摘要
目的探讨甲酰肽受体Fpr2在氧诱导视网膜病变(OIR)小鼠中对胶质细胞改变的影响,并探讨其在新生血管形成中的作用及机制。方法将新生C57BL/6J小鼠、Fpr2^(-/-)小鼠分别诱导建立OIR模型后分为氧诱导组和Fpr2^(-/-)氧诱导组。正常对照组和Fpr2^(-/-)组小鼠正常环境饲养。收集各组小鼠视网膜组织并制作成冰冻切片,免疫荧光染色观察小鼠视网膜中波形蛋白(Vimentin)、神经胶质纤维酸性蛋白(GFAP)和离子化钙结合适配分子1(IBA1)变化;实时荧光定量PCR(RT-PCR)检测小鼠视网膜中炎症因子mRNA的表达;Western blot检测小鼠视网膜中细胞外调节蛋白激酶(ERK)和P38的蛋白磷酸化表达。免疫荧光染色法观察小鼠视网膜中新生血管和无灌注区情况。结果免疫荧光染色结果显示,与正常对照组相比,氧诱导组和Fpr2^(-/-)氧诱导组小鼠视网膜中Vimentin、GFAP、IBA1表达均明显增加,而与氧诱导组相比,Fpr2^(-/-)氧诱导组小鼠视网膜中Vimentin、GFAP、IBA1表达均降低(均为P<0.05)。RT-PCR检测结果显示,与Fpr2^(-/-)氧诱导组相比,氧诱导组小鼠视网膜中白细胞介素-1β(IL^(-1)β)、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)mRNA相对表达量均增加(均为P<0.05)。Western blot检测结果显示,与正常对照组相比,氧诱导组和Fpr2^(-/-)氧诱导组小鼠视网膜中p-ERK 1/2和p-P38蛋白相对表达量均增加,而与氧诱导组相比,Fpr2^(-/-)氧诱导组小鼠视网膜中p-ERK 1/2和p-P38蛋白相对表达量均减少(均为P<0.05)。与氧诱导组相比,Fpr2^(-/-)氧诱导组小鼠视网膜中新生血管形成和无灌注区面积均减少(均为P<0.05)。结论抑制Fpr2能够抑制OIR模型小鼠视网膜胶质细胞活化,下调炎症因子表达,减少视网膜新生血管形成和无灌注区面积,此过程中涉及ERK、P38信号通路的作用。
Objective To investigate the effect of the formyl peptide receptor(Fpr2)on the changes of glial cells in oxygen-induced retinopathy(OIR),and to explore its role and mechanism in neovascularization.Methods OIR model was induced in newborn C57BL/6J mice and Fpr2^(-/-)mice and they were divided into oxygen induced group and Fpr2^(-/-)oxygen induced group of OIR model.The mice in normal control group and Fpr2^(-/-)group were fed in normal environment.The retinal tissues of mice in each group were collected and made into frozen sections and stained with immunofluorescence staining.Then,the changes of glial cell markers including Vimentin,glial fibrillary acidic protein(GFAP)and ionized calcium binding aptamer 1(IBA1)were observed.The mRNA expressions of inflammatory factors in retinas in each group were detected by real time fluorescent quantitative PCR(RT-PCR).Western blot was used to detect the phosphorylation of extracellular regulated protein kinase(ERK)and P38 in retinas.The retinal neovascularization and non-perfusion area were observed by retinal flat-mount immunofluorescence staining.Results Immunofluorescence showed that the expression levels of Vimentin,GFAP and IBA1 in the oxygen induced group and Fpr2^(-/-)oxygen induced group significantly increased compared with those in the normal control group,while compared with the oxygen induced group,the expression levels of Vimentin,GFAP and IBA1 in the retina of mice in the Fpr2^(-/-)oxygen induced group decreased(all P<0.05).The RT-PCR results showed that,compared with the Fpr2^(-/-)oxygen induced group,the relative expression levels of interleukin-1β(IL^(-1)β),tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)mRNA in the retina of mice in the oxygen induced group increased(all P<0.05).The Western blot results showed that,compared with the normal control group,the relative expression levels of p-ERK1/2 and p-P38 protein in the retina of the oxygen induced group and Fpr2^(-/-)oxygen induced group increased,and compared with the oxygen induced group,the rel
作者
薛盛丁
陈辉
刘爽
朱蓉嵘
王诗逸
俞莹
XUE Shengding;CHEN Hui;LIU Shuang;ZHU Rongrong;WANG Shiyi;YU Ying(Department of Ophthalmology,Nanping First Hospital,Nanping 353000,Fujian Province,China;Department of Ophthalmology,Affiliated Hospital of Nantong University,Nantong 226001,Jiangsu Province,China)
出处
《眼科新进展》
CAS
北大核心
2021年第6期522-527,共6页
Recent Advances in Ophthalmology
基金
第65批中国博士后科学基金面上项目(编号2019M651927)
南通市科技计划项目(编号JCZ19081)。
