High intestinal vascular permeability in a murine model for Hirschsprung’s disease: implications for postoperative Hirschsprung-associated enterocolitis

• Published in: Pediatric surgery international Vol. 39; no. 1; p. 15
• Main Authors: Suda, Kazuto, Yamada, Shunsuke, Miyahara, Katsumi, Fujiwara, Naho, Kosaka, Seitaro, Abe, Kumpei, Seo, Shogo, Nakamura, Shinji, Lane, Geoffrey J., Yamataka, Atsuyuki
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 30.11.2022; Springer Nature B.V
• Subjects: Angiogenesis; Animals; Antibodies; Capillary Permeability; Colon; Disease; Disease Models, Animal; Enterocolitis - etiology; Evans Blue; Hirschsprung Disease - complications; Laboratories; Medicine; Medicine & Public Health; Mice; Original; Original Article; Pediatric Surgery; Pediatrics; Permeability; Polymerase chain reaction; Postoperative period; Rodents; Small intestine; Surgery; Vascular endothelial growth factor; Vascular Endothelial Growth Factor A - genetics; Vascular Endothelial Growth Factor B; Endothelin receptor-B; Vascular integrity; Hirschsprung associated enterocolitis; Vascular permeability; Vascular endothelial growth factor; Hirschsprung’s disease
• ISSN: 1437-9813; 0179-0358; 1437-9813
• DOI: 10.1007/s00383-022-05308-7

Purpose Intestinal vascular permeability (VP) in a murine model for Hirschsprung’s disease (HD) and postoperative Hirschsprung-associated enterocolitis (HAEC) were investigated. Methods Intestinal VP was determined using a Miles assay using 1% Evans blue injected into a superficial temporal vein of newborn endothelin receptor-B KO HD model (KO) and syngeneic wild-type (WT) mice ( n  = 5, respectively). Extravasated Evans blue in normoganglionic ileum (Ng-I), normoganglionic proximal colon (Ng-PC) and aganglionic distal colon (Ag-DC) was quantified by absorbance at 620 nm. Quantitative polymerase chain reaction (qPCR) for Vascular Endothelial Growth Factor A (VEGF-A), VEGF-B, CDH5, SELE and CD31, and immunofluorescence for CD31 were performed. Results VP was significantly higher in Ng-I, Ng-PC, and Ag-DC from KO than WT ( p  < 0.01, p  < 0.05, and p  < 0.05, respectively). qPCR demonstrated upregulated VEGF-A in Ng-I and Ag-DC, VEGF-B in Ng-I, and SELE in Ng-I and Ng-PC ( p  < 0.05, p  < 0.05, p  < 0.05, p  < 0.01 and p  < 0.05, respectively), and downregulated CDH5 in Ng-I and Ng-PC from KO ( p  < 0.05, respectively). Expression of CD31 mRNA in Ng-I and Ag-DC from KO was significantly higher on qPCR ( p  < 0.05) but differences on immunofluorescence were not significant. Conclusions VP may be etiologic for postoperative HAEC throughout the intestinal tract even after excision of aganglionic bowel.