IFN-γ and TNF-α decrease serotonin transporter function and expression in Caco2 cells

Recent studies have shown that mucosal serotonin (5-HT) transporter (SERT) expression is decreased in animal models of colitis, as well as in the colonic mucosa of humans with ulcerative colitis and irritable bowel syndrome. Altered SERT function or expression may underlie the altered motility, secretion, and sensation seen in these inflammatory gut disorders. In an effort to elucidate possible mediators of SERT downregulation, we treated cultured colonic epithelial cells (Caco2) with conditioned medium from activated human lymphocytes. Application of the conditioned medium caused a decrease in fluoxetine-sensitive [3H]5-HT uptake. Individual proinflammatory agents were then tested for their ability to affect uptake. Cells were treated for 48 or 72 h with PGE2 (10 μM), IFN-γ (500 ng/ml), TNF-α (50 ng/ml), IL-12 (50 ng/ml), or the nitric oxide-releasing agent S-nitrosoglutathione (GSNO; 100 μM). [3H]5-HT uptake was then measured. Neither PGE nor IL-12 had any effect on [3H]5-HT uptake, and GSNO increased uptake. However, after 3-day incubation, both TNF-α and IFN-γ elicited significant decreases in SERT function. Neither TNF-α nor IFN-γ were cytotoxic when used for this period of time and at these concentrations. These two cytokines also induced decreases in SERT mRNA and protein levels. By altering SERT expression, TNF-α and IFN-γ could contribute to the altered motility and expression seen in vivo in ulcerative colitis or irritable bowel syndrome.

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In Vivo Regulation of Replicative Legionella pneumophila Lung Infection by Endogenous Interleukin-12

Infection and Immunity ◽

10.1128/iai.66.1.65-69.1998 ◽

1998 ◽

Vol 66 (1) ◽

pp. 65-69 ◽

Cited By ~ 52

Author(s):

J. K. Brieland ◽

D. G. Remick ◽

M. L. LeGendre ◽

N. C. Engleberg ◽

J. C. Fantone

Keyword(s):

Legionella Pneumophila ◽

Lung Infection ◽

Interleukin 12 ◽

Necrosis Factor Alpha ◽

Protein Levels ◽

Tnf Α ◽

Factor Alpha ◽

Ifn Γ

ABSTRACT The in vivo role of endogenous interleukin 12 (IL-12) in modulating intrapulmonary growth of Legionella pneumophila was assessed by using a murine model of replicative L. pneumophila lung infection. Intratracheal inoculation of A/J mice with virulent bacteria (106 L. pneumophilacells per mouse) resulted in induction of IL-12, which preceded clearance of the bacteria from the lung. Inhibition of endogenous IL-12 activity, via administration of IL-12 neutralizing antiserum, resulted in enhanced intrapulmonary growth of the bacteria within 5 days postinfection (compared to untreated L. pneumophila-infected mice). Because IL-12 has previously been shown to modulate the expression of cytokines, including gamma interferon (IFN-γ), tumor necrosis factor alpha (TNF-α), and IL-10, which regulate L. pneumophila growth, immunomodulatory effects of endogenous IL-12 on intrapulmonary levels of these cytokines during replicative L. pneumophila lung infection were subsequently assessed. Results of these experiments demonstrated that TNF-α activity was significantly lower, while protein levels of IFN-γ and IL-10 in the lung were similar, in L. pneumophila-infected mice administered IL-12 antiserum, compared to similarly infected untreated mice. Together, these results demonstrate that IL-12 is critical for resolution of replicativeL. pneumophila lung infection and suggest that regulation of intrapulmonary growth of L. pneumophila by endogenous IL-12 is mediated, at least in part, by TNF-α.

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Bifico Relieves Irritable Bowel Syndrome By Regulating Gut Microbiota Dysbiosis and Inflammatory Cytokines in an Animal Model

10.21203/rs.3.rs-717837/v1 ◽

2021 ◽

Author(s):

Yanlin Zhou ◽

Fan Zhang ◽

Liqi Mao ◽

Tongfei Feng ◽

Kaijie Wang ◽

...

Keyword(s):

Irritable Bowel Syndrome ◽

Gut Microbiota ◽

Inflammatory Cytokines ◽

Short Chain Fatty Acids ◽

Intragastric Administration ◽

Protein Levels ◽

Tnf Α ◽

Irritable Bowel ◽

Factor Α ◽

Gut Microbiota Dysbiosis

Abstract Gut microbiota dysbiosis, a core pathophysiology of irritable bowel syndrome (IBS), is closely related to immunological and metabolic functions. Gut microbiota-based therapeutics have been recently explored in several studies. Bifico is a probiotic cocktail widely used in gastrointestinal disorders which relate to the imbalance of gut microbiota. However, the efficacy and potential mechanisms of Bifico treatment in IBS remains incompletely understood. In this animal experiment, IBS mice received Bifico by intragastric administration. Subsequently, abdominal withdrawal reflex (AWR) scores showed a protective effect of Bifico in IBS mice. Then 16S rDNA, 1H nuclear magnetic resonance (1H-NMR) and western blot assays were performed to analyze alterations of gut microbiota, microbiome metabolites and inflammatory cytokines, respectively. Results suggested that while Bifico did not increase gut microbial diversity, it could change the composition of gut microbiota which were characterized by an increase of Proteobacteria phylum and Actinobacteria phylum, Muribaculum genera, Bifidobacterium genera and a decrease of Parabacteroides genera, Sutterella genera and Lactobacillus genera. Moreover, Bifico elevated the concentration of short-chain fatty acids (SCFAs) and reduced protein levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α). From further Spearman's correlation analysis, Bifidobacterium genera were positively correlated with SCFAs including propionate, butyrate, valerate and negatively correlated with IL-6 and TNF-α. In conclusion, this study demonstrated that Bifico could alleviate symptoms of IBS mice through regulation of the gut microbiota, elevating production of SCFAs and reducing the colonic inflammatory response. Therefore, Bifico may have utility in clinical practice.

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Increased Colonic Epithelial Permeability and Mucosal Eosinophilia in Ulcerative Colitis in Remission Compared With Irritable Bowel Syndrome and Health

Inflammatory Bowel Diseases ◽

10.1093/ibd/izz328 ◽

2020 ◽

Vol 26 (7) ◽

pp. 974-984 ◽

Cited By ~ 1

Author(s):

Georgios Katinios ◽

Maite Casado-Bedmar ◽

Susanna A Walter ◽

Maria Vicario ◽

Ana M González-Castro ◽

...

Keyword(s):

Ulcerative Colitis ◽

Irritable Bowel Syndrome ◽

Mast Cells ◽

Ethylenediaminetetraacetic Acid ◽

Ussing Chamber ◽

Barrier Dysfunction ◽

Ussing Chambers ◽

Tnf Α ◽

51Cr Edta ◽

Irritable Bowel

Abstract Background Barrier dysfunction is recognized as a pathogenic factor in ulcerative colitis (UC) and irritable bowel syndrome (IBS), but it is unclear to what extent the factors related to barrier dysfunction are disease-specific. The aim of this study was to compare these aspects in UC patients in remission, IBS patients, and healthy controls (HCs). Methods Colonic biopsies were collected from 13 patients with UC in remission, 15 patients with IBS-mixed, and 15 HCs. Ulcerative colitis patients had recently been treated for relapse, and biopsies were taken from earlier inflamed areas. Biopsies were mounted in Ussing chambers for measurements of intestinal paracellular permeability to 51chromium (Cr)-ethylenediaminetetraacetic acid (EDTA). In addition, biopsies were analyzed for mast cells and eosinophils by histological procedures, and plasma tumor necrosis factor (TNF)-α was assessed by ELISA. Results Ussing chamber experiments revealed an increased 51Cr-EDTA permeability in UC and IBS (P < 0.05). The 51Cr-EDTA permeability was higher in UC compared with IBS (P < 0.005). There were increased numbers of mucosal mast cells and eosinophils in UC and IBS and more eosinophils in UC compared with IBS (P < 0.05). Also, increased extracellular granule content was found in UC compared with HCs (P < 0.05). The 51Cr-EDTA permeability correlated significantly with eosinophils in all groups. Plasma TNF-α concentration was higher in UC compared with IBS and HCs (P < 0.0005). Conclusions Results indicate a more permeable intestinal epithelium in inactive UC and IBS compared with HCs. Ulcerative colitis patients, even during remission, demonstrate a leakier barrier compared with IBS. Both eosinophil numbers and activation state might be involved in the increased barrier function seen in UC patients in remission.

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A Clostridia-rich microbiota contributes to increased excretion of bile acids in diarrhea-predominant irritable bowel syndrome

10.1101/630350 ◽

2019 ◽

Author(s):

Ling Zhao ◽

Wei Yang ◽

Yang Chen ◽

Fengjie Huang ◽

Lin Lu ◽

...

Keyword(s):

Irritable Bowel Syndrome ◽

Bile Acids ◽

Protein Levels ◽

Human Microbiota ◽

Fecal Bile Acids ◽

Fgf19 Expression ◽

Irritable Bowel ◽

Fibroblast Growth Factor 19

ABSTRACTObjectiveAn excess of fecal bile acids (BAs) is thought to be one of the mechanisms for diarrhea-predominant irritable bowel syndrome (IBS-D). However, the factors causing excessive BA excretion remains unclear. Given the importance of gut microbiota in BA metabolism, we hypothesized that gut dysbiosis might contribute to excessive BA excretion in IBS-D.DesignMetabolomic and metagenomic analyses were performed of specimens from 290 IBS-D patients and 89 healthy volunteers. By transplanting human microbiota and manipulating specific microbiome species in mice, the effects of microbiota on host BA metabolism were assessed at metabolic, genetic and protein levels. Effects of individual and mixed BAs on enterohepatic feedback pathways were also tested in vitro and in vivo.ResultsTotal fecal BAs were excessively excreted in 24.5% of IBS-D patients. Their fecal metagenomes showed increased abundances of Clostridia and BA-transforming genes (hdhA and bais). The increases of Clostridia bacteria (e.g. C. scindens) were positively associated with the levels of fecal BAs and serum 7α-hydroxy-4-cholesten-3-one (C4), while being negatively correlated with serum fibroblast growth factor 19 (FGF19). Both Clostridia-rich human microbiota and C. scindens enhanced levels of serum C4 and hepatic conjugated BAs in mice recipients and reduced ileal FGF19 expression. Inhibition of Clostridium species by vancomycin yielded opposite findings. Clostridia-derived BAs (e.g. conjugated and free ursodeoxycholic acid) significantly suppressed intestinal FGF19 expression.ConclusionThe Clostridia-rich microbiota contributes to excessive BA excretion in IBS-D patients. This study provided the basis for more precise clinical diagnosis and management for IBS-D.

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Gut permeability in patients with irritable bowel syndrome and inactive ulcerative colitis

Zeitschrift für Gastroenterologie ◽

10.1055/s-0029-1224004 ◽

2009 ◽

Vol 47 (05) ◽

Author(s):

K Gecse ◽

R Róka ◽

T Séra ◽

A Annaházi ◽

A Rosztóczy ◽

...

Keyword(s):

Ulcerative Colitis ◽

Irritable Bowel Syndrome ◽

Gut Permeability ◽

Irritable Bowel

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Skullcapflavone II Suppresses TNF-α/IFN-γ-Induced TARC, MDC, and CTSS Production in HaCaT Cells

International Journal of Molecular Sciences ◽

10.3390/ijms22126428 ◽

2021 ◽

Vol 22 (12) ◽

pp. 6428

Author(s):

Hanon Lee ◽

Dong Hun Lee ◽

Jang-Hee Oh ◽

Jin Ho Chung

Keyword(s):

P38 Mapk ◽

Therapeutic Potential ◽

Inflammatory Diseases ◽

Mapk Signaling ◽

Hacat Cells ◽

Protein Levels ◽

Tnf Α ◽

Ifn Γ ◽

Mapk Signaling Pathways ◽

Pro Inflammatory Cytokines

Skullcapflavone II (SFII), a flavonoid derived from Scutellaria baicalensis, has been reported to have anti-inflammatory properties. However, its therapeutic potential for skin inflammatory diseases and its mechanism are unknown. Therefore, this study aimed to investigate the effect of SFII on TNF-α/IFN-γ-induced atopic dermatitis (AD)-associated cytokines, such as thymus- and activation-regulated chemokine (TARC) and macrophage-derived chemokine (MDC). Co-stimulation with TNF-α/IFN-γ in HaCaT cells is a well-established model for induction of pro-inflammatory cytokines. We treated cells with SFII prior to TNF-α/IFN-γ-stimulation and confirmed that it significantly inhibited TARC and MDC expression at the mRNA and protein levels. Additionally, SFII also inhibited the expression of cathepsin S (CTSS), which is associated with itching in patients with AD. Using specific inhibitors, we demonstrated that STAT1, NF-κB, and p38 MAPK mediate TNF-α/IFN-γ-induced TARC and MDC, as well as CTSS expression. Finally, we confirmed that SFII significantly suppressed TNF-α/IFN-γ-induced phosphorylation of STAT1, NF-κB, and p38 MAPK. Taken together, our study indicates that SFII inhibits TNF-α/IFN-γ-induced TARC, MDC, and CTSS expression by regulating STAT1, NF-κB, and p38 MAPK signaling pathways.

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Inflammatory cytokines-stimulated human muscle stem cells ameliorate ulcerative colitis via the IDO-TSG6 axis

Stem Cell Research & Therapy ◽

10.1186/s13287-020-02118-3 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Shengchao Zhang ◽

Jiankai Fang ◽

Zhanhong Liu ◽

Pengbo Hou ◽

Lijuan Cao ◽

...

Keyword(s):

Ulcerative Colitis ◽

Stem Cells ◽

Mesenchymal Stem Cells ◽

Inflammatory Cytokines ◽

Human Muscle ◽

Enzyme Linked Immunosorbent Assay ◽

Muscle Stem Cells ◽

Tnf Α ◽

Ifn Γ ◽

Anti Inflammatory

Abstract Background Muscle stem cells (MuSCs) are absolutely required for the formation, repair, and regeneration of skeletal muscle tissue. Increasing evidence demonstrated that tissue stem cells, especially mesenchymal stem cells (MSCs), can exert therapeutic effects on various degenerative and inflammatory disorders based on their immunoregulatory properties. Human mesenchymal stem cells (hMSCs) treated with interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) were reported to possess anti-inflammatory functions by producing TNF-stimulated gene 6 (TSG-6). However, whether human muscle stem cells (hMuSCs) also possess TSG-6 mediated anti-inflammatory functions has not been explored. Methods The ulcerative colitis mouse model was established by subjecting mice to dextran sulfate sodium (DSS) in drinking water for 7 days. hMuSCs were pretreated with IFN-γ and TNF-α for 48 h and were then transplanted intravenously at day 2 of DSS administration. Body weights were monitored daily. Indoleamine 2,3-dioxygenase (IDO) and TSG-6 in hMuSCs were knocked down with short hairpin RNA (shRNA) and small interfering RNA (siRNA), respectively. Colon tissues were collected for length measurement and histopathological examination. The serum level of IL-6 in mice was measured by enzyme-linked immunosorbent assay (ELISA). Real-time PCR and Western blot analysis were performed to evaluate gene expression. Results hMuSCs treated with inflammatory factors significantly ameliorated inflammatory bowel disease (IBD) symptoms. IDO and TSG-6 were greatly upregulated and required for the beneficial effects of hMuSCs on IBD. Mechanistically, the tryptophan metabolites, kynurenine (KYN) or kynurenic acid (KYNA) produced by IDO, augmented the expression of TSG-6 through activating their common receptor aryl hydrocarbon receptor (AHR). Conclusion Inflammatory cytokines-treated hMuSCs can alleviate DSS-induced colitis through IDO-mediated TSG-6 production.

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Tailored Hydrogels as Delivery Platforms for Conditioned Medium from Mesenchymal Stem Cells in a Model of Acute Colitis in Mice

Pharmaceutics ◽

10.3390/pharmaceutics13081127 ◽

2021 ◽

Vol 13 (8) ◽

pp. 1127

Author(s):

Juan Sendon-Lago ◽

Lorena Garcia-del Rio ◽

Noemi Eiro ◽

Patricia Diaz-Rodriguez ◽

Leandro Avila ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Body Weight ◽

Conditioned Medium ◽

Local Treatment ◽

Histopathological Analysis ◽

Mrna Levels ◽

Acute Colitis ◽

Tnf Α ◽

Ifn Γ

Inflammatory bowel disease (IBD), including Crohn’s disease (CD) and ulcerative colitis (UC), is increasingly prevalent and current therapies are not completely effective. Mesenchymal stem cells are emerging as a promising therapeutic option. Here, the effect of local hydrogel application loaded with conditioned medium (CM) from human uterine cervical stem cells (hUCESC-CM) in an experimental acute colitis mice model has been evaluated. Colitis induction was carried out in C57BL/6 mice by dissolving dextran sulfate sodium (DSS) in drinking water for nine days. Ulcers were treated by rectal administration of either mesalazine (as positive control) or a mucoadhesive and thermosensitive hydrogel loaded with hUCESC-CM (H-hUCESC-CM). Body weight changes, colon length, and histopathological analysis were evaluated. In addition, pro-inflammatory TNF-α, IL-6, and IFN-γ mRNA levels were measured by qPCR. Treatment with H-hUCESC-CM inhibited body weight loss and colon shortening and induced a significant decrease in colon mucosa degeneration, as well as TNF-α, IFN-γ, and IL-6 mRNA levels. Results indicate that H-hUCESC-CM effectively alleviated DSS-induced colitis in mice, suggesting that H-hUCESC-CM may represent an attractive cell-free therapy for local treatment of IBD.

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