形觉剥夺性近视模型大鼠巩膜成纤维细胞中TGF-β1表达及Wnt/β-catenin信号通路的调控作用

doi:10.13481/j.1671-587x.20190420

摘要/Abstract

摘要： 目的：构建形觉剥夺性近视（FDM）大鼠模型，阐明FDM大鼠巩膜成纤维细胞中转化生长因子β1（TGF-β1）的表达及其与Wnt/β-连环蛋白（β-catenin）信号通路的关系。方法：40只大鼠随机分为对照组和FDM模型组，每组20只，FDM模型组大鼠建立FDM模型。测定大鼠眼轴长度；取大鼠眼球分离巩膜组织，采用Western blotting法和逆转录-聚合酶链反应（RT-PCR）法测定大鼠巩膜组织中TGF-β1蛋白和mNRA表达水平。分离培养对照组和FDM模型组大鼠巩膜成纤维细胞，对照组大鼠巩膜成纤维细胞作为对照组，FDM模型组大鼠巩膜成纤维细胞分为FDM组和FDM+Dickkopf相关蛋白1（DDK1）组（加入DDK1培养），采用Western blotting法和RT-PCR法测定各组大鼠巩膜成纤维细胞中TGF-β1、3型核糖核酸酶（DCL3）、结肠腺瘤样息肉蛋白（APC）、糖原合成酶激酶3β（GSK3β）、p21-GSK3β和β-catenin蛋白及mRNA表达水平。结果：与对照组比较，FDM组大鼠眼轴长度均明显增加（P<0.05），巩膜组织中TGF-β1蛋白和mRNA表达水平均明显降低（P<0.01）。对照组、FDM组和FDM+DDK1组大鼠巩膜成纤维细胞中GSK3β蛋白和mRNA表达水平比较差异无统计学意义（P>0.05），TGF-β1、DCL3、APC、p21-GSK3β和β-catenin蛋白及mRNA表达水平比较差异有统计学意义（P<0.01）；与对照组比较，FDM组大鼠巩膜成纤维细胞中TGF-β1、APC蛋白和mRNA表达水平降低（P<0.05），DCL3、p21-GSK3β、β-catenin蛋白和mRNA表达水平升高（P<0.05）；与FDM组比较，FDM+DDK1组大鼠巩膜成纤维细胞中TGF-β1、APC蛋白和mRNA表达水平升高（P<0.05），DCL3、p21-GSK3β、β-catenin蛋白和mRNA表达水平降低（P<0.05）。结论：FDM模型大鼠巩膜成纤维细胞中TGF-β1表达水平降低，其水平受Wnt/β-catenin信号通路调控。

关键词: 近视, 巩膜, 成纤维细胞, 转化生长因子β1, Wnt/β-连环蛋白, 大鼠,SD

Abstract: Objective:To construct the form deprivation myopia (FDM) rat models, and to elucidate the expression of transforming growth factor-β1 (TGF-β1) in scleral fibroblasts of the FDM rats and its relationship with Wnt/β-catenin signaling pathway. Methods:Forty rats were randomly divided into control group and FDM model group, with 20 rats in each group.The FDM rat model was established in FDM model group.The axial lengths of the rats were determined.The rat sclera tissue of eyeball was separated.The expression levels of TGF-β1 protein and mRNA in sclera tissue of the rats were determined by Western blotting and reverse transcription-polymerase chain reaction (RT-PCR) methods.The scleral fibroblasts of rats were isolated and cultured.The fibroblasts in the sclera of the rats in control group were used as the control group.The fibroblasts in FDM model group were divided into FDM group and FDM+Dickkopf related protein 1 (DDK1) group (added to DDK1 to culture).The expression levels of TGF-β1, Dicer-like 3 (DCL3), colon adenomatous polyp protein (APC), glycogen synthase kinase 3β (GSK3β), p21-GSK3β, and β-catenin protein and mRNA in scleral fibroblasts were determined by Western blotting and RT-PCR methods. Results:Compared with control group,the axial length of the rats in FDM group was increased (P<0.01);the expression levels of TGF-β1 protein and mRNA in sclera tissue were significantly decreased(P<0.01).There were no significant differences in the expression levels of GSK3β protein and mRNA in scleral fibroblasts between control group,FDM group and FDM+DDK1 group (P>0.05);but there were significant differences in the expression levels of TGF-β1,DCL3,APC,p21-GSK3β,and β-catenin protein and mRNA in the scleral fibroblasts(P<0.01).Compared with control group, the expression levels of TGF-β1 and APC protein and mRNA in scleral fibroblasts of the rats in FDM group were decreased(P<0.01), and the expression levels of DCL3, p21-GSK3β, and β-catenin protein and mRNA were increased(P<0.05).Compared with FDM group, the expression levels of TGF-β1 and APC protein and mRNA in the scleral fibroblasts of the rats in FDM+DDK1 group were increased(P<0.01), and the expression levels of DCL3, p21-GSK3β, and β-catenin protein and mRNA were decreased(P<0.01). Conclusion:The expression level of TGF-β1 in the scleral fibroblasts of the FDM model rats is decreased, and its level is regulated by the Wnt/β-catenin signaling pathway.

Key words: myopia, sclera, fibroblasts, transforming growth factor-β1, Wnt/β catenin, rats,SD

中图分类号:

R778.11

张新, 巨朝娟, 张剑, 赵燕. 形觉剥夺性近视模型大鼠巩膜成纤维细胞中TGF-β1表达及Wnt/β-catenin信号通路的调控作用[J]. 吉林大学学报(医学版), 2019, 45(04): 861-866.

ZHANG Xin, JU Zhaojuan, ZHANG Jian, ZHAO Yan. Expression of TGF-β1 in scleral fibroblasts of rats with form deprivation myopia and regulation of Wnt/β-catenin signaling pathway[J]. Journal of Jilin University(Medicine Edition), 2019, 45(04): 861-866.

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