A newly designed enteral formula containing whey peptides and fermented milk product protects mice against concanavalin A-induced hepatitis by suppressing overproduction of inflammatory cytokines

doi:10.1016/j.clnu.2011.10.012

Clinical Nutrition

Volume 31, Issue 2, April 2012, Pages 283-289

https://doi.org/10.1016/j.clnu.2011.10.012 Get rights and content

Summary

Background & aims

We previously reported that whey protein derived from cow milk suppressed inflammation in a variety of animal models. We developed a newly designed enteral formula using peptides prepared from whey protein and fermented milk product and investigated its ability to suppress inflammation in concanavalin A-induced hepatitis in mice.

Methods

C57BL/6 mice were fed a standard formula, AIN-93M, or enteral formula for 14 days, and then were intravenously administered concanavalin A. Inflammatory cytokines in plasma, liver, and spleen and markers of hepatic function in plasma were assessed at various time points. Livers were assessed for necrosis and apoptosis.

Results

After concanavalin A treatment, plasma aspartate aminotransaminase, alanine aminotransferase, TNF-α, IL-6, and IFN-γ levels were significantly lower in mice fed enteral formula than in those fed standard formula or AIN-93M. Liver TNF-α and IFN-γ, and spleen IL-6 and IFN-γ levels were lower in enteral formula-fed mice than in standard formula-fed mice 2 h after concanavalin A treatment. Necrosis and apoptosis were suppressed in the livers of enteral formula-fed mice.

Conclusions

The new enteral formula is a potent novel immune-modulating diet that prevents aggravation of local inflammation by modulating systemic cytokine levels.

Introduction

Inflammation is the primary host response to infection and injury. The reactions that take place in the course of inflammation are considered to be a series of events necessary to establish protective responses against infection and injury. However, persistence of the inflammatory response leads to serious tissue damage. Previous studies have shown that the persistence of inflammation is associated with a variety of disorders, many of which include inflammation as a major pathogenic cause.¹ Inflammatory cytokines, such as TNF-α and IL-6, are induced in liver diseases, including acute liver failure, viral and autoimmune hepatitis,2, 3, 4, 5, 6 postoperative liver and bowel ischemia,⁷ and sepsis.8, 9 Cytokine overproduction induces inflammation and is associated with the development and aggravation of hepatitis and with increased morbidity and mortality.9, 10

Whey proteins are one of the major classes of milk proteins. Although they are generally recognized as a source of quality amino acids, accumulating data from several studies have shown that they also regulate a variety of physiological events related to immunologic and neurologic functions.¹¹ We previously recognized that the whey proteins α-lactalbumin and β-lactoglobulin and their tryptic peptides have anti-inflammatory effects.12, 13, 14, 15, 16 First, we used a concanavalin A (Con A)-induced hepatitis model to evaluate the effects of whey proteins and their tryptic peptides and found that they prevent increases in plasma aspartate aminotransferase (AST) and alanine aminotransferase (ALT) via the regulation of TNF-α and IFN-γ production.12, 14 Second, the effects of whey protein and peptides were evaluated in a d-galactosamine-induced hepatitis model, a chemically induced hepatitis model.13, 15 It was found that whey proteins and peptides have hepatoprotective effects in which they prevent increases in plasma levels of hepatic function markers (i.e. AST, ALT, lactate dehydrogenase, and bilirubin) as well as hyaluronic acid, a fibrosis marker. Third, we investigated the effects of whey protein and peptides in a lipopolysaccharide-induced mouse sepsis model and found that they suppress the production of inflammatory cytokines such as TNF-α and IL-6.¹²

Based on these results, we used peptides to formulate a new enteral formula (EF) called MEIN. This formula has an anti-inflammatory function that is primarily attributed to the peptides. It also contains Lactobacillus bulgaricus- and Streptococcus thermophilus-fermented milk products, which are expected to modulate the immune reaction and improve the intestinal environment. In this report, we show that the new EF significantly suppressed the onset of inflammation in a Con A-induced hepatitis model.

Section snippets

Animals and diet

Six week-old male C57BL/6 Cr Slc mice (SLC, Hamamatsu, Japan), weighing 20–25 g, were used for the Con A-induced liver injury model. The mice were housed in an air-conditioned room at 21 ± 2 °C and 55 ± 15% humidity, with lights on from 7:00 to 19:00 h. Animals were handled according to the Guidelines for the Care and Use of Laboratory Animals of Meiji Co., Ltd. Mice were provided with drinking water and an experimental diet ad libitum.

The two different enteral diets, the new EF (MEIN; Meiji

Body weights, organ weights, and plasma biochemical analyses 24 h after Con A injection

Table 2 presents the body weights, organ weights, and plasma biochemical analyses. No differences were seen in the body weights between groups before and after Con A injection. The spleen weight per body weight (%) was significantly lower in SF-fed mice than in mice fed the other diets, and liver weight per body weight (%) was significantly lower in SF-fed than AIN-fed mice. Albumin and glucose levels were significantly lower in SF-fed than MEIN-fed mice. There were no significant differences

Discussion

A wide range of diseases are associated with acute or chronic inflammation.¹ Although survival of patients suffering from a variety of diseases has been extended with recent medical progress, long-term inflammation can be a cause of poor nutrition and reduced quality of life. Appropriate nutritional support options that can prevent or decrease inflammation and improve nutritional status are therefore warranted. Specifically, nutritional approaches that are able to either restrict the production

Conflict of interest statement

The authors have no conflicts of interest to declare.

Author agreement

All authors have made substantial contributions and final approval of the conceptions, drafting, and final version.

Acknowledgments

H K conceived the study design, conducted the studies, performed the data and statistical analyses, and drafted the manuscript. K O carried conducted the data analyses. H S and T Y participated in the study design, and helped in drafting of the manuscript. All authors read and approved the final manuscript. We would like to thank Hidekazu Tonouchi, Megumi Koganei, Kentaro Nakamura, Kentaro Ito and Akina Sasayama for their technical support.

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Original articleA newly designed enteral formula containing whey peptides and fermented milk product protects mice against concanavalin A-induced hepatitis by suppressing overproduction of inflammatory cytokines

Summary

Background & aims

Methods

Results

Conclusions

Introduction

Section snippets

Animals and diet

Body weights, organ weights, and plasma biochemical analyses 24 h after Con A injection

Discussion

Conflict of interest statement

Author agreement

Acknowledgments

Biomed Pharmacother

Cytokine

Hepatology

Am J Pathol

Gastroenterology

J Food Prot

Toxicology

Vitam Horm

Points of control in inflammation

Nature

Twelve potential fibrosis markers to differentiate mild liver fibrosis from cirrhosis in patients infected with chronic hepatitis C genotype 1

Eur J Clin Microbiol Infect Dis

Cytokines in viral hepatitis

Semin Liver Dis

TNF-alpha and growth hormone resistance in patients with chronic liver disease

J Interferon Cytokine Res

Fibromyalgia, hepatitis C infection, and the cytokine connection

Curr Pain Headache Rep

The immunopathogenesis of sepsis

Nature

Circulating cytokine/inhibitor profiles reshape the understanding of the SIRS/CARS continuum in sepsis and predict mortality

J Immunol

Original article
A newly designed enteral formula containing whey peptides and fermented milk product protects mice against concanavalin A-induced hepatitis by suppressing overproduction of inflammatory cytokines