先天性肛门直肠畸形大鼠后肠发育中Fibronectin与Laminin β1的表达及意义

Expression and Significance of Fibronectin and Laminin β1 During the Developmental Hindgut in Rats with Anorectal Malformations

该文采用全反式维甲酸(ATRA)构建SD大鼠肛门直肠畸形模型(n=32),探究FN1(fibronectin 1)与LAMB1(laminin β1)在大鼠胚胎后肠发育过程的表达及意义。对照组与模型组均于E11.5~E16.5、E18.5、E20.5剖宫取胎,记录胎鼠身长、体质量、尾长、大体形态。qRT-PCR、Western blot、免疫组织化学检测FN1与LAMB1 mRNA及蛋白在对照组及模型组胚胎后肠发育中的表达。对照组生长发育指标在各个时间点均优于模型组,差异具有统计学意义(P<0.05)。FN1与LAMB1在胚胎后肠发育中呈连续动态表达。对照组中,qRT-PCR、Western blot、免疫组织化学结果显示二者表达高峰为E15.5,主要均匀分布于泄殖腔上皮黏膜层和基底膜。在模型组中,二者mRNA和蛋白表达水平在E11.5~E16.5均显著上调,表达高峰较对照组滞后出现在E16.5,差异具有统计学意义(P<0.05),免疫组织化学结果提示,二者在泄殖腔中呈现出高表达和分布不均匀的改变。与对照组相比,ATRA诱导的肛门直肠畸形模型组胚胎生长发育滞后,后肠发育中FN1与LAMB1表达上调且峰值延迟,可能与先天性肛门直肠畸形发生有关。

The aim of this paper is to investigate the expression and significance of FN1(fibronectin 1)and LAMB1(laminin β1)in the development of hindgut in embryos with anorectal malformations(n=32),which induced by ATRA(all-trans retinoic acid).Embryos were collected at E11.5-E16.5,E18.5,and E20.5 in control group and model group,with the length,weight,tail length,and gross morphology of the fetuses recorded.qRTPCR,Western blot and immunohistochemistry were used to detect the expression of FN1 and LAMB1 in the hindgut development of control group and model group.In control group,the data of length,weight,tail length were superior to the model group at each developmental time point,and the differences were statistically significant(P<0.05).FN1 and LAMB1 expressed continuously during the hindgut development with regular changing in both group.In control group,the results of qRT-PCR,Western blot and immunohistochemistry showed that the expression levels of the two genes reached the peak at E15.5 and most of the two proteins homogeneously distributed in the epithelial mucosa and basement membrane of the cloaca.In model group,the expression levels of both genes were significantly up-regulated in E11.5-E16.5,and the peaks appeared at E16.5 compared with control group,the differences were statistically significant(P<0.05).Immunohistochemistry results showed that both expressed highly with uneven distribution in the cloaca.Compared with control group,the embryo development of model group which induced by ATRA was delayed,and the expression of FN1 and LAMB1 were up-regulated with the peaks delayed during the hindgut development,which might be related to the form of anorectal malformations.