Protective effects of hydrogen-rich saline on necrotizing enterocolitis in neonatal rats

J Pediatr Surg. 2013 Aug;48(8):1697-706. doi: 10.1016/j.jpedsurg.2012.11.038.


Purpose: The aim of this study was to test the hypothesis that hydrogen-rich saline (HRS) might have protective effects on the development of necrotizing enterocolitis (NEC) in a neonatal rat model.

Methods: NEC was induced in male newborn Sprague-Dawley rats by formula feeding, exposure to asphyxia and cold stress. Sixty-four rat pups were divided randomly into four groups: C+NS (n=11), C+H2 (n=11), NEC+NS (n=20), and NEC+H2 (n=22). Rats in the former two groups were mother-fed. Pups received intra-peritoneal injection of HRS (10 ml/kg, 10 min before asphyxia stress twice a day) or the same dose of normal saline. Rats were monitored until 96 h after birth. Body weight, histological NEC score, survival time, malondialdehyde, antioxidant capacity, inflammatory mediators, and mucosal integrity were assessed.

Results: HRS treatment maintained the body weight, reduced the incidence of NEC from 85% (17/20) to 54.5% (12/22), increased the survival rate from 25% (5/20) to 68.2% (15/22), and attenuated the severity of NEC. In addition, HRS inhibited the mRNA expression of pro-inflammatory mediators (inducible nitric oxide synthase, tumor necrosis factor-α, and interleukin-6), down-regulated lipid peroxidation, enhanced total antioxidant capacity, and prevented the increase of diamine oxidase in serum. However, no significant influence of HRS on the interleukin-10 mRNA expression was observed.

Conclusions: HRS showed beneficial effects on neonatal rats with NEC via decreasing oxidative stress, increasing antioxidant capacity, suppressing inflammation, and preserving mucosal integrity.

Keywords: Hydrogen-rich saline; Inflammation; Necrotizing enterocolitis; Oxidative stress.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amine Oxidase (Copper-Containing) / blood
  • Animals
  • Animals, Newborn
  • Antioxidants / analysis
  • Asphyxia / complications
  • Body Weight
  • Cold Temperature / adverse effects
  • Drug Evaluation, Preclinical
  • Enterocolitis, Necrotizing / drug therapy
  • Enterocolitis, Necrotizing / prevention & control*
  • Enzyme Induction / drug effects
  • Gene Expression Regulation / drug effects
  • Hydrogen / administration & dosage
  • Hydrogen / therapeutic use*
  • Ileum / drug effects
  • Ileum / pathology
  • Infant Food / adverse effects
  • Injections, Intraperitoneal
  • Interleukin-10 / biosynthesis
  • Interleukin-10 / genetics
  • Interleukin-6 / biosynthesis
  • Interleukin-6 / genetics
  • Intestinal Mucosa / drug effects
  • Intestinal Mucosa / pathology
  • Lipid Peroxidation / drug effects
  • Male
  • Nitric Oxide Synthase Type II / biosynthesis
  • Nitric Oxide Synthase Type II / genetics
  • Oxidative Stress / drug effects
  • Random Allocation
  • Rats
  • Rats, Sprague-Dawley
  • Sodium Chloride / administration & dosage
  • Sodium Chloride / therapeutic use
  • Tumor Necrosis Factor-alpha / biosynthesis
  • Tumor Necrosis Factor-alpha / genetics


  • Antioxidants
  • Interleukin-6
  • Tumor Necrosis Factor-alpha
  • Interleukin-10
  • Sodium Chloride
  • Hydrogen
  • Nitric Oxide Synthase Type II
  • Nos2 protein, rat
  • Amine Oxidase (Copper-Containing)