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      Probiotics VSL#3 Protect against Development of Visceral Pain in Murine Model of Irritable Bowel Syndrome

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          Abstract

          Background and Aims

          Irritable bowel syndrome (IBS) is linked to post-inflammatory and stress-correlated factors that cause changes in the perception of visceral events. Probiotic bacteria may be effective in treating IBS symptoms. Here, we have investigated whether early life administration of VSL#3, a mixture of 8 probiotic bacteria strains, protects against development of visceral hypersensitivity driven by neonatal maternal separation (NMS), a rat model of IBS.

          Methods

          Male NMS pups were treated orally with placebo or VSL#3 from days 3 to 60, while normal, not separated rats were used as controls. After 60 days from birth, perception of painful sensation induced by colorectal distension (CRD) was measured by assessing the abdominal withdrawal reflex (score 0–4). The colonic gene expression was assessed by using the Agilent Whole Rat Genome Oligo Microarrays platform and confirmed by real time PCR.

          Results

          NMS rats exhibited both hyperalgesia and allodynia when compared to control rats. VSL#3 had a potent analgesic effect on CRD-induced pain without changing the colorectal compliance. The microarray analysis demonstrated that NMS induces a robust change in the expression of subsets of genes (CCL2, NOS3, THP1, NTRK1, CCR2, BDRKRB1, IL-10, TNFRSF1B, TRPV4, CNR1 and OPRL1) involved in pain transmission and inflammation. TPH1, tryptophan hydroxylase 1, a validated target gene in IBS treatment, was markedly upregulated by NMS and this effect was reversed by VSL#3 intervention.

          Conclusions

          Early life administration of VSL#3 reduces visceral pain perception in a model of IBS and resets colonic expression of subsets of genes mediating pain and inflammation.

          Transcript profiling

          Accession number of repository for expression microarray data is GSE38942 ( http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE38942).

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          Most cited references34

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          Oral bacteriotherapy as maintenance treatment in patients with chronic pouchitis: a double-blind, placebo-controlled trial.

          Pouchitis is the major long-term complication after ileal pouch-anal anastomosis for ulcerative colitis. Most patients have relapsing disease, and no maintenance treatment study has been performed. We evaluated the efficacy of a probiotic preparation (VSL#3) containing 5 x 10(11) per gram of viable lyophilized bacteria of 4 strains of lactobacilli, 3 strains of bifidobacteria, and 1 strain of Streptococcus salivarius subsp. thermophilus compared with placebo in maintenance of remission of chronic pouchitis. Forty patients in clinical and endoscopic remission were randomized to receive either VSL#3, 6 g/day, or an identical placebo for 9 months. Patients were assessed clinically every month and endoscopically and histologically every 2 months or in the case of a relapse. Fecal samples were collected for stool culture before and after antibiotic treatment and each month during maintenance treatment. Three patients (15%) in the VSL#3 group had relapses within the 9-month follow-up period, compared with 20 (100%) in the placebo group (P < 0.001). Fecal concentration of lactobacilli, bifidobacteria, and S. thermophilus increased significantly from baseline levels only in the VSL#3-treated group (P < 0.01). These results suggest that oral administration of this new probiotic preparation is effective in preventing flare-ups of chronic pouchitis.
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            A new model of chronic visceral hypersensitivity in adult rats induced by colon irritation during postnatal development.

            The irritable bowel syndrome (IBS) is a common disorder characterized by abdominal pain in the setting of altered perception of viscerosensory stimuli. This so-called visceral hyperalgesia occurs in the absence of detectable organic disease in the peripheral organs and may cause normal or physiologic contractions to be perceived as painful. Although the pathogenesis of IBS remains speculative and is probably multifactorial, a prevailing paradigm is that transient noxious events lead to long-lasting sensitization of the neural pain circuit, despite complete resolution of the initiating event. Neonatal male Sprague-Dawley rats received either mechanical or chemical colonic irritation between postnatal days 8 and 21 and were tested when they became adults. The abdominal withdrawal reflex and the responses of viscerosensitive neurons were recorded during colon distention. Colon irritation in neonates, but not in adults, results in chronic visceral hypersensitivity, with characteristics of allodynia and hyperalgesia, associated with central neuronal sensitization in the absence of identifiable peripheral pathology. These results concur largely with observations in patients with IBS, providing a new animal model to study IBS and validating a neurogenic component of functional abdominal pain that encourages novel approaches to health care and research.
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              Neonatal maternal separation alters stress-induced responses to viscerosomatic nociceptive stimuli in rat.

              This study investigated the combined effect of neonatal maternal separation and acute psychological stress on pain responses in adult rats. Long-Evans dams and their male pups were reared under two conditions: 1) 180 min daily maternal separation (MS180) on postnatal days 2-14 or 2) no handling or separation (NH). At 2 mo of age, visceromotor responses to graded intensities of phasic colorectal distension (10-80 mmHg) at baseline as well as following acute 60 min water avoidance stress (WA) were significantly higher in MS180 rats. Both groups showed similar stress-induced visceral hyperalgesia in the presence of naloxone (20 mg/kg ip). MS180 rats had smaller stress-induced cutaneous analgesia in the tail-flick test compared with NH rats, with a residual naloxone-resistant component. MS180 rats showed an enhanced fecal pellet output following WA or exposure to a novel environment. These data suggest that early life events predispose adult Long-Evans rats to develop visceral hyperalgesia, reduced somatic analgesia, and increased colonic motility in response to an acute psychological stressor, mimicking the cardinal features of irritable bowel syndrome.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2013
                15 May 2013
                : 8
                : 5
                : e63893
                Affiliations
                [1 ]S.C. di Gastroenterologia ed Epatologia, Azienda Ospedaliera di Perugia, Perugia, Italy
                [2 ]Dipartimento di Medicina Clinica e Sperimentale, Università degli Studi di Perugia, Perugia, Italy
                University of California, Los Angeles, United States of America
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Conceived and designed the experiments: ED SF. Performed the experiments: SC AM BR. Analyzed the data: SC. Wrote the paper: ED BR SF.

                Article
                PONE-D-12-37671
                10.1371/journal.pone.0063893
                3655059
                23691109
                1d86d502-6d80-4915-8150-e65042b4c7e9
                Copyright @ 2013

                This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 1 December 2012
                : 7 April 2013
                Page count
                Pages: 15
                Funding
                This study was supported in part by CDS investments (Italy). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Agriculture
                Animal Management
                Animal Genetics
                Biology
                Anatomy and Physiology
                Digestive System
                Digestive Physiology
                Digestive Functions
                Digestive Regulation
                Computational Biology
                Molecular Genetics
                Gene Identification and Analysis
                Gene Regulation
                Gene Expression
                Microarrays
                Genetics
                Molecular Genetics
                Gene Identification and Analysis
                Animal Genetics
                Gene Expression
                Gene Function
                Gene Networks
                Genomics
                Genome Analysis Tools
                Genetic Networks
                Immunology
                Genetics of the Immune System
                Model Organisms
                Animal Models
                Rat
                Molecular Cell Biology
                Gene Expression
                Medicine
                Anatomy and Physiology
                Digestive System
                Digestive Physiology
                Digestive Functions
                Digestive Regulation
                Anesthesiology
                Pain Management
                Clinical Immunology
                Genetics of the Immune System
                Critical Care and Emergency Medicine
                Gastrointestinal Critical Care
                Diagnostic Medicine
                Pathology
                Clinical Pathology
                Molecular Genetics
                Epidemiology
                Genetic Epidemiology
                Gastroenterology and Hepatology
                Anal and Rectal Disorders
                Bacterial and Foodborne Illness
                Colon
                Gastrointestinal Infections
                Gastrointestinal Motility Disorders
                Inflammatory Bowel Disease
                Infectious Diseases
                Gastrointestinal Infections

                Uncategorized
                Uncategorized

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