Hydrogen-rich water protects against inflammatory bowel disease in mice by inhibiting endoplasmic reticulum stress and promoting heme oxygenase-1 expression

World J Gastroenterol. 2017 Feb 28;23(8):1375-1386. doi: 10.3748/wjg.v23.i8.1375.

Abstract

Aim: To investigate the therapeutic effect of hydrogen-rich water (HRW) on inflammatory bowel disease (IBD) and to explore the potential mechanisms involved.

Methods: Male mice were randomly divided into the following four groups: control group, in which the mice received equivalent volumes of normal saline (NS) intraperitoneally (ip); dextran sulfate sodium (DSS) group, in which the mice received NS ip (5 mL/kg body weight, twice per day at 8 am and 5 pm) for 7 consecutive days after IBD modeling; DSS + HRW group, in which the mice received HRW (in the same volume as the NS treatment) for 7 consecutive days after IBD modeling; and DSS + HRW + ZnPP group, in which the mice received HRW (in the same volume as the NS treatment) and ZnPP [a heme oxygenase-1 (HO-1) inhibitor, 25 mg/kg] for 7 consecutive days after IBD modeling. IBD was induced by feeding DSS to the mice, and blood and colon tissues were collected on the 7th d after IBD modeling to determine clinical symptoms, colonic inflammation and the potential mechanisms involved.

Results: The DSS + HRW group exhibited significantly attenuated weight loss and a lower extent of disease activity index compared with the DSS group on the 7th d (P < 0.05). HRW exerted protective effects against colon shortening and colonic wall thickening in contrast to the DSS group (P < 0.05). The histological study demonstrated milder inflammation in the DSS + HRW group, which was similar to normal inflammatory levels, and the macroscopic and microcosmic damage scores were lower in this group than in the DSS group (P < 0.05). The oxidative stress parameters, including MDA and MPO in the colon, were significantly decreased in the DSS + HRW group compared with the DSS group (P < 0.05). Simultaneously, the protective indicators, superoxide dismutase and glutathione, were markedly increased with the use of HRW. Inflammatory factors were assessed, and the results showed that the DSS + HRW group exhibited significantly reduced levels of TNF-α, IL-6 and IL-1β compared with the DSS group (P < 0.05). In addition, the pivotal proteins involved in endoplasmic reticulum (ER) stress, including p-eIF2α, ATF4, XBP1s and CHOP, were dramatically reduced after HRW treatment in contrast to the control group (P < 0.05). Furthermore, HRW treatment markedly up-regulated HO-1 expression, and the use of ZnPP obviously reversed the protective role of HRW. In the DSS + HRW + ZnPP group, colon shortening and colonic wall thickening were significantly aggravated, and the macroscopic damage scores were similar to those of the DSS + HRW group (P < 0.05). The histological study also showed more serious colonic damage that was similar to the DSS group.

Conclusion: HRW has a significant therapeutic potential in IBD by inhibiting inflammatory factors, oxidative stress and ER stress and by up-regulating HO-1 expression.

Keywords: Endoplasmic reticulum stress; Heme oxygenase-1; Hydrogen; Inflammatory bowel disease; Oxidative stress.

MeSH terms

  • Activating Transcription Factor 4 / metabolism
  • Animals
  • Colon / pathology
  • Endoplasmic Reticulum Stress*
  • Eukaryotic Initiation Factor-2 / metabolism
  • Gene Expression Regulation, Enzymologic
  • Heme Oxygenase-1 / blood*
  • Hydrogen / chemistry*
  • Inflammatory Bowel Diseases / drug therapy
  • Inflammatory Bowel Diseases / prevention & control*
  • Interleukin-1beta / blood
  • Interleukin-6 / blood
  • Male
  • Membrane Proteins / blood*
  • Mice
  • Mice, Inbred C57BL
  • Oxidative Stress
  • Random Allocation
  • Time Factors
  • Transcription Factor CHOP / metabolism
  • Tumor Necrosis Factor-alpha / blood
  • Water / chemistry*
  • X-Box Binding Protein 1 / metabolism

Substances

  • Atf4 protein, mouse
  • Ddit3 protein, mouse
  • Eukaryotic Initiation Factor-2
  • Interleukin-1beta
  • Interleukin-6
  • Membrane Proteins
  • Tumor Necrosis Factor-alpha
  • X-Box Binding Protein 1
  • Xbp1 protein, mouse
  • Water
  • Activating Transcription Factor 4
  • Transcription Factor CHOP
  • Hydrogen
  • Heme Oxygenase-1
  • Hmox1 protein, mouse