Protective effect of butyrate against ethanol-induced gastric ulcers in mice by promoting the anti-inflammatory, anti-oxidant and mucosal defense mechanisms

doi:10.1016/j.intimp.2015.11.018

International Immunopharmacology

Volume 30, January 2016, Pages 179-187

https://doi.org/10.1016/j.intimp.2015.11.018 Get rights and content

Highlights

•
This is the first demonstration that butyrate protects the gastric mucosa against ethanol-induced lesions.
•
Butyrate protected the gastric mucosa through anti-oxidative and anti-inflammatory mechanisms and through reinforcing the mucosal defense mechanisms.
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Butyrate, which is already in use in the clinic, could be used to treat gastric ulcer patients.

Abstract

Gastric ulcers (GUs) are a common type of peptic ulcer. Alcohol overdose is one of the main causes of GU, which is difficult to prevent. Although the protective effect of butyrate on inflammation-related diseases is well understood, its effect on GUs has not been reported. We investigated the protective effects of butyrate against ethanol-induced lesions to the gastric mucosa in mice and the underlying mechanisms. BALB/c mice were orally pretreated with butyrate for 30 min prior to the establishment of the GU model by challenge with absolute ethanol. Ethanol administration produced apparent mucosal injuries with morphological and histological damage, whereas butyrate pretreatment reduced the gastric mucosal injuries in a dose-dependent manner. Butyrate pretreatment also significantly ameliorated contents of malondialdehyde (MDA) and carbonyl proteins, and decreased levels of IL-1β, TNF-α and IL-6. The Western blot results consistently demonstrated that butyrate pretreatment attenuated the phosphorylation of NF-κB p65, p38 MAPK and ERKs in the gastric tissues. Additionally, gastric wall mucus (GWM), a parameter reflecting mucosal defense, was clearly increased by butyrate pretreatment. Butyrate pretreatment protects the gastric mucosa against ethanol-induced lesions by strengthening the mucosal defense and anti-oxidant and anti-inflammatory activities. As a necessary substance for the body, butyrate may be applied to the prevention and treatment of GUs.

Introduction

Butyric acid (BA), a short-chain fatty acid, is synthesized by the microflora in the colon and serves as the primary source of energy for colonocytes [1]. Butyrate is a stable derivative of BA, and has been verified to be effective against intestinal diseases. Several studies have reported that butyrate can be used to treat inflammatory bowel diseases (IBDs) [2], including acute radiation-induced proctitis [3], ulcerative colitis [4], [5] and Crohn's disease [6].

As an anti-cancer drug with inhibitory effects on cell proliferation in carcinoma cell lines [7], [8], butyrate has been clinically used to treat patients with a high risk of colon cancer [9]. More interestingly, butyrate was shown to have many potential applications in the extra-intestinal systems and organs [10]. For example, insulin resistance [11], anemia [12], [13], [14] and non-digestive tract cancers [15], [16] were improved by treatment with butyrate.

Although the adaptation diseases are rapidly expanding, butyrate has failed to attract attention for the treatment of injuries to the stomach. In fact, in 1959, butyrate was reported to increase the thickness of the gastric mucus layer in young calves [17]. There have been no subsequent studies on the protective effects of butyrate on the gastric mucosa.

As the main type of peptic ulcer, gastric ulcers (GUs) are the most common lesion of the gastric mucosa. Given that butyrate protects the gastric mucosa, butyrate may be a good complementary treatment, or even an alternative treatment, for GUs. Moreover, our published paper showed that Clostridium butyricum (C. butyricum), a BA-producing probiotic, prevented gastric mucosal injuries that were induced by various stimuli [18]. Therefore, it is reasonable to predict that BA is one of the factors leading to the protective effects of C. butyricum.

Various studies demonstrate that an imbalance of defense and offense is the basic mechanism underlying the development of GU. Reactive oxygen species (ROS) and excessive inflammatory reactions are two important factors in the pathogenesis of GUs. ROS not only directly damage the cell structures but also promote the production of proinflammatory factors, and proinflammatory cytokines, including IL-1β [19], TNF-α [20], [21] and IL-6 [22], have been shown to be key drivers of the development of GU.

In this study, the classical ethanol-induced GU mouse model was chosen for screening drugs with potentially protective effects against gastric mucosal lesions. We demonstrated that butyrate plays an important role protecting against ethanol-induced GU. Further investigations showed that butyrate exerts its biological effect by inhibiting the production of oxidative products and suppressing the production of proinflammatory factors which are induced by the activation of the NF-κB and MAP kinase signaling pathways, as well as by enhancing the production of gastric wall mucus (GWM). These results reveal a novel function for butyrate in protecting the gastric mucosa against lesions.

Section snippets

Reagents

Sodium butyrate was obtained from Sigma-Aldrich Co. (St. Louis, MO, USA). The detection kits for the carbonyl proteins and PAS staining were purchased from the Jiancheng Bioengineering Institute (Nanjing, China). The kits for malondialdehyde (MDA) and the bicinchoninic acid (BCA) protein assay were purchased from the Beyotime Institute of Biotechnology (Haimen, China). The ELISA kits for the mouse cytokines, including IL-1β, TNF-α and IL-6, were obtained from Westang Bio-Tech Co., Ltd.

Butyrate pretreatment alleviated the ethanol-induced macroscopic lesions to the gastric mucosa in mice

To observe the effects of butyrate on ethanol-induced GU, we performed macroscopic analyses on the gastric mucosal lesions. The pathological observations demonstrated that there were extensive hemorrhagic injuries, and even some shrinkage of the gastric wall on the surface of the gastric mucosa in mice after ethanol treatment (Fig. 1A). Interestingly, we found that butyrate pretreatment attenuated the mucosal lesions in a dose-dependent manner (Fig. 1A). Furthermore, the protective effects of

Discussion

GU has a high morbidity and a variety of etiologies, such as Helicobacter pylori (H. pylori) infection, excess free oxygen radicals, excessive acid, and non-steroidal anti-inflammatory drug (NSAID) sensitivity [28]. However, ethanol ingestion is one of the crucial contributors to gastric mucosal lesions [29], [30]. Although it is rare for GU to directly cause death, it has the potential to induce severe complications with a high mortality, such as gastric perforation, gastric bleeding and even

Author contributions

F. Wang and Y. Jiang conceived and designed the experiments. J. Liu, F. Wang, H. Luo, K. Li, and C. Li performed the experiments. F. Wang, H. Luo, and A. Liu analyzed the data. F. Wang and Y. Jiang wrote the paper.

Acknowledgments

This study was supported by grants from the National Natural Science Foundation of China (No. 81030055, No. 81372030 to Y.J.). We thank Dr. W.Z. Martini from the U.S. Army Institute of Surgical Research, Sam Houston, TX, USA for critically reading and editing this manuscript. The funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Protective effect of butyrate against ethanol-induced gastric ulcers in mice by promoting the anti-inflammatory, anti-oxidant and mucosal defense mechanisms

Highlights

Abstract

Introduction

Section snippets

Reagents

Butyrate pretreatment alleviated the ethanol-induced macroscopic lesions to the gastric mucosa in mice

Discussion

Author contributions

Acknowledgments

Curr. Probl. Pediatr. Adolesc. Health Care

Lancet

J. Nutr. Biochem.

Gastroenterology

Med. Hypotheses

J. Dairy Sci.

Biomed. Environ. Sci.

Anal. Biochem.

Methods Enzymol.

Gastroenterology

Phytomedicine

Trends Microbiol.

Int. J. Antimicrob. Agents

Pharmacol. Rep.

Chem. Biol. Interact.

Toxicol. Appl. Pharmacol.

J. Biol. Chem.

Toxicology

Alcohol

Chem. Biol. Interact.

Immunity

Potential beneficial effects of butyrate in intestinal and extraintestinal diseases

World J. Gastroenterol.

Oral butyrate for mildly to moderately active Crohn's disease

Aliment. Pharmacol. Ther.

The butyrate story: old wine in new bottles

Curr. Opin. Clin. Nutr. Metab. Care

A review of the potential mechanisms for the lowering of colorectal oncogenesis by butyrate

Br. J. Nutr.

Chemoprevention of colorectal cancer: systematic review and economic evaluation

Health Technol. Assess. (Winchester, England)

Butyrate and other short-chain fatty acids as modulators of immunity: what relevance for health

Curr. Opin. Clin. Nutr. Metab. Care

Butyrate improves insulin sensitivity and increases energy expenditure in mice

Diabetes