Microbiota-gut-brain axis: Interaction of gut microbes and their metabolites with host epithelial barriers.

The gastrointestinal barrier and the blood brain barrier represent an important line of defense to protect the underlying structures against harmful external stimuli. These host barriers are composed of epithelial and endothelial cells interconnected by tight junction proteins along with several other supporting structures. Disruption in host barrier structures has therefore been implicated in various diseases of the gastrointestinal tract and the central nervous system. While there are several factors that influence host barrier, recently there is an increasing appreciation of the role of gut microbiota and their metabolites in regulating barrier integrity. In the current issue of Neurogastroenterology and Motility, Marungruang et al. describe the effect of gastrointestinal barrier maturation on gut microbiota and the blood brain barrier adding to the growing evidence of microbiota-barrier interactions. In this mini-review I will discuss the effect of gut microbiota on host epithelial barriers and its implications for diseases associated with disrupted gut-brain axis.

Impaired propulsive motility in the distal but not proximal colon of BK channel ß1-subunit knockout mice.

BACKGROUND: Large-conductance Ca(2+) -activated K(+) (BK) channels regulate smooth muscle tone. The BK channel ß1-subunit increases Ca(2+) sensitivity of the α-subunit in smooth muscle. We studied ß1-subunit knockout (KO) mice to determine if gastrointestinal (GI) motility was altered. METHODS: Colonic and intestinal longitudinal muscle reactivity to bethanechol and colonic migrating motor complexes (CMMCs) were measured in vitro. Gastric emptying and small intestinal transit were measured in vivo. Colonic motility was assessed in vivo by measuring fecal output and glass bead expulsion time. Myoelectric activity of distal colon smooth muscle was measured in vitro using intracellular microelectrodes. KEY RESULTS: Bethanechol-induced contractions were larger in the distal colon of ß1-subunit KO compared to wild type (WT) mice; there were no differences in bethanechol reactivity in the duodenum, ileum, or proximal colon of WT vsß1-subunit KO mice. There were more retrogradely propagated CMMCs in the distal colon of ß1-subunit KO compared to WT mice. Gastrointestinal transit was unaffected by ß1-subunit KO. Fecal output was decreased and glass bead expulsion times were increased in ß1-subunit KO mice. Membrane potential of distal colon smooth muscle cells from ß1-subunit KO mice was depolarized with higher action potential frequency compared to WT mice. Paxilline (BK channel blocker) depolarized smooth muscle cells and increased action potential frequency in WT distal colon. CONCLUSIONS & INFERENCES: BK channels play a prominent role in smooth muscle function only in the distal colon of mice. Defects in smooth muscle BK channel function disrupt colonic motility causing constipation.

5-HT(3) and 5-HT(4) receptors contribute to the anti-motility effects of Garcinia buchananii bark extract in the guinea-pig distal colon.

BACKGROUND: Garcinia buchananii bark extract is an anti-motility diarrhea remedy. We investigated whether G. buchananii bark extract has components that reduce gastrointestinal peristaltic activity via 5-HT(3) and 5-HT(4) receptors. METHODS: Aqueous G. buchananii extract was separated into fractions using preparative thin layer chromatography (PTLC), and major chemical components were identified using standard tests. The anti-motility effects of the extract and its fractions (PTLC1-5) were studied through pellet propulsion assays using isolated guinea-pig distal colons. KEY RESULTS: Anti-motility (PTLC1 & PTLC5) and pro-motility (PTLC2) fractions were isolated from the extract. Flavonoids, steroids, alkaloids, tannins, and phenols were identified in the extract and PTLC1&5. The potency of the extract applied via the mucosal surface was reduced by 5-HT, 5-HT(3) receptor agonist RS-56812, 5-HT(4) receptor agonists cisapride and CJ-033466, 5-HT(3) receptor antagonist granisetron, and 5-HT(4) receptor antagonist GR-113808. The anti-motility effects of the aqueous extract and PTLC1&5 when applied serosally were reversed by RS-56812, cisapride, and CJ-033466. The 5-HT(3) receptor antagonists, granisetron and ondansetron, reduced the effects of the extract to an extent and completely reversed the anti-motility effects of PTLC1&5. GR-113808 inhibited the actions of the extract during the initial 10 min, but enhanced the extracts' anti-motility effects after 15 min. GR-113808 augmented the anti-motility activities of PTLC1 and PTLC5 by 30%. CONCLUSIONS & INFERENCES: These results indicate that the anti-motility effects of G. buchananii aqueous extract are potentially mediated by compounds that affect 5-HT(3) and 5-HT(4) receptors. Identification and characterization of the bioactive compounds within G. buchananii could lead to the discovery of new non-opiate anti-diarrhea formulations.

The traditional antidiarrheal remedy, Garcinia buchananii stem bark extract, inhibits propulsive motility and fast synaptic potentials in the guinea pig distal colon.

BACKGROUND: Garcinia buchananii bark extract is a traditional African remedy for diarrhea, dysentery, abdominal discomfort, and pain. We investigated the mechanisms and efficacy of this extract using the guinea pig distal colon model of gastrointestinal motility. METHODS: Stem bark was collected from G. buchananii trees in their natural habitat of Karagwe, Tanzania. Bark was sun dried and ground into fine powder, and suspended in Krebs to obtain an aqueous extract. Isolated guinea pig distal colon was used to determine the effect of the G. buchananii bark extract on fecal pellet propulsion. Intracellular recording was used to evaluate the extract action on evoked fast excitatory postsynaptic potentials (fEPSPs) in S-neurons of the myenteric plexus. KEY RESULTS: Garcinia buchananii bark extract inhibited pellet propulsion in a concentration-dependent manner, with an optimal concentration of ∼10 mg powder per mL Krebs. Interestingly, washout of the extract resulted in an increase in pellet propulsion to a level above basal activity. The extract reversibly reduced the amplitude of evoked fEPSPs in myenteric neurons. The extract's inhibitory action on propulsive motility and fEPSPs was not affected by the opioid receptor antagonist, naloxone, or the alpha- 2 adrenoceptor antagonist, yohimbine. The extract inhibited pellet motility in the presence of gamma-aminobutyric acid (GABA), GABA(A) and GABA(B) receptor antagonists picrotoxin and phaclofen, respectively. However, phaclofen and picrotoxin inhibited recovery rebound of motility during washout. CONCLUSIONS & INFERENCES: Garcinia buchananii extract has the potential to provide an effective, non-opiate antidiarrheal drug. Further studies are required to characterize bioactive components and elucidate the mechanisms of action, efficacy, and safety.