Helicobacter pylori, Protected from Antibiotics and Stresses Inside Candida albicans Vacuoles, Cause Gastritis in Mice

Abstract

Due to (i) the simultaneous presence of Helicobacter pylori (ulcer-induced bacteria) and Candida albicans in the stomach and (ii) the possibility of prokaryotic–eukaryotic endosymbiosis (intravacuolar H. pylori in the yeast cells) under stresses, we tested this symbiosis in vitro and in vivo. To that end, intravacuolar H. pylori were induced by the co-incubation of C. albicans with H. pylori under several stresses (acidic pH, non-H. pylori-enrichment media, and aerobic environments); the results were detectable by direct microscopy (wet mount) and real-time polymerase chain reaction (PCR). Indeed, intravacuolar H. pylori were predominant under all stresses, especially the lower pH level (pH 2–3). Interestingly, the H. pylori (an amoxicillin-sensitive strain) inside C. albicans were protected from the antibiotic (amoxicillin), while extracellular H. pylori were neutralizable, as indicated by the culture. In parallel, the oral administration of intravacuolar H. pylori in mice caused H. pylori colonization in the stomach resulting in gastritis, as indicated by gastric histopathology and tissue cytokines, similar to the administration of free H. pylori (extra-Candida bacteria). In conclusion, Candida protected H. pylori from stresses and antibiotics, and the intravacuolar H. pylori were able to be released from the yeast cells, causing gastric inflammation with neutrophil accumulations.

The Regulatory Roles of Ezh2 in Response to Lipopolysaccharide (LPS) in Macrophages and Mice with Conditional Ezh2 Deletion with LysM-Cre System

“…Notably, the supernatant of the stimulated cells in all groups was gently removed, and washed with DMEM, before the re-administration of LPS or DMEM as mentioned in previous publications [ 24 , 102 , 103 ]. Supernatant cytokines (TNF-α, IL-6, and IL-10) were evaluated by ELISA (Invitrogen, Carlsbad, CA, USA) and the gene expression was evaluated by quantitative real-time polymerase chain reaction (PCR) as previously described [ 104 , 105 , 106 , 107 ]. Briefly, the RNA was extracted from the cells with TRIzol Reagent (Invitrogen, Carlsbad, CA, USA) together with RNeasy Mini Kit (Qiagen, Hilden, Germany) as 1 mg of total RNA was used for cDNA synthesis with iScript reverse transcription supermix (Bio-Rad, Hercules, CA, USA).…”

High Fructose Causes More Prominent Liver Steatohepatitis with Leaky Gut Similar to High Glucose Administration in Mice and Attenuation by Lactiplantibacillus plantarum dfa1

“…Cells at 1 × 10 6 cells/well were then incubated with an additional 25 mM/well of glucose or fructose with or without 100 µg/mL of lipopolysaccharide (LPS) from E. coli O26: B6 (Sigma-Aldrich, St. Louis, MO, USA) before determining supernatant cytokines (Quantikine Immunoassay; R & D Systems, Minneapolis, MN, USA). For enterocytes, the expression of occludin (intestinal tight junction) and the nuclear factor kappa B ( NF-κB ) in relative to β-actin (a housekeeping gene) was carried out according to the 2 −ΔΔCp method [ 60 ]. The primers were as follows; occludin , forward 5′-CCAATGTCGAGGAGTGGG-3′, reverse 5′-CGCTGCTGTAACGAGGCT-3′; NF-κB , forward 5′-AGCACAGATACCACCAAGACC-3′, reverse 3′-GGGCACGATTGTCAAAGAT-5′; β-actin forward 5′-CCTGGCACCCAGCACAAT-3′, reverse 5′-GCCGATCCACACGGAGTACT-3′.…”

Lacticaseibacillus rhamnosus dfa1 Attenuate Cecal Ligation-Induced Systemic Inflammation through the Interference in Gut Dysbiosis, Leaky Gut, and Enterocytic Cell Energy

“…After that, this cell-free supernatant (0.5 mL) was concentrated by speed vacuum drying at 40 °C for 3 h (Savant Instruments, Farmingdale, NY, USA) and stored at −20 °C until use [ 101 ]. Then, the LCM pellets, resuspended in DMEM (5% LCM) or DMEM alone (Media), in an equal volume were incubated in the Caco-2 cells with starvation before the sample collection for supernatant cytokines by ELISA (Invitrogen) or gene expression with quantitative real-time polymerase chain reaction (PCR) following previous publications [ 102 , 103 , 104 ].…”