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.
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