First experience with the metaprebiotic Stimbifid plus used for eradication of Helicobacter pylori in patients with gastric ulcer

Abstract

Objective. To analyze the relationships between probiotic microorganisms and H. pylori KM-11 (RifR) and evaluate their effect on the structural organization of the pathogen and natural colonization resistance of the stomach both independently and with the metaprebiotic Stimbifid plus using bacteriological methods and electron microscopy. Our findings can be potentially used for effective eradication of H. pylori KM-11 (RifR) and treatment of gastric ulcer in volunteers during metaprebiotic therapy. Materials and methods. The following strains of microorganisms were used in this study: Lactobacillus plantarum 8P-A3, Bifidobacterium bifidum No 1, and Helicobacter pylori, isolated from a biopsy specimen of the pyloric antrum collected from a patient with gastritis. A rifampicin-resistant strain of H. pylori KM-11 (RifR) growing on a solid medium with rifampicin (160 μg∙mL–1) was obtained by spontaneous mutagenesis. H. pylori and H. pylori КМ-11 (RifR) were cultivated on hemin containing solid medium with special nutrients at 37°C in an anaerobic cultivation system. Microorganisms were identified by their morphological assessment and using kits for biochemical identification of bacteria. The relationships between probiotic bacteria and H. pylori KM-11 (RifR) were analyzed using the method of paired cultivation on solid and liquid media. The metaprebiotic Stimbifid plus was used in these experiments. Electron microscopy of all microorganisms was performed using a scanning electron microscope. Data analysis was conducted using the Kerber method modified by I.P. Ashmarin and A.A.Vorobyov. Results. Our in vitro experiments with paired cultivation of L. plantarum 8P-A3 and B. bifidum No.1 with H. pylori КМ-11 (RifR) on solid and liquid media containing Stimbifid plus showed that probiotic microorganisms were bioincompatible with H. pylori, i.e. there was an antagonism between a probiotic strain and a pathogenic microorganism. Stimbifid plus added to the cultivation medium acted as an anti-H. pylori agent; moreover, it promoted the restoration of colonization resistance and was a source of exclusive nutrients for probiotic bacteria. Bacteriological testing and electron microscopy demonstrated that the metabolites produced by L. plantarum 8P-A3 can damage the cell wall of H. pylori КМ-11 (RifR) during their co-cultivation in a liquid medium containig Stimbifid plus. This damage appeared as specific changes on the surface of the cell wall and resulted in the loss of viability. Oral administration of Stimbifid plus in six volunteers with gastric ulcer and concomitant severe dysbiosis (with 4 of them tested positive for H. pylori), ensured not only H. pylori eradication and treatment of gastric ulcer, but also confirmed the efficacy of an experimental dose of Stimbifid plus (3000 mg daily for 14 days). Conclusion. The results of our in vitro experiments with cocultivation of probiotic strains L. plantarum 8P-A3 and B. bifidum No 1 with H. pylori КМ-11 (RifR) on solid and liquid media containing Stimbifid plus, as well as experiments with oral administration of Stimbifid plus for H. pylori eradication and treatment of gastric ulcer, demonstrated a substantial therapeutic potential of this metaprebiotic, in particular as a therapy for chronic H. pylori infection and gastric ulcer scarring. Our current results and previous findings on the restoration of colonization resistance, gastric mucosa, and indigenous microbiota, as well as the data on the clearance of pathogenic bacteria in mammals, suggest that Stimbifid plus has a high eradication potential and can be used in clinical practice as a therapeutic agent for acute and chronic infections caused by H. pylori. Key words: Helicobacter pylori, microbiota, colonization resistance, gastric ulcer, eradication, metaprebiotic Stimbifid plus, volunteers