Dishevelled2和Vangl2表达与过量维甲酸致昆明小鼠神经管畸形的相关性

Relationship between the expression of Dishevelled2 and Vangl2 and the embryonic neural tube defects induced by excessive retinoic acid in Kunming mice

目的探讨Dishevelled2和Vangl2蛋白与过量维甲酸致昆明小鼠神经管畸形发生的关系。方法50只昆明孕小鼠随机分为对照组和实验组。孕7.75d时,实验组一次性胃饲30mg/kg致畸量维甲酸(RA),对照组胃饲等量溶剂,服药后4h、18h、42h、66h及90h分别取胚胎,常规石蜡切片,采用原位杂交和免疫组织化学技术,分别检测胚胎神经管组织中Dishevelled2蛋白和Vangl2mRNA及蛋白的表达水平变化。结果两种蛋白均存在于对照组及实验组不同发育阶段的小鼠神经管上皮,且两者的表达变化趋势有所不同。与对照组相比,实验组Dishevelled2蛋白在RA处理18h、42h时表达水平明显降低(P≤0.05),66h时表达升高(P≤0.05),90h时两组表达水平无明显差异;Vangl2mRNA在灌服RA4h、18h时表达水平明显低于对照组(P≤0.05),42h时两组表达水平无明显差异,66h时表达显著高于对照组(P≤0.05)。Vangl2蛋白在RA处理18h、42h时表达显著降低(P≤0.05),66h时两组表达水平无明显差异,90h时表达显著高于对照组(P≤0.05)。结论过量RA可能通过调节Dishevelled2和Vangl2表达干扰正常胚胎神经管的闭合过程。

Objective To explore the relationship between the expression of Dishevelled2 and Vangl2 and the embryonic neural tube defects （NTDs） induced by all-trans retinoic acid （RA）in Kunming mouse. Methods Fifty pregnant mice were randomly divided into control and RA-treated groups.RA-treated mice were fed with 30mg/kg RA dissolved with peanut oil on embryo 7.75 days, while the mice of control group were administrated with an equal volume of peanut oil on the same time. Then all the embryos were sampled from pregnant mice at the 4th, 18th, 42nd, 66th and 90th hour after treatment. In situ hybridization and immunohistochemical staining technique were used to detect the expression of Dishevelled2 and Vangl2 in embryonic neural tube. Results The two proteins both existed in the epithelial tissue of the mouse embryonic neural tube and displayed different expression modes at various developmental stages.Compared with the control group, the RA treated group showed a significant decrease （P≤0.05） at the 18th and 42nd hour and a significant increase （P≤0.05） at the 66th hour in Dishevelled2 protein after maternal treatment, and no significant difference was found at the 90th hour. Compared with the control group, the Vangl2 mRNA expression in the RA treated group displayed a significant decrease （P≤0.05） at the 4th and 18th hour and a significant increase （P≤0.05） at the 66th hour after RA treatment, and no difference was found at the 42nd hour. Compared with the control group, the expression of Vangl2 protein in the RA treated group decreased （P≤0.05） at the 18th and 42nd hour, and increased （P≤0.05） at the 90th hour after RA treatment, no difference was found at the 66th hour. Conclusion Excessive RA may interfere with the normal embryonic neural tube closure by regulating the expression of Dishevelled2 and Vangl2.