Elsevier

Nutrition

Volume 33, January 2017, Pages 96-104
Nutrition

Basic nutritional investigation
Administration of probiotic mixture DM#1 ameliorated 5-fluorouracil–induced intestinal mucositis and dysbiosis in rats

https://doi.org/10.1016/j.nut.2016.05.003Get rights and content

Highlights

  • Administration of probiotic mixture DM#1 ameliorated 5-fluorouracil–induced mucositis and dysbiosis in rats.

  • Administration of DM#1 promoted reestablishment of intestinal microbial homeostasis.

  • Administration of DM#1 also altered the toll-like receptor 2 signaling pathway.

Abstract

Objective

The use of probiotics to alleviate chemotherapy-induced intestinal mucositis is supported by clinical consensus. However, no studies to date, to our knowledge, have systematically analyzed the effects of a probiotic mixture on chemotherapy-induced mucositis or assessed changes in the intestinal microbiota after probiotic treatment. The aim of this study was to report the effects of a probiotic mixture, DM#1, on intestinal mucositis and dysbiosis of rats treated with 5-fluorouracil (5-FU).

Methods

Twenty-eight male Sprague Dawley rats weighing 180 to 220 g were randomly divided into four groups: control, 5-FU, probiotic high (PH), and probiotic low (PL). Except for the control group, all other groups received intraperitoneal injections of 5-FU for 5 d, and the PH and PL groups received DM#1 intragastrically (1 × 109 or 1 × 108 colony-forming units/kg, respectively) for 8 d. One day after the last administration, rats were sacrificed and the ilea were removed for histopathologic assessment and evaluation of permeability, myeloperoxidase activity, levels of cytokines (interleukin [IL]-4, IL-6, tumor necrosis factor [TNF]-α), and mRNA of toll-like receptors (TLR; TLR2, TLR4, and TLR9). Additionally, intestinal microbiota profiles were analyzed by polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis and quantitative real-time PCR.

Results

Treatment with DM#1 ameliorated 5-FU–induced intestinal mucosal injury in rats, possibly by reducing proinflammatory cytokine levels and neutrophil infiltration. The increased intestinal permeability caused by 5-FU was ameliorated. These results were closely associated with the reestablishment of intestinal microbial homeostasis and alteration of the TLR2/TLR4 signaling pathway.

Conclusions

Administration of the probiotic mixture DM#1 ameliorated 5-FU–induced intestinal mucositis and dysbiosis in rats.

Introduction

More than 50% of patients receiving the chemotherapy drug 5-fluorouracil (5-FU) develop either oral or intestinal mucositis [1]. Injury of the intestinal mucosal barrier is one of the most debilitating side effects of 5-FU treatment and is associated with pain, bacteremia, and malnutrition [2]. These complications result from reduced enterocyte proliferation and migration and increased cell apoptosis, which combine to disrupt normal intestinal barrier function [3]. Destruction of the intestinal mucosa leads to reduced nutrient absorption and increased vulnerability to infection [4].

In addition to inducing intestinal mucositis, chemotherapeutics also have detrimental effects on the composition of the intestinal microbiota. A shift in composition from predominantly gram-positive to predominantly gram-negative bacteria was observed following administration of 5-FU in Lewis rats [5]. A regimen of 5-FU and irinotecan elevated Clostridium cluster XI and Enterobacteriaceae [6]. Because commensal bacteria play pivotal roles in both the innate and adaptive immune systems of the host [7], intestinal dysbiosis contributes greatly to the development of chemotherapy-induced mucositis and diarrhea [8]. Therefore, normalization of intestinal homeostasis could be an appropriate strategy to improve the status of patients receiving chemotherapy.

In recent years, the use of probiotics to alleviate damage to intestinal mucosa has been supported by clinical consensus [9]. Both live probiotic strains and factors derived from probiotic culture supernatants have been tested in animal models to examine their effects on 5-FU–induced intestinal mucositis. Streptococcus thermophilus TH-4 improved the mitotic count, decreased crypt fission, and reduced histologic deficits caused by 5-FU [10]. Administration of probiotic factors derived from Escherichia coli Nissle 1917 and Lactobacillus fermentum BR11 partially protected the intestine from 5-FU–induced mucositis [11]. Furthermore, treatment with Lactobacillus acidophilus improved the inflammatory and functional aspects of 5-FU–induced intestinal mucositis [12]. In contrast, Lactobacillus rhamnosus GG and Bifidobacterium lactis BB12 did not alleviate mucositis caused by 5-FU [13]. However, to our knowledge, no studies have systematically analyzed the effects of a probiotic mixture on 5-FU–induced mucositis or assessed changes in the intestinal microbiota after probiotic intervention.

In the present study, we investigated the effects of a probiotic mixture, DM#1, on intestinal mucositis and dysbiosis of rats treated with 5-FU. DM#1 includes four previously isolated probiotic strains: Bifidobacterium breve DM8310, L. acidophilus DM8302, Lactobacillus casei DM8121, and S. thermophilus DM8309. We also investigated changes in intestinal barrier function after treatment with 5-FU and/or DM#1 and evaluated signaling pathways that may be involved in the regulation of intestinal barrier function. The results of this study will provide a valuable complementary reference for clinical treatment as well as theoretical support for the development of targeted probiotic therapy.

Section snippets

Bacterial culture

The probiotic preparation is a probiotic mixture of four strains, namely B. breve DM8310, L. acidophilus DM8302, L. casei DM8121, and S. thermophilus DM8309. They were originally isolated from Chinese fermented food and were deposited in the Culture Collection of Dalian Medical University (DMCC), Dalian, China. To identify the strains, 16 S rDNA were amplified by polymerase chain reaction (PCR) and submitted to sequencing. The partial sequences of 16 S rDNA of the four strains were deposited in

Effects of DM#1 on mortality, body weight, and metabolism parameters of 5-FU–treated rats

The experiment design is shown in Figure 1A, during the 8-d experiment, two rats died after 5-FU treatment in the model group (5-FU group) on days 3 and 4, respectively (Fig. 1B). In the low-dose (5-FU + PL) and high-dose (5-FU + PH) probiotics-treated groups, one rat from each group died on day 4. No control group rats died.

The average BW of rats in the control group increased from 202.29 ± 12.88 g to 239.14 ± 15.65 g throughout the study period, the changing of body weight (ΔBW) increased by

Discussion

The normal microbiota dwelling in the human intestinal tract antagonizes the growth of pathogens, promotes immune function, affects nutritional status, plays a role in detoxification, and has antiaging effects [21]. Therefore, the intestinal microbiota is important to the health of the host. Patients receiving cytotoxic therapy exhibit marked changes in the structure and composition of the intestinal microbiota. These changes most frequently include decreases in Bifidobacterium spp., Clostridium

Conclusions

Results from the present study indicated that, for treatment with 5-FU induced dysbiosis in the rat intestine, both microbial diversity and the populations of individual microbial groups were affected. Furthermore, this shifting of the intestinal microbiota may contribute greatly to the destruction of the mucosal barrier in rats by influencing the TLR2 signaling pathway. Treatment with DM#1 ameliorated 5-FU–induced intestinal mucosal injury in rats, possibly by reducing inflammatory factors

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    This work was supported by the Key Project of the National Twelfth-Five Year Research Program of China (2012 BAI35 B02), the National High-Technology Research and Development Program of China (863 Program, 2014 AA022200), the Research Fund for the Doctoral Program of Higher Education, China (RFDP, 20132105120012), and the Nature Science Foundation of Liaoning Province, China (2015020262). YT and YW contributed equally to this work.

    The authors have no conflicts of interest to declare.

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