Ussing Chamber System Research Articles

Rifaximin alleviates MCD diet-induced NASH in mice by restoring the gut microbiota and intestinal barrier

AimsDue to the increasing global incidence rate of nonalcoholic steatohepatitis (NASH) combined with the lack of effective treatment methods for this disease, there is an urgent need to find new treatment strategies. The aim of this study was to investigate the efficacy of rifaximin in preventing and treating NASH and the related mechanism. Materials and methodsA NASH model was constructed by feeding male C57BL/6 mice a methionine-choline-deficient (MCD) diet for 4 weeks. Rifaximin was administered for 1 week before MCD diet feeding or during the last week of MCD diet feeding to investigate its preventive or therapeutic effects. Liver pathology, hepatic enzyme levels and metabolic indices were measured to evaluate the effects of rifaximin on NASH. Intestinal barrier integrity was measured via the Ussing chamber system and western blotting. 16S rDNA sequencing was conducted to investigate the fecal microbiota composition. Western blotting was performed to evaluate peroxisome proliferator activated receptor (PPAR)α and PPARγ protein levels. Key findingsRifaximin effectively alleviated MCD diet-induced NASH. The microbiota composition in MCD diet-fed mice was significantly altered, and intestinal barrier integrity was disrupted. Dysbiosis and intestinal barrier dysfunction were reversed by rifaximin. In addition, rifaximin modulated PPARα and PPARγ expression in the liver. SignificanceRifaximin effectively alleviated MCD diet-induced NASH by restoring the gut microbiota and reversing intestinal barrier dysfunction, suggesting that rifaximin treatment is a new approach for preventing and treating NASH.

Protective role of M3 muscarinic acetylcholine receptor in indomethacin-induced small intestinal injury.

EP4 prostanoid receptor (EP4R) contributes to the intestinal epithelial Cl- secretion, and inhibition of prostaglandin E (PGE) production by non-steroidal anti-inflammatory drugs (NSAIDs) plays a central role in NSAID-induced enteropathy. Although M3 muscarinic acetylcholine receptor (M3R) also contributes to the intestinal epithelial Cl- secretion, it remains unclear whether M3R is involved in NSAID-induced enteropathy due to a lack of selective agents. The present study explored how M3R is involved in the regulation of the intestinal epithelial Cl- secretion and its pathophysiological role in NSAID-induced enteropathy. Using the novel highly-selective M3 positive allosteric modulator PAM-369 that we recently developed, we evaluated the role of M3R in the intestinal epithelial secretion ex vivo by measuring the short circuit current (Isc) of intestinal epithelium with a Ussing chamber system and examined whether or not M3R protects against small intestinal injury in indomethacin-treated mice. Both the PGE1 derivative misoprostol and carbachol similarly increased the Isc in a concentration-dependent manner. The Isc increases were abolished either by receptor antagonists (an EP4R antagonist and a M3R antagonist, respectively) or by removal of extracellular Cl-. PAM-369 enhanced the carbachol-induced Isc by potentiating M3R, which could contribute to enhanced intestinal epithelial secretion. Treatment with PAM-369 ameliorated small intestinal injury in indomethacin-treated mice. Importantly, the M3R expression was significantly up-regulated, and PAM-369 potentiation of M3R was augmented in indomethacin-treated mice compared to untreated mice. These findings show that M3R plays a role in maintaining the intestinal epithelial secretion, which could contribute to protection against indomethacin-induced small intestinal injury. M3R is a promising target for treating or preventing NSAID-induced enteropathy. KEY MESSAGES: PAM-369, the M3 positive allosteric modulator, was used to potentiate M3R. PAM-369 enhanced carbachol-induced Isc in mouse ileum. PAM-369 ameliorated small intestinal injury in indomethacin-treated mice. M3R is a promising target for treating or preventing NSAID-induced enteropathy.

Role of CRF1 and CRF2 Receptors in Stress-induced Increase of Intestinal Epithelial Permeability in the Mouse Colon

Background and Aims: Stress is known to increase intestinal permeability, which may contribute to the development of irritable bowel syndrome (IBS) and other intestinal disorders. Corticotropin releasing factor (CRF) has been proven to mediate stress-induced changes in gut function, such as increased colonic motility, ion secretion, and intestinal permeability. Actions of CRF are mediated by two G-protein coupled receptors, CRF1 and CRF2; both are expressed in the intestine. However, the roles of the CRF1 and CRF2 receptors in stress-induced increase of intestinal epithelial permeability remain unknown. Methods: Adult male and female C57BL/6 mice were restrained (1h/d) for five consecutive days. The selective CRF1 and CRF2 receptor antagonists were injected intraperitoneally 15 min prior to the restraint stress. Control mice were kept in their home cages without restraint. After the last restraint stress/control session, the proximal colon was removed, and the mucosa/submucosa preparations were dissected and mounted onto the Ussing Chamber System for the measurement of intestinal permeability. Transepithelial resistance (TER) and FITC-inulin (4kD) flux across the colonic epithelium were used as indicators of paracellular permeability. Horseradish peroxidase (HRP, 40kD) flux was used to measure transcellular permeability. Results: Restraint stress did not alter TER or FITC-inulin flux in male mice. Pretreatment with the CRF1 receptor antagonist NBI 27914 or the CRF2 receptor antagonist antisauvagine-30 had no effect on TER or FITC-inulin flux in male mice. In female mice, restraint stress significantly reduced TER and increased FITC-inulin flux in the proximal colon, which were blocked by NBI 27914 but not by antisauvagine-30. Restraint stress significantly increased HRP flux in both male and female mice. In male mice, pretreatment with NBI 27914 significantly reduced stress-evoked increase of HRP flux, while antisauvagine-30 had no effect. In female mice, both NBI 27914 and antisauvagine-30 significantly reduced stress-evoked increase of HRP flux. Conclusions: Stress evokes different changes in epithelial permeability in the proximal colon of male and female mice. In male mice, stress does not influence paracellular permeability but increases transcellular permeability, which is mediated by the CRF1 receptor. In female mice, stress increases both transcellular and paracellular permeability. Stress-induced increase of paracellular permeability was mediated by the CRF1 receptor, but the increased transcellular permeability was mediated by both CRF1 and CRF2 receptors. University of Wisconsin-La Crosse Undergraduate Research Grant (CS and LB). This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

A sensorised, microfluidic Ussing chamber manufactured by stereolithography and soft lithography techniques

Motivated by the need for improved platforms in biomedical research, this study addresses challenges associated with traditional Ussing chamber systems, widely used in studying biological barriers like the epithelial barrier of the gut. These challenges include complexity, high sample volumes, and limited compatibility. By combining stereolithography and soft lithography techniques, a microfluidic Ussing chamber is manufactured, overcoming these limitations, and incorporating compatibility with microscopy. Validation through Trans-Epithelial Electrical Resistance (TEER) measurements confirms its efficacy in assessing ion permeability dynamics, utilizing Caco-2 cell monolayers. The study showcases the capability of the manufactured chamber for sensing impact of calcium on tight junctions.

An investigation of different antimycotoxin additives in swine intestinal explants challenged with aflatoxin and fumonisin: ex vivo and in vitro models

Abstract This study aimed to develop an ex vivo model to evaluate the efficacy of antimycotoxin additives (AMAs) against aflatoxin B1 (AFB1) and fumonisin B1 (FB1) using intestinal explants of growing pigs. Four assays of two treatments with 12 replicates each (24 jejunal explants used per assay) were performed using an Ussing Chamber (UC) system: two assays to evaluate two AMAs for AFB1 and two assays to evaluate two AMAs for FB1. The difference between the two assays for both AFB1 and FB1 was the composition of the additive used. The treatments for AFB1 assays were control [Buffer solution (BS) + 1 mg/l AFB1] and AMA (BS + 1 mg/l AFB1 + 0.5% AMA 1 or 2). The treatments for FB1 assays were control (BS + 50 mg/l FB1) and AMA (BS + 50 mg/l FB1 + 0.5% AMA 3 or 4). The efficacy of the four additives was also tested in vitro. The AFB1 concentrations in the explants from AMAs 1 and 2 were lower () than in the control. AMAs 1 and 2 reduced the jejunal absorption of AFB1 by 83.4 and 72.9%, respectively. Explants from AMAs 3 and 4 had lower FB1 () concentration when compared to the respective control treatment. AMAs 3 and 4 reduced the FB1 absorption by 31.9 and 17.6%, respectively. In the in vitro test, AMAs 1 and 2 provided 98.4 and 86.3% of AFB1 adsorption, respectively, while AMA 3 and 4 provided 91.2 and 80.5% FB1 adsorption, respectively. The ex vivo model can be a useful tool in evaluating the effectiveness of antimycotoxin additives for AFB1 and FB1 in swine. However, the low FB1 uptake in jejunal explants highlights the need for the development of additional information to improve the method.

Porphyromonas gingivalis Lipopolysaccharide Damages Mucosal Barrier to Promote Gastritis-Associated Carcinogenesis.

Recent epidemiological studies suggested correlation between gastric cancer (GC) and periodontal disease. We aim to clarify involvement of lipopolysaccharide of Porphyromonas gingivalis (Pg.), one of the red complex periodontal pathogens, in the GC development. To evaluate barrier function of background mucosa against the stimulations, we applied biopsy samples from 76 patients with GC using a Ussing chamber system (UCs). K19-Wnt1/C2mE transgenic (Gan) mice and human GC cell-lines ± THP1-derived macrophage was applied to investigate the role of Pg. lipopolysaccharide in inflammation-associated carcinogenesis. In the UCs, Pg. lipopolysaccharide reduced the impedance of metaplastic and inflamed mucosa with increases in mRNA expression of toll-like receptor (TLR) 2, tumor necrosis factor (TNF) α, and apoptotic markers. Invitro, Pg. lipopolysaccharide promoted reactive oxidative stress (ROS)-related apoptosis as well as activated TLR2-β-catenin-signaling on MKN7, and it increased the TNFα production on macrophages, respectively. TNFα alone activated TLR2-β-catenin-signaling in MKN7, while it further increased ROS and TNFα in macrophages. Under coculture with macrophages isolated after stimulation with Pg. lipopolysaccharide, β-catenin-signaling in MKN7 was activated with an increase in supernatant TNFα concentration, both of which were decreased by adding a TNFα neutralization antibody into the supernatant. In Gan mice with 15-week oral administration of Pg. lipopolysaccharide, tumor enlargement with β-catenin-signaling activation were observed with an increase in TNFα with macrophage infiltration. Local exposure of Pg. lipopolysaccharide may increase ROS on premalignant gastric mucosa to induce apoptosis-associated barrier dysfunction and to secrete TNFα from activated macrophages, and both stimulation of Pg. lipopolysaccharide and TNFα might activate TLR2-β-catenin-signaling in GC.

PSXI-23 Effects of Dietary Threonine, Tryptophan, and Glycine Supplementation on Growth Performance and Intestinal Barrier Function in Broiler Chickens Raised Under Multiple Stress Conditions

Abstract The objective of the current study was to compare the effects of dietary threonine, tryptophan, and glycine supplementation on growth performance and intestinal barrier function in broiler chickens raised under multiple stress conditions. All birds were allotted to 1 of 5 treatments with 8 replicates in a completely randomized design. Positive control (PC) treatment was normal condition and other treatments are multiple stress conditions with heat stress and high stocking density. Multiple stress conditions were exposed to a cyclic heat stress condition (30 ± 0.3°C for 10 h/d and 23 ± 0.2°C for 14 h/d). In addition, 10 birds per cage (16.9 birds/m2) were allotted in multiple stress conditions, but 15 birds per cage (25.3 birds/m2) were allotted in normal conditions. Basal diet was fed to broiler chickens in PC (normal condition) and negative control (NC; multiple stress conditions) treatments. Three additional diets were prepared by adding 0.68% threonine (99%), 0.19% tryptophan (99%), or 1.60% glycine (99%) to the basal diet. Four treatment diets such as NC, threonine, tryptophan, and glycine treatments were provided to birds in multiple stress conditions. All experimental diets and water were provided ad libitum for 14 d. The body weight gain (BWG) and feed intake (FI) were recorded at the end of the experiment. The feed efficiency (FE, g/kg) was calculated by dividing BWG with FI after mortality correction for FI. The intestinal barrier function was analyzed by trans-epithelial electrical resistance (TEER) value in a two-channel Ussing chamber system. Statistical analysis was conducted as a completely randomized design by using the PROC MIXED procedure of SAS (SAS Institute Inc., Cary, NC). The replicate was considered an experimental unit. Outlier data were confirmed by the UNIVARIATE procedure of SAS. The LSMEANS procedure was conducted to calculate treatment means. In addition, if the difference was significant, the PDIFF option of SAS was conducted to separate the means. The significance for statistical tests was considered at P < 0.05. Results indicated that all treatments under multiple stress condition (i.e., NC, Thr, Trp, and Gly treatments) caused a significant reduction (P < 0.05) in BW, BWG, FI, and FE as compared with PC treatment (Table 1). Dietary Thr supplementation under multiple stress conditions was similar FE compared with PC treatment (Table 2). Dietary threonine, tryptophan, and glycine supplementation under multiple stress condition induced intermediated values for TEER with dietary threonine supplementation having similar values to PC treatment. In conclusion, dietary supplementation of threonine seems the most effective functional amino acid. In addition, it can be a novel dietary treatment to improve growth performance and gut health in broiler chickens raised under multiple stress condition.

Novel co-crystal of 3-methylcinnamic acid with berberine (1:1): synthesis, characterization, and intestinal absorption property

Objective To synthesis a novel ‘Pharmaceutical Cocrystal’ of berberine (BBR) with coformer 3-methylcinnamic acid (3MCA) for increasing its solubility and intestinal absorption property. Significance BBR–HCl has poor liposolubility, difficulty in penetrating the cell membrane and absorption in the gastrointestinal tract, low bioavailability, and limited clinical application. A new cocrystal is formed by the interaction between 3-MCA and BBR through molecular interaction, which improves the physicochemical properties, intestinal absorption property, and hygroscopicity. Methods The solvent evaporation method was used to synthesize BCR–3MCA cocrystal. The physicochemical properties of the crystals were confirmed by different spectral techniques, i.e. by X-ray diffraction (PXRD, SXRD), thermogravimetry and differential thermal analysis (DSC, TGA), and scanning electron microscopy (SEM). Hygroscopicity of the cocrystal was evaluated by dynamic water vapor sorption (DVS). The intestinal absorption property was evaluated by the Ussing chamber system. Results BBR and 3MCA can be directly self-assembled into uniform co-crystal by hydrogen bonds and π–π stacking interactions. Compared with BBR–HCl, the solubility of BBR–3MCA cocrystal in polar solvents of water, methanol, ethanol, and isopropanol increased by 13.9, 1.5, 4.7, and 15.8 times, respectively. The apparent absorption and the absorption rate constants were increased by 7.7 and 5.6 times, respectively. Surprisingly, BBR–3MCA co-crystal almost had no hygroscopicity. Conclusion The absolute molecular structure of the co-crystal was further confirmed by single crystal X-ray diffraction. The hydrogen bonds drove the formation of X-like one-dimensional unit. Compared to the BBR–HCl, BBR–3MCA cocrystal displayed superior dissolution and solubility performance, improved physical–chemical properties and significantly improved intestinal absorption.

Allergic inflammation disrupts epithelial electrogenic electrolyte transport through cholinergic regulation in the mouse colon.

Intestinal transport of electrolytes is regulated by the enteric nervous system. Acetylcholine (ACh) is considered the most important neurotransmitter for electrolyte transport in the colon. However, electrolyte transport regulated by ACh is not fully understood in the colon. We investigated the regulation of electrogenic electrolyte transport by cholinergic agonists in the mouse colon by measuring the short-circuit current (Isc) using an Ussing chamber system. Muscarinic stimulation induced transient electrogenic Cl- secretion, and nicotinic stimulation induced electrogenic K+ secretion to the apical side in the normal mouse colon, and these effects were reduced in the colon of mice with food allergy (FA). Administration of prednisolone to mice with FA suppressed mild inflammation in the colon and allergic symptoms and thereby ameliorated the disruption of electrogenic electrolyte transport induced not only by cholinergic pathway activation but also by electrical field stimulation and intracellular cAMP signaling pathway activation in the colon. These results suggest that the electrogenic electrolyte transport function in the colon is impaired by FA-induced colonic inflammation and that the suppression of inflammation ameliorates the dysfunction of electrogenic electrolyte transport in the colon of mice with FA.

Similar in vitro response of rat brain nerve terminals, colon preparations and COLO 205 cells to smoke particulate matter from different types of wood

Major source of carbon-containing air born particular matter that significantly pollutes environment and provokes development of neuropathology is forest fires and wood combustion. Here, water-suspended smoke particulate matter preparations (SPs) were synthesized from birch, pine, poplar wood, and also birch bark and pine needles. Taking into account importance of the gut-brain communication system, SP properties were compared regarding their capability to modulate functioning of nerve terminals and gut cells/preparations. In cortex nerve terminals, poplar wood SP was more effective in decreasing uptake and increasing the extracellular levels of excitatory and inhibitory neurotransmitters L-[14C]glutamate and [3H]GABA, respectively. Spontaneous and H2O2-stimulated ROS generation in nerve terminals decreased by SPs, the most efficient one was from poplar wood. SPs from birch, pine and poplar wood caused membrane depolarization, poplar wood SP effect was 5-fold higher vs. birch and pine wood ones. Functional characteristics of gut cells/preparations, which tightly related to nerve terminal experiments, were assessed. SPs increased paracellular permeability of proximal colon mucosal-submucosal preparations monitored in Ussing chamber system (FITC-dextran, 4 kDa), where the most prominent effect had poplar wood SP. The latter demonstrated more considerable influence on COLO 205 cell causing 30 % loss of cell viability. PM emitted to the environment during combustion of various wood caused similar unidirectional harmful effects on brain and gut cell functioning, thereby triggering development of pathologies in gut and brain and gut-brain communication system.

Size-dependent transfer of microplastics across the intestinal wall of the echinoid Paracentrotus lividus.

The fate and toxicity of ingested marine microplastics (MPs) have been of major concern in aquatic ecotoxicology for the last decade. Although their ingestion by a wide range of marine organisms has been proven, the uptake of MPs within organs is not yet fully understood and relies on the ability of ingested microplastics to transfer from the gut to tissues beyond the digestive wall (i.e., translocation). The present study investigates the in vitro transfer of fluorescent high-density polyethylene particles of different sizes classes (1-5µm; 10-29µm; 38-45µm) across the intestinal wall of the sea urchin Paracentrotus lividus using Ussing chambers. Small microplastics (1-5µm) were proven to be able to cross the intestinal wall of P. lividus and reach the coelomic fluid, while larger microplastics (≥ 10µm) were not observed to cross the intestinal wall. Results demonstrate a size-dependent passage of polyethylene microparticles across the intestinal walls of P. lividus for the first time, highlighting the suitability of Ussing chamber systems to study the transfer of MPs across the intestinal wall of animals.

Comparision of the phenol red, gravimetric, and synthesized mPEG-PR methods for correcting water flux using the single-pass intestinal perfusion method

Phenol red and PEG-4000, the usual non-absorbable indicators, have non-negligible absorption problems in measuring water flux. mPEG-PR, combined phenol red with mPEG-4000, was first synthesized and could decrease absorption. However, its application has not been confirmed. The purpose of this study was to explore the applicability of mPEG-PR as a novel non-absorption indicator in the in situ single-pass intestinal perfusion (SPIP) experiment. Six model drugs (atenolol ranitidine, ibuprofen, ketoprofen, antipyrine, hydrochlorothiazide) were used to compare the accuracy of four measuring methods including phenol red, mPEG-PR, gravimetric, and non-corrected methods of correcting intestinal fluid transport. Moreover, we evaluated the correlations between the effective permeability coefficients (Peff) in rat and fraction dose absorbed (Fabs) in human, Peff in human, and apparent permeability coefficients (Papp) by the Ussing Chamber system using human tissue. Among these methods, mPEG-PR was the most reliable approach, which avoided the absorption of phenol red method and mucous shedding or water evaporation of gravimetric method. An excellent correlation was obtained between the Peff of rat and Fabs of human. Our results of this study indicated that mPEG-PR was a stable and accurate non-absorbable indicator to correct water flux in the in situ SPIP model, which could be developed to predict the human Fabs.

In vitro and in vivo evaluation of cinnamaldehyde Microemulsion-Mucus interaction.

The current investigation explores the possible mechanism of the microemulsion drug delivery system to improve the oral bioavailability of cinnamaldehyde (CA), an important food spice, from the perspective of the microemulsion-mucus system. The cinnamaldehyde microemulsion (CA-ME) was prepared by the water titration method combined with the pseudo-ternary phase diagram. The dynamic analysis was applied to detect the drug release in vitro. An intestinal mucosal injury test was conducted to evaluate the safety of CA-ME and drug absorption across the intestinal tract of rats was investigated through an Ussing chamber system. The rheology of blank mucus and drug-loaded mucus was investigated using a rheometer. The bioavailability of CA-ME in rats was evaluated through pharmacokinetic characteristics. The ratio of optimal prescription was Tween 80: 1,2-propanediol: vitamin E oil: CA: water=24.3:4.8:5:7.5:58.4. The droplets were uniform in size and evenly dispersed. Rheological studies showed that the microemulsion-mucus system all exhibit pseudoplastic fluid behavior, and CA-ME increased the viscosity of the mucus to a certain extent. Compared with CA solution, CA-ME promoted the absorption of CA in various intestinal segments, especially the ileum. Pharmacokinetic experiments showed that the relative bioavailability of CA-ME was enhanced 2.5-fold higher than that of CA solution. ME as a carrier for lipophobic substances, may increase the viscosity of the intestine mucus system to obtain longer residue time and better absorption. PRACTICAL APPLICATIONS: In this study, in vitro absorption Ussing model was combined with rheological and pharmacokinetic analysis to systematically analyze the intestinal mucus mechanism of microemulsion to improve the oral bioavailability of cinnamic aldehyde. It laid the foundation for exploring the absorption and transport of drugs in the intestinal mucus barrier.

Evaluation of an Ussing Chamber System Equipped with Rat Intestinal Tissues to Predict Intestinal Absorption and Metabolism in Humans.

Oral bioavailability (F) is one of the key factors that need to be determined in drug discovery. This factor is determined by the permeability and solubility of new molecule entities (NMEs) according to the biopharmaceutics classification system (BCS). In the present study, we evaluated the permeability of 22 drugs in rat intestinal tissues using an Ussing chamber system and correlated the permeability with data on human intestinal absorption (Fa) and intestinal availability (Fa×Fg) reported in the literature. The rat intestinal permeability data were better correlated with the combined effect of the absorbed fraction (Fa) and the fraction escaping intestinal metabolism (Fg) than Fa itself. Clear regional dependent absorption was observed for most of the test drugs, and ileal Papp was generally higher than that in other segments. Finally, the function of the efflux transporter P-glycoprotein (P-gp) with regard to oral absorption of substrates was evaluated with an Ussing chamber. We also demonstrated that the rat intestinal stability of the three cytochrome P450 (CYP) substrates was consistent with the human data. An Ussing chamber system incorporating rat intestinal tissue would be a valuable tool to predict human intestinal absorption and metabolism for molecules with various physicochemical properties.

Combination of mouse prion protein with detoxified lipopolysaccharide triggers colon genes related to inflammatory, antibacterial, and apoptotic responses

Previously we observed that bacterial lipopolysaccharide (LPS) was able to instantly convert recombinant murine prion protein (moPrP) from an alpha-helical to a beta-sheet enriched state. The objectives of this study were to evaluate the effects of a single in vitro administration of recombinant moPrP alone or combined with detoxified lipopolysaccharide (D-LPS) on innate immunity and antibacterial gene expression in the colon of male FVB/N mice, under an Ussing chamber system. Results showed that moPrP alone affected the expression of genes related to both toll-like receptor (TLR)- and nod-like receptor (NLR)-signaling as well as pro- and anti-inflammatory responses. moPrP induced a strong antibacterial response with Slpi mRNA over expression (> 9-fold). Combination of moPrP with D-LPS on the mucosal side of the colon induced genes associated with TLR-signaling, apoptosis, and a very strong antibacterial response (> 35-fold Slpi expression). Administration of moPrP on the mucosal side and D-LPS on the serosal side triggered expression of 12 genes related to TLR signaling, apoptosis, and antibacterial responses, as illustrated by overexpression of Slpi by >30-fold. The over expression of Slpi mRNA was further reaffirmed by ELISA and when moPrP was added to the mucosal side and D-LPS on the serosal side, an increased Slpi protein was observed. Application of combined moPrP and D-LPS on the mucosal side significantly increased the Slpi protein. Results of this study demonstrated that moPrP alone or combined with D-LPS affected the expression of various genes related to inflammation, antibacterial, and apoptotic responses.

Minimum biological domain of xenin-25 required to induce anion secretion in the rat ileum

Xenin-25 has a variety of physiological functions in the gastrointestinal tract, including ion transport and motility. Xenin-25 and neurotensin show sequence homology, especially near their C-terminal regions. The sequence similarity between xenin-25 and neurotensin indicates that the effects of xenin-25 is mediated by the neurotensin receptor but some biological actions of xenin-25 are independent. We have previously reported that xenin-25 modulates intestinal ion transport and colonic smooth muscle activity. However, minimal biological domain of xenin-25 to induce ion transport was not clear. To improve the mechanistic understanding of xenin-25 and to gain additional insights into the functions of xenin-25, the present study was designed to determine the minimal biological domain of xenin-25 required for ion transport in the rat ileum using various truncated xenin fragments and analogues in an Ussing chamber system. The present results demonstrate that the minimum biological domain of xenin-25 to induce Cl−/HCO3− secretion in the ileum contains the C-terminal pentapeptide. Furthermore, Arg at position 21 is important to retain the biological activity of xenin-25 and induces Cl−/HCO3− secretion in the rat ileum.

The Ussing Chamber System: Functioning and Determination of Intestinal Transepithelial Electrical Measurements and Intestinal Integrity in Broiler Chickens

Background: The epithelial barrier integrity reflects gut health in broiler chickens and impaired intestinal barrier is associated with production losses. So, the present report was undertaken to determine the transepithelial electrical measurements and intestinal integrity of intestinal tissues in broilers from retail shops in Bangalore, India using an Ussing chamber system. Methods: For the present study, commercial broiler chickens were purchased from registered retail meat shops, sacrificed and the intestinal tissues were collected. The transepithelial electrical voltage, short circuit current and transepithelial electrical resistance were measured in jejunum using a contemporary Ussing chamber system following methodologies as detailed in the manuscript. Result: The studied intestinal tissues exhibited transepithelial electrical movements across intestine and the integrity of intestinal tissues were maintained. The study also indicated the preservation of intestinal barrier integrity of broilers at the crucial destination of the live broiler chicken market which in turn ascertains the quality of the product offered to the customers.

Improved Hygroscopicity and Bioavailability of Solid Dispersion of Red Ginseng Extract with Silicon Dioxide.

This study aims to develop a powder formulation for the Korean red ginseng extract (RGE) and to evaluate its in vitro and in vivo formulation characteristics. The solid dispersion of RGE was prepared with hydrophilic carriers using a freeze-drying method. After conducting the water sorption–desorption isothermogram (relative humidity between 30 and 70% RH), differential scanning calorimetry thermal behavior, dissolution test, and intestinal permeation study, a solid dispersion formulation of RGE and silicon dioxide (RGE-SiO2) was selected. RGE-SiO2 formulation increased intestinal permeability of ginsenoside Rb1 (GRb1), GRb2, GRc, and GRd by 1.6-fold in rat jejunal segments as measured by the Ussing chamber system. A 1.6- to 1.8-fold increase in plasma exposure of GRb1, GRb2, GRc, and GRd in rats was observed following oral administration of RGE-SiO2 (375 mg/kg as RGE). No significant difference was observed in the time to reach maximum concentration (Tmax) and half-life in comparison to those in RGE administered rats (375 mg/kg). In conclusion, formulating solid dispersion of RGE with amorphous SiO2, the powder formulation of RGE was successfully formulated with improved hygroscopicity, increased intestinal permeability, and enhanced oral bioavailability and is therefore suitable for processing solid formulations of RGE product.

The impact of the intestinal microbiota and the mucosal permeability on three different antibiotic drugs

BackgroundThe totality of bacteria, protozoa, viruses and fungi that lives in the human body is called microbiota. Human microbiota specifically colonizes the skin, the respiratory and urinary tract, the urogenital tract and the gastrointestinal system. This study focuses on the intestinal microbiota to explore the drug-microbiota relationship and, therefore, how the drug bioavailability changes in relation to the microbiota biodiversity to identify more personalized therapies, with the minimum risk of side effects. MethodsTo achieve this goal, we developed a new mathematical model with two compartments, the intestine and the blood, which takes into account the colonic mucosal permeability variation - measured by Ussing chamber system on human colonic mucosal biopsies - and the fecal microbiota composition, determined through microbiota 16S rRNA sequencing analysis. Both of the clinical parameters were evaluated in a group of Irritable Bowel Syndrome patients compared to a group of healthy controls. Key ResultsThe results show that plasma drug concentration increases as bacterial concentration decreases, while it decreases as intestinal length decreases too. ConclusionsThe study provides interesting data since in literature there are not yet mathematical models with these features, in which the importance of intestinal microbiota, the ”forgotten organ”, is considered both for the subject health state and in the nutrients and drugs metabolism.

Impaired Colonic Contractility and Intestinal Permeability in Symptomatic Uncomplicated Diverticular Disease

Background/AimsImpaired intestinal motility seems to play a crucial role in symptomatic uncomplicated diverticular disease (SUDD), although the mechanism is not clear. The aim of the present study is to explore the contractility patterns of colonic smooth muscle strips (MS) and smooth muscle cells (SMCs) and to assess mucosal integrity in SUDD patients.MethodsMS or SMCs were isolated from specimens of human distal colon of 18 patients undergoing surgery for non-obstructive colonic cancer, among them 9 with SUDD. Spontaneous phasic contractions on strips and morpho-functional parameters on cells were evaluated in basal conditions and in response to acetylcholine (ACh). Mucosal integrity of SUDD colonic biopsies was evaluated by the Ussing Chamber system. Immunohistochemical staining for tight junction protein complex and for Toll-like receptor 4 (TLR4) was performed.ResultsColonic MS of SUDD group showed a significant reduced basal tone and ACh-elicited contraction, compared to the control group (9.5 g and 47.0% in the SUDD group; 14.16 g and 69.0% in the control group; P < 0.05). SMCs of SUDD group showed a maximal contractile response to ACh significantly reduced compared to control group (8.8% vs 16.5%, P < 0.05). SUDD patients displayed lower transepithelial electrical resistance and increased paracellular permeability compared to control group. Immunohistochemical expression of TLR4 was not different in both groups, while tight junction protein complex expression was lower in SUDD patients compared to control group patients.ConclusionIt could be hypothesized that in SUDD, in absence of severe inflammation, an increased intestinal mucosal permeability is related to altered colonic motility probably responsible for symptoms genesis.