Fecal HBD-2 and Claudin-3 may be potential biomarkers to predict the deterioration of necrotizing enterocolitis: A prospective study

Abstract

Background and purpose: Necrotizing enterocolitis (NEC) is a critical gastrointestinal disease. We aim to explore the value of fecal human β-defensin 2 (HBD-2), Claudin-3, high-mobility group box-1 protein (HMGB-1), and resistin-like molecule β (Relmβ) as well as some laboratory metrics to predict the deterioration of NEC.

Methods: Infants diagnosed with NEC at Stage II were enrolled in our study. Those who progressed to Stage III were included in the Stage III group and the rest were included in the Stage II group. Clinical data and laboratory metrics of the infants were collected. Fecal samples of HBD2, HMGB-1, Claudin-3, and Relmβ collected during their enrollment were determined by using enzyme-linked immunosorbent assay (ELISA) kits. Student's t-test, the Mann-Whitney U test, the chi-square test, receiver operating characteristic (ROC), and logistic regression analysis were performed.

Results: Sixty infants diagnosed with NEC at Stage II were enrolled in our study, with 27 in the Stage III group (n = 27) and 33 in the Stage II group (n = 33). Although many of these NEC cases were late preterm and term infants, the infants in the Stage III group had a lower gestational age (P < 0.05). The incidence of gestational diabetes mellitus, peritonitis, intestinal adhesion, and sepsis was higher and more infants in the Stage III group underwent surgeries (P < 0.05). The levels of HBD-2 and Claudin-3 were higher and neutrophil count was lower in the Stage III group than in the Stage II Group, and the area under the curve (AUC) was 0.754, 0,755, and 0.666, respectively (P < 0.05). HBD-2 ≥ 1649.02 ng/g and Claudin-3 ≥ 2488.71 pg/g were included in the multivariate stepwise logistic regression analysis (P < 0.05), and the AUC of the model was 0.805 (95% CI: 0.688-0.922).

Conclusion: Fecal HBD-2 and Claudin-3 may be potential biomarkers to predict the deterioration of NEC from Stage II to Stage III.

Keywords: biomarker; claudin-3; deterioration; human β-defensin 2; necrotizing enterocolitis.

Figure 1

Comparison of laboratory metrics between two groups and ROC of neutrophil count. Neutrophil count in the Stage III group significantly decreased (P < 0.05). (A) Platelet count, serum sodium, pH, immature/total neutrophils, and mean artery pressure showed no differences between the two groups (P > 0.05). (B–F) The AUC of neutrophil count was 0.666 (P < 0.05) (G).

Figure 2

Comparison of fecal biomarkers between two groups and ROCs of HBD-2 and Claudin-3. The fecal concentrations of human β-defensin 2 (HBD-2) and Claudin-3 were significantly higher in the Stage III group than in the Stage II Group (P < 0.05) (A,B). High-mobility group box-1 protein (HMGB-1) and resistin-like molecule β (Relmβ) showed no differences between the two groups (P > 0.05) (C,D). The AUC of HBD-2 and Claudin-3 were 0.754 and 0,755, respectively (P < 0.05) (E,F).

Figure 3

ROC curve of fecal biomarkers. The AUC of human β-defensin 2 (HBD-2) ≥ 1649.02 ng/g, combined with Claudin-3 ≥ 2488.71 pg/g, to predict the deterioration of NEC was 0.805, and it showed a statistical significance (P < 0.05). ROC, receiver operating characteristic; AUC, area under the curve; NEC, necrotizing enterocolitis.