scholarly journals Grape-Seed Proanthocyanidins are Able to Reverse Intestinal Dysfunction and Metabolic Endotoxemia Induced by a Cafeteria Diet in Wistar Rats

Nutrients ◽  
2019 ◽  
Vol 11 (5) ◽  
pp. 979 ◽  
Author(s):  
Carlos González-Quilen ◽  
Katherine Gil-Cardoso ◽  
Iris Ginés ◽  
Raúl Beltrán-Debón ◽  
Montserrat Pinent ◽  
...  
Keyword(s):  
Wistar Rats ◽  
Ex Vivo ◽  
Ussing Chamber ◽  
Intestinal Barrier ◽  
Grape Seed ◽  
Cafeteria Diet ◽  
Tnf Α ◽  
Obesogenic Diet ◽  
Metabolic Endotoxemia

We evaluated the effectiveness of pharmacological doses of grape-seed proanthocyanidin extract (GSPE) in reversing intestinal barrier alterations and local inflammation in female Wistar rats fed a long-term obesogenic diet. Animals were fed a 17-week cafeteria diet (CAF diet), supplemented with daily GSPE doses (100 or 500 mg kg−1 body weight) during the final two weeks. CAF diet enhanced the intestinal permeation of an orally administered marker (ovalbumin, OVA) and increased the plasma levels of tumor necrosis factor-α (TNF-α) and lipopolysaccharides (LPS) in 2–3-fold. Ex vivo Ussing chamber assays showed a 55–70% reduction in transepithelial electrical resistance (TEER) and increased the TNF-α secretions in both small and large intestinal sections with a 25-fold increment in the ileum. Ileal tissues also presented a 4-fold increase of myeloperoxidase (MPO) activity. Both GSPE-treatments were able to restitute TEER values in the ileum and colon and to reduce plasma LPS to basal levels without a dose-dependent effect. However, effects on the OVA permeation and TNF-α secretion were dose and section-specific. GSPE also reduced ileal MPO activity and upregulated claudin 1 gene expression. This study provides evidence of the efficacy of GSPE-supplementation ameliorating diet-induced intestinal dysfunction and metabolic endotoxemia when administered at the end of a long-term obesogenic diet.

Inhaled Particulate Matter Affects Immune Responsiveness of Human Lung Phagocytes to Mycobacteria

2021 ◽  
Author(s):  
Brandon Anton Paarwater ◽  
Jomien Mouton ◽  
Samantha L Sampson ◽  
Stephanus T Malherbe ◽  
Jane A Shaw ◽  
...  
Keyword(s):  
Particulate Matter ◽  
Post Infection ◽  
Ex Vivo ◽  
Early Time ◽  
Immune Responsiveness ◽  
Phenotypic Profile ◽  
Host Protection ◽  
Tnf Α ◽  
Long Term Impact

The influence of smoke- or air pollution-derived cytoplasmic particulate matter (PM) can be detrimental and lead to failed lung immunity. We investigated mycobacterial uptake, intracellular replication, and soluble immune mediator responses of human bronchoalveolar lavage cells (BALC) loaded with/without PM, to infection with mycobacterial strains. We observed that only BALC containing PM display an ex vivo phenotypic profile dominated by spontaneous interleukin (IL) -10 production. PM loaded BALC retained the ability to phagocytose both Mycobacterium bovis Bacille Calmette Guérin (BCG) and Mycobacterium tuberculosis (M.tb) ΔleuDΔpanCD at equal efficacy as clear non-PM loaded BALC. However, immune responsiveness, such as the production of IL-6 (p=0.015) and tumor necrosis factor (TNF)-α (p= 0.0172) immediately post M.bovis BCG infection, were dramatically lower in black BALC loaded with PM versus clear non-PM loaded BALC. By 24 hour post infection, differential immune responses to M.bovis BCG between black versus clear BALC waned, and instead, production of IL-6 (p= 0.03) and IL-1α (p= 0.04 ) by black BALC were lower versus clear BALC following M.tb ΔleuDΔpanCD infection. Considering that TNFα and IL-6 are characterized as critical to host protection against mycobacteria, our findings suggest that BALC loaded with inhaled PM, display lower levels of anti-mycobacterial mediators, and that the response magnitude differs according to infective mycobacterial strain. Even though this did not translate into altered mycobacterial killing at early time points post infection, the long-term impact of such changes remains to be established.

scholarly journals Sex differences in morphofunctional changes in the immune system in Wistar rats of different age groups with experimental endotoxinemia

2020 ◽  
pp. 65-73
Author(s):  
Anna M. Kosyreva ◽  
Olga V. Makarova
Keyword(s):  
Gender Differences ◽  
Sex Differences ◽  
Immune System ◽  
Peripheral Blood ◽  
Wistar Rats ◽  
Ex Vivo ◽  
Age Groups ◽  
Mature Adult ◽  
Tnf Α ◽  
Ifn Γ

Objective. The aim was revealing gender differences in morphological and functional changes of lymphoid organs (thymus and spleen), changes of cytokine production and subpopulation composition of peripheral blood lymphocytes in Wistar rats of three age groups with endotoxemia. Materials and methods. We used male and female Wistar rats of three age groups: newborns, prepubertal and sexually mature adult rats. A day after the injection of 15 mg/kg of O26:B6 E. coli lipopolysaccharide (LPS), the volume fraction of the functional zones of the thymus and spleen, the number of AnnexinV + apoptotically dying cells in the thymus, the relative and absolute number of lymphocyte subpopulations (CD3+CD4+, CD3+CD8a+, CD4+CD25+Foxp3+, CD3-CD45R+) in peripheral blood and ex vivo production of IL-2, IL-4, IFN-γ and TNF-α were estimated. Results. Sex differences in the response of the immune system after the LPS injection in different age periods are expressed differently: in the neonatal period, there is immunosuppression in females (decrease in the ex vivo production of IL-2, TNF-α and IFN-γ), and there is activation of pro-inflammatory reactions in males (increase in ex vivo production of IL-2 and TNF-α). As compared with other age periods at prepubertal age, LPS-induced immunological disorders are more pronounced, and gender differences are minimal and related only to the number of T-regulatory and B-lymphocytes. In the adults, the LPS-induced immunosuppression is most pronounced in males - they have a decrease in the production of all the cytokines studied and a decrease in the number of cytotoxic and regulatory T-lymphocytes and B-cells. Conclusion. Thus, in each of the studied age periods - newborn, prepubertal and adult, the sexual differences in the immune system reactions are expressed differently and, apparently, these differences are determined by the content of sex steroid hormones, the concentration of which varies with age.

Generation of TNFα and Interleukin-6 by Peritoneal Macrophages after Overnight Dwells with Bicarbonate- or Lactate-Buffered Dialysis Fluid

2002 ◽  
Vol 22 (6) ◽  
pp. 663-669 ◽  
Author(s):  
Tomasz Liberek ◽  
Monika Lichodziejewska–Niemierko ◽  
Wanda Knopinska–Posluszny ◽  
Thomas P. Schaub ◽  
Judith Kirchgessner ◽  
...  
Keyword(s):  
Ex Vivo ◽  
Peritoneal Macrophages ◽  
Cytokine Release ◽  
Dialysis Fluid ◽  
Tnf Α ◽  
Peritoneal Dialysis Fluid ◽  
Initial Ph ◽  
Elisa Technique

Objective In order to evaluate the biocompatibility profile of a newly designed peritoneal dialysis fluid (PDF), we evaluated peritoneal leukocyte (PMΦ) cytokine release following overnight in vivo dwells using standard, lactate-buffered, single-chamber bag PDF (Lac-PDF) and purely bicarbonate-buffered, double-chamber bag PDF containing 34 (Bic-PDF) or 39 (Bic Hi-PDF) mmol/L bicarbonate. Design A randomized, open, crossover clinical trial with single weekly test dwells was performed in stable, long-term continuous ambulatory PD patients ( n = 8). During 8-hour overnight dwells, PMΦ were exposed to different PDF containing 1.5% glucose. After drainage, peritoneal cells were isolated and incubated with RPMI 1640 medium for 2 or 3 hours, with and without stimulation by lipopolysaccharide (LPS). Ex vivo release of tumor necrosis factor (TNF)-α and interleukin (IL)-6 was measured by specific ELISA technique. Results After pre-exposure to Lac-PDF, PMΦ generated 242 ± 279 pg TNFα/106 cells and 157 ± 105 pg IL-6/106 cells. When pre-exposed to Bic-PDF and Bic Hi-PDF, TNFα and IL-6 production of PMΦ was not significantly different from Lac-PDF. After LPS stimulation (100 ng/mL), PMΦ secretion of TNFα and IL-6 pre-exposed to three PDF revealed no significant differences between groups: TNFα was 2864 ± 1216, 2910 ± 1202, and 3291 ± 558 pg/106 cells after overnight dwells with Lac-PDF, Bic-PDF, and Bic Hi-PDF, respectively. Comparably, LPS-stimulated (100 pg/mL) PMΦ showed IL-6 secretion of 891 ± 335, 1380 ± 1149, and 1442 ± 966 pg/106 cells for Lac-PDF, Bic-PDF, and Bic Hi-PDF. Conclusion After long-term overnight dwells, initial pH, the different buffers, and varying glucose degradation product levels of PDF do not strongly affect PMΦ function with respect to cytokine release. The lack of significant differences between fluids may result from the complete dialysate equilibration achieved during the overnight intraperitoneal dwell.

scholarly journals PATHOPHYSIOLOGICAL CHANGES IN LIVER TISSUES OF HYPERCALORIE DIET-INDUCED OBESE RATS

2016 ◽  
Vol 10 (1) ◽  
pp. 338 ◽  
Author(s):  
Bharatha Ambadasu ◽  
Ramadevi S ◽  
Naikawadi Aa ◽  
Naveen Kumar M
Keyword(s):  
Metabolic Syndrome ◽  
Body Weight ◽  
Wistar Rats ◽  
Histopathological Examination ◽  
Cafeteria Diet ◽  
Nonalcoholic Fatty Liver ◽  
Obese Rats ◽  
Diet Intake ◽  
Liver Tissues

ABSTRACTObjectives: Hypercalorie diet intake has been associated with many long-term complications including metabolic syndrome, cardiovascular diseases,and nonalcoholic fatty liver disease.Methods: A total of 12 Wistar rats either sex were used in this study. These animals were randomly divided into two groups as control and obese rats.Group 1 consists of six rats weighing 150-200 g and fed with normal pellet chow. Another six rats were fed hypercalorie/cafeteria diet to induce obesity andincluded in the study after 19 weeks of age. All animals were sacrificed; liver tissues were collected, weighed and sent for the histopathological examination.Results: Weight of liver tissues of was significantly more in obese rats than the control rats. Histopathological examination shows an excessive fatdeposition and sinusoidal congestion in the liver tissues of obese rats.Conclusion: Increase in body weight is associated with the increase in fat deposition in the liver tissues which further develops into inflammationand necrosis of liver cells.Keywords: Wistar rats, Hypercalorie/cafeteria diet, Obese rats, Histopathological examination.

scholarly journals Continuous Dialysis with Bicarbonate/Lactate-Buffered Peritoneal Dialysis Fluids Results in a Long-Term Improvement in Ex Vivo Peritoneal Macrophage Function

2002 ◽  
Vol 13 (suppl 1) ◽  
pp. S97-S103
Author(s):  
Suzanne Jones ◽  
Clifford J. Holmes ◽  
Ruth K. Mackenzie ◽  
Rachel Stead ◽  
Gerald A. Coles ◽  
...  
Keyword(s):  
Peritoneal Dialysis ◽  
Peritoneal Macrophage ◽  
Ex Vivo ◽  
Negative Consequences ◽  
Necrosis Factor Alpha ◽  
Membrane Function ◽  
Neutral Ph ◽  
Tnf Α ◽  
Dialysis Fluids

ABSTRACT. To circumvent the potentially negative consequences of long-term exposure to unphysiologic acidic lactate-buffered peritoneal dialysis fluids (PDF), neutral pH solutions buffered with bicarbonate/lactate have recently been introduced in phase 2 and 3 clinical trials. This study examines the longitudinal changes in peritoneal macrophage (PMØ) function in patients dialyzed continuously with either lactate (LPD; 40 mM lactate, pH 5.2)-buffered or bicarbonate/lactate (TBL; 25 mM/15 mM bicarbonate/lactate, pH 7.3)-buffered PDF. Before the study, during the run in period of a phase 3 clinical trial, all patients had been taking LPD for at least the previous 18 wk. At the beginning of the study (day 0), both constitutive and serum-treated zymosan (STZ) stimulated tumor necrosis factor alpha (TNF-α) synthesis were assessed in PMØ isolated from 12-h dwell effluent (with 1.36% glucose) in all patients. The patients were subsequently randomized to either continuous TBL or LPD therapy and PMØ function was assessed after further 3- and 6-mo periods in all patients. At all time points measured STZ induced a dose-dependent increase in PMØ TNF-α secretion (P = 0.043 versus control for doses greater than 100 μg/ml). In patients continuously dialyzed with LPD, constitutive PMØ TNF-α synthesis levels (mean ± SEM, pg/106 PMØ per18 h, n = 5 patients) were 154 ± 65, 261 ± 60, and 101 ± 99 at 0, 3, and 6 mo, respectively. Stimulated STZ (1000 μg/ml) levels were 1340 ± 519, 1046 ± 586, and 758 ± 250 at 0, 3, and 6 mo, respectively. In patients dialyzed with TBL, constitutive PMØ TNF-α synthesis levels (pg/106 PMØ per 18 h, n = 5 patients) were 300 ± 136, 106 ± 35, and 213 ± 62 at 0, 3, and 6 mo, respectively. Stimulated STZ (1000 μg/ml) levels were 1969 ± 751, 1541 ± 330, and 2670 ± 671 at 0, 3, and 6 mo, respectively. At 6 mo, STZ-stimulated PMØ TNF-α synthesis was significantly higher in patients treated with TBL compared with those treated with LPD (P = 0.0035). These data suggest that in patients continuously dialyzed with a neutral pH solution, there is a long-term improvement in PMØ function compared with patients on conventional therapy. Better PMØ function suggests improved host defense status and may affect the peritoneum’s susceptibility to infection and potentially reduce the negative consequences of repeated intraperitoneal inflammation on long-term membrane function.

scholarly journals Acute Effects of Butyrate on Induced Hyperpermeability and Tight Junction Protein Expression in Human Colonic Tissues

Biomolecules ◽  
2020 ◽  
Vol 10 (5) ◽  
pp. 766
Author(s):  
Mathias W. Tabat ◽  
Tatiana M. Marques ◽  
Malin Markgren ◽  
Liza Löfvendahl ◽  
Robert J. Brummer ◽  
...  
Keyword(s):  
Tight Junction ◽  
Ex Vivo ◽  
Ussing Chamber ◽  
Intestinal Barrier ◽  
Intestinal Barrier Function ◽  
Healthy Humans ◽  
Junction Protein ◽  
Pre Treatment ◽  
Related Proteins

Intact intestinal barrier function is essential for maintaining intestinal homeostasis. A dysfunctional intestinal barrier can lead to local and systemic inflammation through translocation of luminal antigens and has been associated with a range of health disorders. Butyrate, a short-chain fatty acid derived from microbial fermentation of dietary fibers in the colon, has been described as an intestinal barrier-strengthening agent, although mainly by using in vitro and animal models. This study aimed to investigate butyrate’s ability to prevent intestinal hyperpermeability, induced by the mast cell degranulator Compound 48/80 (C48/80), in human colonic tissues. Colonic biopsies were collected from 16 healthy subjects and intestinal permeability was assessed by Ussing chamber experiments. Furthermore, the expression levels of tight junction-related proteins were determined by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Pre-treatment with 5 mM butyrate or 25 mM butyrate did not protect the colonic tissue against induced paracellular or transcellular hyperpermeability, measured by FITC-dextran and horseradish peroxidase passage, respectively. Biopsies treated with 25 mM butyrate prior to stimulation with C48/80 showed a reduced expression of claudin 1. In conclusion, this translational ex vivo study did not demonstrate an acute protective effect of butyrate against a chemical insult to the intestinal barrier in healthy humans.

scholarly journals Long-Lasting Effects of GSPE on Ileal GLP-1R Gene Expression Are Associated with a Hypomethylation of the GLP-1R Promoter in Female Wistar Rats

Biomolecules ◽  
2019 ◽  
Vol 9 (12) ◽  
pp. 865 ◽  
Author(s):  
Iris Ginés ◽  
Katherine Gil-Cardoso ◽  
Claudio D’Addario ◽  
Anastasia Falconi ◽  
Fabio Bellia ◽  
...  
Keyword(s):  
Gene Expression ◽  
Transcription Factor ◽  
Wistar Rats ◽  
Gene Promoter ◽  
Grape Seed ◽  
Cafeteria Diet ◽  
Sp1 Transcription Factor ◽  
Binding Factor ◽  
Female Wistar Rats ◽  
Grape Seed Proanthocyanidin

Flavonoids have been shown to modulate GLP-1 in obesity. GLP-1 induces some of its effects through the intestinal GLP-1 receptor (GLP-1R), though no data exist on how flavonoids affect this receptor. Here, we examine how a dose of grape seed proanthocyanidin extract (GSPE) with anti-obesity activity affects intestinal GLP-1R and analyze whether epigenetics play a role in the long-lasting effects of GSPE. We found that 10-day GSPE administration prior to the cafeteria diet upregulated GLP-1R mRNA in the ileum 17 weeks after the GSPE treatment. This was associated with a hypomethylation of the GLP-1R promoter near the region where the SP1 transcription factor binds. In the colon, the cafeteria diet upregulated GLP-1R without showing any GSPE effect. In conclusion, we have identified long-lasting GSPE effects on GLP-1R gene expression in the ileum that are partly mediated by hypomethylation at the gene promoter and may affect the SP1 binding factor.

Zinc strengthens the jejunal barrier by reversibly tightening the paracellular route

2017 ◽  
Vol 313 (6) ◽  
pp. G537-G548 ◽  
Author(s):  
Silke S. Zakrzewski ◽  
Michael Fromm ◽  
Jörg D. Schulzke ◽  
Dorothee Günzel
Keyword(s):  
Barrier Function ◽  
Ex Vivo ◽  
Intestinal Barrier ◽  
Secretory Diarrhea ◽  
Intestinal Barrier Function ◽  
Epithelial Barrier ◽  
Gastrointestinal Infections ◽  
Apical Side

During the postweaning period, piglets are prone to gastrointestinal infections. The resulting impairment of intestinal barrier function may cause diarrhea associated with growth retardation or even death of piglets. Orally applied Zn is commonly used to prevent and treat diarrhea, but its mode of action still needs to be elucidated. To analyze the molecular mechanism whereby Zn acts on porcine intestinal barrier function, ex vivo studies on piglet jejunum and accompanying in vitro studies on a porcine jejunal epithelial cell line, IPEC-J2/PS, were performed with electrophysiological tools. Feeding pharmacological Zn doses exerted no significant electrophysiologically ascertainable short- and long-term effects on jejunal barrier function ex vivo. However, in IPEC-J2/PS, basolateral Zn was cytotoxic since its application caused a release of lactate dehydrogenase and an irreversible breakdown of the epithelial barrier. In contrast, apical Zn application caused an immediate increase in paracellular resistance and a decrease in permeability to the paracellular marker fluorescein, reflecting overall barrier strengthening in vitro. Apical effects were fully reversible upon washout. This indicates that Zn supplemented to feed was completely washed out during ex vivo jejunum preparation. We conclude that there is no evidence for long-term barrier effects through prophylactic Zn supplementation and that extracellular Zn acts acutely and reversibly from the apical side via tightening the paracellular route, thus counteracting leak-flux diarrhea. NEW & NOTEWORTHY Therapeutically administered Zn successfully treats diarrhea in veterinary and human medicine. Here we present data that Zn strengthens the porcine jejunal epithelial barrier by reversibly tightening the paracellular route for inorganic ions and small solutes. Acute or long-lasting Zn effects on transcellular transport (Cl− secretion) were not detected. We therefore conclude that Zn is useful for acutely treating leak-flux diarrhea rather than secretory diarrhea. Suitability as prophylactic feed supplement, however, is questionable.

scholarly journals Long-term high-yield skeletal muscle stem cell expansion through staged perturbation of cytokine signaling in a soft hydrogel culture platform

2020 ◽  
Author(s):  
Alexander M. Loiben ◽  
Kun Ho Kim ◽  
Sharon Y. Soueid-Baumgarten ◽  
Victor M. Aguilar ◽  
Jonathan Chin Cheong ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Ex Vivo ◽  
High Yield ◽  
Muscle Stem Cell ◽  
Muscle Stem Cells ◽  
Stem Cell Expansion ◽  
Cytokine Cocktail ◽  
Tnf Α

AbstractMuscle stem cells (MuSCs) are an essential stem cell population for skeletal muscle homeostasis and regeneration throughout adulthood. MuSCs are an ideal candidate for cell therapies for chronic and acute muscle injuries and diseases given their inherent ability to self-renew and generate progenitor cells capable of myogenic commitment and fusion. Given their rarity and propensity to lose stem-cell potential in prolonged culture, methods for ex vivo MuSC expansion that achieve clinical-scale stem cell yields represent a critical unmet need in muscle cell-therapeutic development. Here, we tested a microenvironment engineering approach to achieve long-term adult mouse MuSC expansion suitable for clinical demands through the combined optimization of techniques previously reported to achieve small-yield MuSC expansion in short-term cultures. We developed an optimized protocol for high-yield MuSC expansion through the combination of inflammatory cytokine and growth factor co-stimulation, temporally-staged inhibition of the p38α/β mitogen activated protein kinase signaling pathway, and modulation of substrate rigidity in long-term hydrogel cultures. We found that, on soft, muscle-mimicking (12 kPa) hydrogel substrates, a mixture of the cytokines TNF-α, IL-1α, IL-13, and IFN-γ and the growth factor FGF2 stimulated robust exponential proliferation of adult MuSCs from both wildtype and mdx dystrophic mice for up to five weeks of culture that was accompanied by a phenotype shift towards committed myocytes. After observing that the temporal variation in myogenic commitment coincided with an oscillatory activation of p38α/β signaling, we tested a late-stage p38α/β inhibition strategy and found that blocking p38α/β signaling after three weeks, but not earlier, substantially enhanced cell yield, stem-cell phenotypes, and, critically, preserved transplantation potential for up to five weeks of FGF2/cytokine mix culture on soft hydrogels. Notably, this retention of transplant engraftment potency was not observed on traditional plastic substrates. We estimate that this protocol achieves >108-fold yield in Pax7+ stem cells from each starting MuSC, which represents a substantial improvement in stem-cell yield from long-term cultures compared to established methods.HighlightsTNF-α/IL-1α/IL-13/IFN-γ cytokine cocktail supports prolonged MuSC proliferation ex vivo but induces differentiation.Cytokine cocktail regulates cell signaling with varied prolonged activation signatures.Effects of p38α/β inhibition on cytokine-induced MuSC expansion are stage-dependent.Soft hydrogels with late-stage p38α/β inhibition expand functional Pax7+ MuSCs long-term.Short summaryCosgrove and colleagues develop a long-term muscle stem cell expansion protocol by combining a tunable stiffness hydrogel substrate, an inflammatory cytokine cocktail, and targeted inhibition of p38 MAPK signaling. They show that soft, muscle-mimicking hydrogels with delayed p38 inhibition yield robust quantities of Pax7+ functional muscle stem cells.

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