Abstract
Background Hirschsprung-associated enterocolitis (HAEC) is a life-threatening complication of Hirschsprung’s disease (HSCR). This study investigated the role of Paneth cells (PCs) and gut microbiota in HAEC development.
Methods Male Sprague-Dawley rats with HSCR were established by exposure of 0.1% (n = 30) benzalkonium chloride (BAC) to rectosigmoid serosa and sacrificed at 1-, 3-, 5-, 8-, and 12-weeks postintervention. The sham group was included and sacrificed on Week 12. Hematoxylin-Eosin staining was conducted to count the number of ganglionic cells and analyze the degree of enterocolitis. Intestinal barrier function was assessed for the ratio of anti-peripherin, occludin and acetylcholinesterase (AChE)/butyrylcholinesterase (BChE). PCs antimicrobial peptide (AMP) was evaluated by cryptdins, secretory Phospholipase A2, and lysozyme levels by qRT-PCR, respectively. 16S rRNA high throughput sequencing on faecal samples was used to analyze the changes in intestinal microbiota diversity in each group.
Results Compared with sham groups, 0.1% BAC group rats had fewer ganglion cells after 1-week postintervention. Occludin and peripherin were decreased, and AChE/BChE ratio was increased, respectively. Sigmoid colon tissues from BAC-treated rats showed increased α-defensins positive PCs on Week 5 postintervention. Conversely, PCs-produced AMP tended to decrease from Week 5 to Week 12. Rats in the sham group demonstrated increased Lactobacillus and decreased Bacteroides, while rats in the 0.1% BAC exhibited reciprocal changes. Enterocolitis occurred from Week 1 postintervention onwards.
Conclusion Disruption of PCs in the Week 5 postintervention and dysbiosis exacerbate the occurrence of HAEC. This research sheds new light on the cellular mechanisms of HAEC development.
Methods Male Sprague-Dawley rats with HSCR were established by exposure of 0.1% (n = 30) benzalkonium chloride (BAC) to rectosigmoid serosa and sacrificed at 1-, 3-, 5-, 8-, and 12-weeks postintervention. The sham group was included and sacrificed on Week 12. Hematoxylin-Eosin staining was conducted to count the number of ganglionic cells and analyze the degree of enterocolitis. Intestinal barrier function was assessed for the ratio of anti-peripherin, occludin and acetylcholinesterase (AChE)/butyrylcholinesterase (BChE). PCs antimicrobial peptide (AMP) was evaluated by cryptdins, secretory Phospholipase A2, and lysozyme levels by qRT-PCR, respectively. 16S rRNA high throughput sequencing on faecal samples was used to analyze the changes in intestinal microbiota diversity in each group.
Results Compared with sham groups, 0.1% BAC group rats had fewer ganglion cells after 1-week postintervention. Occludin and peripherin were decreased, and AChE/BChE ratio was increased, respectively. Sigmoid colon tissues from BAC-treated rats showed increased α-defensins positive PCs on Week 5 postintervention. Conversely, PCs-produced AMP tended to decrease from Week 5 to Week 12. Rats in the sham group demonstrated increased Lactobacillus and decreased Bacteroides, while rats in the 0.1% BAC exhibited reciprocal changes. Enterocolitis occurred from Week 1 postintervention onwards.
Conclusion Disruption of PCs in the Week 5 postintervention and dysbiosis exacerbate the occurrence of HAEC. This research sheds new light on the cellular mechanisms of HAEC development.
| Original language | English |
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| DOIs | |
| Publication status | Published - 21 Aug 2023 |
Keywords
- Hirschsprung disease
- aganglionosis
- inflammation
- antimicrobial peptide
- enterocolitis