Notch信号通路对体外培养小鼠角膜缘上皮细胞黏液素5AC表达的影响  

Effect of Notch Signaling Pathway on Mucin 5AC Expression in Corneal Limbal Epithelial Cells of Mice

作　　者：郑恬[1,2] 熊林杰[1] 胡义珍[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院眼科 [2]武汉大学中南医院眼科

出　　处：《华中科技大学学报（医学版）》2014年第3期281-284,共4页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

摘　　要：目的通过体外细胞分化模型探讨Notch信号通路对角膜缘上皮细胞分化过程中黏液素5AC(MUC5AC)表达的影响及其临床意义。方法 Western blot检测体外细胞培养24和48h时Notch1、Notch2及Notch3蛋白在小鼠角膜缘上皮细胞中的表达情况;以小鼠体外正常培养角膜缘上皮细胞作为对照组,靶向转染Notch3siRNA的细胞作为实验组,培养24、48和72h后运用Western blot和Real-time PCR检测MUC5AC蛋白及mRNA的表达水平;RT-PCR检测转谷氨酰胺酶1mRNA表达强度变化。结果 Western blot结果显示Notch3蛋白在角膜缘上皮细胞的表达最高,其次是Notch2,Notch1表达不明显。与对照组相比,实验组MUC5AC的蛋白合成及mRNA表达显著下降,其蛋白合成于24、48和72h分别下降了(64.3±35.0)%、(39.5±11.0)%和(41.3±8.3)%;mRNA分别下降了(42.9±14.2)%、(45.0±18.0)%和(46.4±21.0)%;转谷氨酰胺酶1的基因表达明显增强,24、48和72h分别增加了(75.2±11.3)%、(89.7±15.5)%和(101.0±25.0)%。结论 Notch信号通路可影响MUC5AC基因及蛋白合成,对Notch信号的调节可能是治疗黏液素分泌不足型眼表疾病的一个新途径。Objective To investigate the effect of the Notch signaling pathway on the expression of mucin 5AC in the corneal limbal epithelial cells via the in-vitro cell differentiation model. Methods Western blot was employed to detect the expressions of Notch1, Notch2 and Notch3 in the mouse corneal limbal epithelial cells cultured for 24 and 48 h in vitro. Western blot and real-time PCR were used to measure the expressions of mucin SAC protein and mRNA in corneal limbal epithelial cells 24, 48 and 72 h after transfection with Notch3 siRNA,with the normal corneal epithelial cells serving as control group. The mRNA expression of transglutaminase 1 was detected by RT-PCR. Results Western blot showed that the protein expression of Notch3 was highest in corneal limbal epithelial cells, followed by that of Notch2. The expression of Notchl was unnoticeable. Compared with the control group, the protein expression of mucin 5 AC was decreased by （ 64.3± 35.0） %, （39.5±11.0） % and （41.3 ± 8.3） % 24,48 and 72 h after transfection with siRNA,while the mRNA expression of mucin 5AC was decreased by （42.9 ±14.2） %, （45.0±18.0） % and （46.4±21.0） %, respectively. The expression of transglutaminase 1 was enhanced by （75.2 ± 11.3）%,（89. 7±15.5）% and （101.0±25.0）% 24,48 and 72 h after transfection with siRNA. Conclusion Notch signaling pathway may affect the expression of mucin 5AC protein and mRNA. Regulation of Notch signaling may be a new therapy strategy for mucin-deficient ocular disease.

关 键 词：NOTCH信号 角膜缘上皮细胞分化 黏液素5AC 转谷氨酰胺酶1 

分 类 号：R772.2[医药卫生—眼科]