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

WU Fang^1,2, GU Chengchao², BI Yang², GUO Zhenhua¹, WANG Yi^1*

(¹Department of Neonatal Gastrointestinal Surgery, Ministry of Education Key Laboratory of Child Development and Disorders, National Clinical Research Center for Child Health and Disorders (Chongqing), China International Science and Technology Cooperation base of Child development and Critical Disorders, Children’s Hospital of Chongqing Medical University, Chongqing 400014, China; ²Chongqing Key Laboratory of Pediatrics, Chongqing Engineering Research Center of Stem Cell Therapy, Chongqing 400014, China)

Abstract:

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.

CSTR: 32200.14.cjcb.2020.01.0002