5 g/L葡萄糖)培养的视网膜微血管内皮细胞系(HRMEC),并检测各组细胞活力、细胞凋亡率和血管生成情况。结果:与正常视网膜组织相比,糖尿病大鼠视网膜组织中EP2R呈显著高表达(P<0.05)。与对照组和模型组相比,PGE2和Butaprost组的EP2R、IRS-1、p-PI3K、p-Akt、ICAM-1、eNOS、NF-κBp65和VEGF的表达水平显著升高,而AH6809组的上述蛋白的表达水平显著降低(P<0.05)。体外研究中,与对照组和模型组HRMEC相比,PGE2和Butaprost处理的HRMEC的活力和血管生成数量显著升高,而细胞凋亡率显著降低,AH6809处理则抑制了上述细胞改变(P<0.05)。结论:PGE2/EP2R可能通过促进IRS-1/PI3K/Akt信号通路介导的炎症反应、细胞凋亡和血管生成促进糖尿病视网膜病变的发生和发展。Objective:To investigate the role and mechanism of prostaglandin E2(PGE2)and its receptor E-prostanoid2 receptor(EP2 R)in retinopathy induced by streptozotocin in diabetic rats.Methods:SD rats were randomly divided into 6 groups:the control group was fed with standard feed;the other groups were fed with high-fat and high-sugar feed+intraperitoneal injection of streptozotocin(30 mg/kg)to establish a diabetic rat model.The rats in PGE2 group,Butaprost group,and AH6809 group were injected intravitreally with 5 mmol/L of PGE2,EP2 R agonist Butaprost or EP2 R inhibitor AH6809,with a volume of 6μL.The DMSO group was injected with an equal volume of DMSO salt solution.Inject once a week for 4 weeks.Retinopathy was evaluated by hematoxylin and eosin(HE)staining.The expression of EP2 R,Insulin receptor substrate-1(IRS-1),Phosphoinositide 3-kinase(PI3 K),p-PI3 K,Protein kinase B(Akt),p-Akt,Intercellular adhesion molecule-1(ICAM-1),Endothelial nitric oxide synthase(eNOS),Nuclear factor kappa-B p65(NF-κB p65),Vascular endothelial growth factor(VEGF)in retinal tissues was assessed by immunohistochemical or western blot analysis.In addition,PGE2,Butaprost or AH6809 were used to treat the retinal microvascular endothelial cell line(HRMEC)cultured in high glucose medium(4.5 g/L glucose),and the cell viability,apoptosis rate and angiogenesis were detected.Results:Compared with normal retinal tissue,EP2 R was abnormally highly expressed in retinal tissue of diabetic rats(P<0.05).Compared with the control group and the model group,the expression levels of EP2 R,IRS-1,p-PI3 K,p-Akt,ICAM-1,eNOS and NF-κBp65 in PGE2 and butaprost groups were significantly higher,while those in ah6809 group were significantly lower(P<0.05).In vitro studies,compared with the control group and model group HRMEC,the activity and number of angiogenesis of PGE2 and Butaprost-treated HRMEC were significantly increased,while the apoptosis rate was significantly reduced.AH6809 treatment inhibited the above cell behavior(P<0.05).Conclusions:Inhibition of PGE

关 键 词：糖尿病性视网膜病变 前列腺素E2 前列腺素E2受体 视网膜微血管内皮细胞 IRS-1/PI3K/Akt信号通路 

分 类 号：R-33[医药卫生] R774.1