LncRNA NEAT1调控miR-505-3p/VEGFA对碱烧伤大鼠角膜新生血管的影响  

Effect of long non-coding RNA nuclear paraspeckle assembly transcript 1 on corneal neovascularization in alkali-burned rats by regulating the microRNA-505-3p/vascular endothelial growth factor A

作　　者：程新潮 曹瑾 杨洁 吕旭东 CHENG Xinchao;CAO Jin;YANG Jie;L?Xudong(Department of Ophthalmology,Xianning Central Hospital,Xianning 437100,Hubei Province,China)

机构地区：[1]咸宁市中心医院眼科,湖北省咸宁市437100

出　　处：《眼科新进展》2023年第11期863-868,共6页Recent Advances in Ophthalmology

基　　金：湖北省卫生健康委员会面上项目(编号:WJ2019M097);2021年度咸宁市中心医院院级科研项目(编号:2021XYB017)。

摘　　要：目的 探讨长链非编码RNA(LncRNA)核旁斑组装转录本1(NEAT1)调控微小RNA-505-3p(miR-505-3p)/血管内皮生长因子A(VEGFA)轴对碱烧伤大鼠角膜新生血管生成的影响及其作用机制。方法 144只SD大鼠随机分为对照组、模型组、sh-NC组、sh-NEAT1组、sh-NEAT1+antagomir-NC组和sh-NEAT1+miR-505-3p antagomir组,每组24只。除对照组外,其余组大鼠均建立碱烧伤模型。观察大鼠角膜新生血管情况,并计算角膜新生血管面积;实时荧光定量PCR(qRT-PCR)检测大鼠角膜中LncRNA NEAT1、miR-505-3p、VEGFA mRNA表达;苏木素-伊红(HE)染色观察大鼠角膜病理形态学改变;免疫组织化学染色、Western blot检测大鼠角膜中VEGFA、CD31蛋白表达;双荧光素酶报告基因实验验证miR-505-3p与LncRNA NEAT1、VEGFA的靶向关系。结果 与对照组比较,模型组大鼠角膜新生血管面积增加,LncRNA NEAT1、VEGFA mRNA相对表达水平以及VEGFA、CD31蛋白表达水平均升高,miR-505-3p相对表达水平降低(均为P<0.05),角膜组织存在炎症细胞浸润、上皮细胞缺损等病理学损伤;与sh-NC组比较,sh-NEAT1组大鼠角膜新生血管面积减少,LncRNA NEAT1、VEGFA mRNA相对表达水平以及VEGFA、CD31蛋白表达水平均降低,miR-505-3p相对表达水平升高(均为P<0.05),角膜组织病理学损伤有所改善;与sh-NEAT1+antagomir-NC组比较,sh-NEAT1+miR-505-3p antagomir组大鼠角膜新生血管面积增加,VEGFA mRNA相对表达水平以及VEGFA、CD31蛋白表达水平均升高,miR-505-3p相对表达水平降低(均为P<0.05),角膜组织病理学损伤加重。经验证,大鼠角膜上皮细胞中miR-505-3p与LncRNA NEAT1、VEGFA均存在靶向关系。结论 干扰LncRNA NEAT1可能通过靶向上调miR-505-3p并下调VEGFA表达抑制碱烧伤大鼠角膜新生血管生成。Objective To investigate the effect of long non-coding RNA(LncRNA)nuclear paraspeckle assembly transcript 1(NEAT1)on corneal neovascularization in alkali-burned rats by regulating the microRNA-505-3p(miR-505-3p)/vascular endothelial growth factor A(VEGFA)axis.Methods A total of 144 SD rats were randomly divided into the control group,model group,sh-NC group,sh-NEAT1 group,sh-NEAT1+antagomir-NC group,and sh-NEAT1+miR-505-3p antagomir group,with 24 rats in each group.Except for the control group,alkali-burn rat models were established in the rest groups.Corneal neovascularization was observed,and its area was measured.The mRNA levels of LncRNA NEAT1,miR-505-3p and VEGFA in the rat cornea were detected by quantitative real-time polymerase chain reaction.The pathomorphological changes in the rat cornea were observed by hematoxylin-eosin staining.The expression levels of VEGFA and CD31 proteins in the rat cornea were measured by immunohistochemical staining and Western blot.The targeting relationship between miR-505-3p and LncRNA NEAT1,VEGFA was verified by the dual luciferase reporter gene assay.Results Compared with the control group,the corneal neovascularization area and the relative expression levels of LncRNA NEAT1 and VEGFA mRNAs as well as VEGFA and CD31 proteins in the model group increased,while the relative expression level of miR-505-3p decreased(all P<0.05),and pathological damages such as inflammatory cell infiltration and epithelial cell defects existed in corneal tissues.Compared with the sh-NC group,the corneal neovascularization area and the relative expression levels of LncRNA NEAT1 and VEGFA mRNAs as well as VEGFA and CD31 proteins in the sh-NEAT1 group decreased,while the relative expression level of miR-505-3p increased(all P<0.05),and the corneal histopathological damages improved.Compared with the sh-NEAT1+antagomir-NC group,the corneal neovascularization area and the relative expression levels of VEGFA mRNA,VEGFA protein and CD31 protein in the sh-NEAT1+miR-505-3p antagomir group increased,whi

关 键 词：碱烧伤 角膜新生血管 长链非编码RNA核旁斑组装转录本1 微小RNA-505-3p 血管内皮生长因子A 

分 类 号：R772.2[医药卫生—眼科]