沉默Nodal抑制高糖条件下培养的视网膜血管内皮细胞的生物学行为  

Silencing Nodal inhibits the biological behavior of retinal vascular endothelial cells under high glucose conditions

作　　者：曹靖靖 寇振宇 王晴 庄彤彤 东莉洁 王林妮 Cao Jingjing;Kou Zhenyu;Wang Qing;Zhuang Tongtong;Dong Lijie;Wang Linni(Tianjin Key Laboratory of Retinal Functions and Diseases,Tianjin Branch of National Clinical Research Center for Ocular Disease,Eye Institute and School of Optometry,Tianjin Medical University Eye Hospital,Tianjin 300384,China)

机构地区：[1]天津医科大学眼科医院、眼视光学院、眼科研究所,国家眼耳鼻喉疾病临床医学研究中心天津市分中心,天津市视网膜功能与疾病重点实验室,天津300384

出　　处：《中华眼底病杂志》2024年第2期136-141,共6页Chinese Journal of Ocular Fundus Diseases

基　　金：天津市高等教育委员会科技发展基金项目(2022ZD057);天津市滨海新区卫生健康委员会科技项目(2022BWKZ003);天津市视网膜功能与疾病重点实验室开放项目(2021tjswmm002);天津市卫生健康科研项目(TJWJ2023ZD002);天津市医学重点学科(专科)建设项目(TJYXZDXK-037A)。

摘　　要：目的观察高糖状态下Nodal对猴视网膜血管内皮细胞(RF/6A细胞)生物学行为的影响。方法将RF/6A细胞分为正常组、甘露醇组、高糖组、高糖联合非特异小干扰RNA处理组(HG+NC组)、高糖联合小干扰Nodal处理组(HG+siNodal组)。蛋白质免疫印迹法、实时荧光定量聚合酶链反应观察细胞内Nodal蛋白、mRNA相对表达量;噻唑蓝比色法检测Nodal对RF/6A细胞增殖的影响;细胞划痕实验检测Nodal对RF/6A细胞迁移能力的影响;Matrigel体外三维成型法检测Nodal对RF/6A细胞管腔形成的影响;蛋白质免疫印迹法检测RF/6A细胞内磷酸化细胞外调节蛋白激酶1/2(pERK1/2)蛋白相对表达量。多组间比较采用单因素方差分析。结果与HG+NC组比较,HG+siNodal组细胞内Nodal蛋白(F=33.469)、mRNA相对表达量(F=38.191)显著下降,细胞增殖能力(F=28.548)、细胞迁移能力(F=24.182)显著降低,细胞管腔形成数量显著减少(F=52.643),差异均有统计学意义(P<0.05)。与HG+NC组比较,HG+siNodal组细胞内pERK1/2蛋白相对表达量显著降低,差异有统计学意义(F=44.462,P<0.01)。结论沉默Nodal表达可抑制高糖诱导的RF/6A细胞增殖、迁移及成管能力;其可能通过抑制pERK1/2的表达发挥作用。Objective To observe the effect of Nodal on the biological behavior of retinal vascular endothelial cells(RF/6A cells)in monkeys with high glucose.Methods RF/6A cells were divided into normal group,mannitol group,high glucose group,high glucose combined with non-specific small interfering RNA treatment group(HG+NC group),high glucose combined with small interfering Nodal treatment group(HG+siNodal group).The transfection efficiency of siNodal was observed by real-time fluorescence quantitative PCR and western blot protein immunoblotting.The effect of Nodal on the proliferation of RF/6A cells was detected by thiazole blue colorimetry.The effect of Nodal on migration ability of RF/6A cells was detected by cell scratch assay.The effect of Nodal on the formation of RF/6A cell lumen was measured by Matrigel three-dimensional in vitro.The expression of extracellular signal phosphorylated regulated kinase 1/2(pERK1/2)in RF/6A cells was detected by western blot protein immunoblotting.One-way analysis of variance was used to compare groups.Results Compared with HG+NC group,Nodal protein(F=33.469)and mRNA relative expression levels(F=38.191)in HG+siNodal group were significantly decreased,cell proliferation was significantly decreased(F=28.548),and cell migration ability was significantly decreased(F=24.182).The number of cell lumen formation was significantly decreased(F=52.643),and the differences were statistically significant(P<0.05).Compared with HG+NC group,the relative expression of pERK1/2 protein in HG+siNodal group was significantly decreased,and the difference was statistically significant(F=44.462,P<0.01).Conclusions Silencing Nodal expression can inhibit proliferation,migration and tube formation of RF/6A cells induced by high glucose.It may act by inhibiting pERK1/2 expression.

关 键 词：NODAL 视网膜血管内皮细胞 新生血管 细胞增殖 细胞迁移 管腔形成 

分 类 号：R774.1[医药卫生—眼科] R587.2[医药卫生—临床医学]