Analysis of gut microbiota and serum I-FABP and PRDX1 levels in children with enterocolitis after congenital megacolon surgery

Abstract

BACKGROUND

Hirschsprung’s disease (HD) is a congenital intestinal malformation defined by intestinal aganglionosis. Persistent spasm of the aganglionic segment is commonly followed by fecal retention and impaction, with consequent proximal colonic dilation and wall thickening. The only established curative treatment is surgery; nevertheless, postoperative Hirschsprung-associated enterocolitis (HAEC) occurs in approximately 30% of affected children, compromising physiological recovery and causing potentially life-threatening morbidity. Postoperative HAEC has been linked to impaired intestinal barrier function, gut microbial dysbiosis and immune dysregulation; however, the underlying mechanisms remain incompletely resolved. Prophylactic probiotic administration has been reported to reduce HAEC incidence and severity in some cohorts, whereas other studies have found minimal or no preventive benefit. Accordingly, both the contribution of gut microbiota structure to postoperative HAEC and the clinical utility of prophylactic probiotics in children undergoing HD surgery require further investigation.

AIM

To characterize the gut microbiota and serum levels of intestinal fatty acid-binding protein (I-FABP)/peroxiredoxin 1 (PRDX1) in children with HAEC following surgery, analyze their correlation, and evaluate the predictive value of preoperative gut microbiota and serum levels of I-FABP/PRDX1 in the development of postoperative HAEC.

METHODS

This retrospective study included 334 children with HD who underwent surgery at our hospital between March 2022 and March 2025. Patients were categorized into two groups based on the occurrence of HAEC within 3 months postoperatively: The HAEC group (n = 109) and the non-HAEC group (n = 225). The two groups were compared in terms of general characteristics, preoperative gut microbiota α-diversity indices, phylum and genus abundances, and serum levels of C-reactive protein (CRP), tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), interleukin-8 (IL-8), I-FABP, and PRDX1. Correlations between gut microbiota phylum/genus abundance and serum inflammatory markers were also analyzed. Logistic regression analysis was used to identify the risk factors for postoperative HAEC, and a nomogram prediction model was developed. The predictive performance of the model was validated internally using bootstrap resampling.

RESULTS

The HAEC group was characterized by a longer aganglionic segment and a higher incidence of preoperative enteritis and postoperative intestinal obstruction than the non-HAEC group (P < 0.05). No significant between-group differences were observed in gut microbiota α-diversity, as assessed by the Chao1, Shannon and Simpson indices (P > 0.05). By contrast, relative abundances of Proteobacteria, Bacteroidetes, Verrucomicrobia, Streptococcus and Bacteroides were increased, whereas those of Actinobacteria, Tenericutes, Bifidobacterium and Lactobacillus were decreased, in the HAEC group compared with the non-HAEC group (P < 0.05). Preoperative serum concentrations of CRP, TNF-α, IL-6, IL-8, I-FABP and PRDX1 were also significantly elevated in the HAEC group (P < 0.05). Correlation analysis demonstrated negative associations between these serum markers and the abundance of beneficial bacteria, alongside positive associations with pathogenic bacterial abundance. Multivariable regression analysis identified serum CRP, I-FABP and PRDX1 concentrations as independent risk factors for postoperative HAEC, whereas Bifidobacterium abundance emerged as an independent protective factor. On this basis, a nomogram prediction model was constructed incorporating preoperative abundances of Verrucomicrobia, Bacteroides and Bifidobacterium, together with serum CRP, I-FABP and PRDX1 levels. The model showed excellent discriminative performance, with an area under the receiver operating characteristic curve of 0.951 (95%CI: 0.927-0.974) and good calibration (Hosmer-Lemeshow χ² = 3.573, P = 0.893).

CONCLUSION

Children who develop HAEC following HD surgery exhibit a characteristic preoperative profile of reduced beneficial bacteria, increased pathogenic bacteria, and elevated levels of CRP, TNF-α, IL-6, IL-8, I-FABP, and PRDX1. Preoperative levels of these serum markers were significantly correlated with gut microbiota abundance. Furthermore, the risk of postoperative HAEC was strongly associated with preoperative gut microbiota structure as well as serum I-FABP and PRDX1 levels.

Keywords: Hirschsprung’s disease; Hirschsprung-associated enterocolitis; Gut microbiota; Intestinal fatty acid-binding protein; Peroxiredoxin 1

Core Tip: This retrospective study examined the characteristics of, and correlations between, preoperative gut microbiota and serum intestinal fatty acid-binding protein (I-FABP) and peroxiredoxin 1 (PRDX1) levels in children who developed Hirschsprung-associated enterocolitis (HAEC) after Hirschsprung’s disease (HD) surgery, with the aim of informing risk assessment and preventive intervention strategies. Preliminary analyses indicated that children with postoperative HAEC exhibited decreased abundance of beneficial bacteria, increased abundances of pathogenic bacteria and elevated preoperative levels of C-reactive protein (CRP), tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), interleukin-8 (IL-8), I-FABP and PRDX1. Preoperative levels of CRP, TNF-α, IL-6, IL-8, I-FABP and PRDX1 were significantly correlated with gut microbiota abundance, and the risk of HAEC after HD was strongly associated with preoperative gut microbiota structure and with I-FABP and PRDX1 levels.