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bioRxiv · 10.1101/2023.08.19.553983

Paneth cells disruption and intestinal dysbiosis contribute to the development of Hirschsprung-associated enterocolitis in a benzalkonium chloride-induced Hirschsprungs disease rat model

Budianto, I. R.·Kusmardi, K.·Maulana, A. M.·Arumugam, S.·Afrin, R.·Soetikno, V.
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Abstract

BackgroundHirschsprung-associated enterocolitis (HAEC) is a life-threatening complication of Hirschsprungs disease (HSCR). This study investigated the role of Paneth cells (PCs) and gut microbiota in HAEC development. MethodsMale 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. ResultsCompared 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 -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. ConclusionDisruption 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.

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Budianto, I. R., Kusmardi, K., Maulana, A. M., Arumugam, S., Afrin, R., Soetikno, V.. 2023-08-21. Paneth cells disruption and intestinal dysbiosis contribute to the development of Hirschsprung-associated enterocolitis in a benzalkonium chloride-induced Hirschsprungs disease rat model. https://doi.org/10.1101/2023.08.19.553983

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