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

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中文摘要：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pERKl/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pERKl/2的表达发挥作用。

外文标题：Silencing Nodal inhibits the biological behavior of retinal vascular endothelial cells under high glucose conditions

外文摘要：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(pERKl/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 pERKl/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 pERKl/2 expression.

外文关键词：

NodalRetinal vascular endothelial cellsNeovascularizationCell proliferationCell migrationLumen formation

作者：

曹靖靖、寇振宇、王晴、庄彤彤、东莉洁、王林妮

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作者单位：

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

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

基金：

天津市高等教育委员会科技发展基金项目天津市滨海新区卫生健康委员会科技项目天津市视网膜功能与疾病重点实验室开放项目天津市卫生健康科研项目天津市医学重点学科(专科)建设项目

项目编号：

2022ZD0572022BWKZ0032021tjswmm002TJWJ2023ZD002TJYXZDXK-037A

出版年：

2024

DOI：

10.3760/cma.j.cn511434-20230423-00181

中华眼底病杂志

中华医学会

中华眼底病杂志

CSTPCD北大核心

影响因子：0.928

ISSN：1005-1015

年,卷(期)：2024.40(2)

参考文献量18