PPAR-γ Is Selectively Upregulated in Caco-2 Cells by Butyrate

doi:10.1006/bbrc.2000.2793

Biochemical and Biophysical Research Communications

Volume 272, Issue 2, 7 June 2000, Pages 380-385

https://doi.org/10.1006/bbrc.2000.2793 Get rights and content

Abstract

The peroxisome proliferator-activated receptor (PPAR)-γ is a nuclear lipid-activable receptor controlling the expression of genes involved in lipid metabolism and adipocyte differentiation. In order to investigate the possible role of PPAR-γ in the differentiation of intestinal epithelial cells, we examined its expression in the human colon carcinoma cell line Caco-2, which undergoes rapid cell differentiation in the presence of butyrate. PPARs were quantified on mRNA level by RT competitive multiplex PCR, the corresponding proteins were determined by Western blot. In contrast to PPAR-α and PPAR-β, PPAR-γ mRNA and protein increased significantly in butyrate-treated Caco-2 cells in a dose- and time-dependent manner. This effect was butyrate-specific, since no change in PPAR-γ expression could be observed after incubation with propionate or valerate. Activation of PPAR-γ by ciglitazone further increased butyrate-induced cell differentiation dose-dependently. These data demonstrate a role for PPAR-γ in the regulation of cell differentiation in Caco-2 cells.

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An important function of the gut microbiome is the fermentation of non-digestible dietary fibers into short chain fatty acids (SCFAs). The three primary SCFAs: acetate, propionate, and butyrate, are key mediators of metabolism and immune cell function in the gut mucosa. We previously demonstrated that butyrate at high concentrations decreased human gut lamina propria (LP) CD4 T cell activation in response to enteric bacteria exposure in vitro. However, to date, the mechanism by which butyrate alters human gut LP CD4 T cell activation remains unknown. In this current study, we sought to better understand how exposure to SCFAs across a concentration range impacted human gut LP CD4 T cell function and activation. LP CD4 T cells were directly activated with T cell receptor (TCR) beads in vitro in the presence of a physiologic concentration range of each of the primary SCFAs. Exposure to butyrate potently inhibited CD4 T cell activation, proliferation, and cytokine (IFNγ, IL-17) production in a concentration dependent manner. Butyrate decreased the proliferation and cytokine production of T helper (Th) 1, Th17 and Th22 cells, with differences noted in the sensitivity of LP versus peripheral blood Th cells to butyrate’s effects. Higher concentrations of propionate and acetate relative to butyrate were required to inhibit CD4 T cell activation and proliferation. Butyrate directly increased the acetylation of both unstimulated and TCR-stimulated CD4 T cells, and apicidin, a Class I histone deacetylase inhibitor, phenocopied butyrate’s effects on CD4 T cell proliferation and activation. GPR43 agonism phenocopied butyrate’s effect on CD4 T cell proliferation whereas a GPR109a agonist did not. Our findings indicate that butyrate decreases in vitro human gut LP CD4 T cell activation, proliferation, and inflammatory cytokine production more potently than other SCFAs, likely through butyrate’s ability to increase histone acetylation, and potentially via signaling through GPR43. These findings have relevance in furthering our understanding of how perturbations of the gut microbiome alter local immune responses in the gut mucosa.
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Citation Excerpt :
PPARs are a group of nuclear receptors and play important roles in lipid metabolism, angiogenesis, immune responses, cell apoptosis, proliferation and differentiation [44]. PPARγ has been suggested in some studies as the main intracellular target of butyrate [45,46]. Activation of PPARγ by butyrate has been demonstrated to maintain gut microbial homeostasis [46], initiate innate immunity against tumour cells[47], and modulate metabolic disorders [14].
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Citation Excerpt :
Our laboratory previous study indicated that dietary pantothenic acid could raise LZ and ACP activities, and down-regulate IFN-γ2 mRNA level in the intestine of grass carp [17]. In human intestinal epithelium cells, butyrate up-regulates peroxisome proliferator-activated receptors γ (PPAR-γ) mRNA level [18], which could increase antimicrobial peptide β-defensins gene expression [19]. Moreover, butyrate could suppress leptin gene expression in bovine mammary epithelial cells [20].
The present study evaluated the effect of dietary sodium butyrate (SB) supplementation on the growth and immune function in the proximal intestine (PI), middle intestine (MI) and distal intestine (DI) of young grass carp (Ctenopharyngodon idella). The fish were fed one powdery sodium butyrate (PSB) diet (1000.0 mg kg⁻¹ diet) and five graded levels of microencapsulated sodium butyrate (MSB) diets: 0.0 (control), 500.0, 1000.0, 1500.0 and 2000.0 mg kg⁻¹ diet for 60 days. Subsequently, a challenge test was conducted by injection of Aeromonas hydrophila. The results indicated that optimal SB supplementation improved the fish growth performance (percent weight gain, specific growth rate, feed intake and feed efficiency) and intestinal growth and function (intestine weight, intestine length, intestinal somatic index, folds height, trypsin, chymotrypsin, lipase and amylase activities), increased beneficial bacteria lactobacillus amount and butyrate concentration, decreased baneful bacteria Aeromonas and Escherichia coli amounts, reduced acetate and propionate concentrations, elevated lysozyme and acid phosphatase activities, increased complement (C3 and C4) and immunoglobulin M contents, and up-regulated β-defensin-1 (rather than DI), hepcidin, liver expressed antimicrobial peptide 2B (LEAP-2B) (except LEAP-2A), Mucin2, interleukin 10 (IL-10), IL-11 (rather than PI), transforming growth factor β1 (rather than PI), transforming growth factor β2 (rather than PI), IL-4/13A, IL-4/13B and inhibitor of κBα (IκBα) mRNA levels, whereas it down-regulated tumor necrosis factor α, interferon γ2, IL-1β (rather than PI), IL-6, IL-8, IL-15 (rather than PI), IL-17D (rather than PI), IL-12p35, IL-12p40 (rather than PI or MI), nuclear factor kappa B p65 (NF-κB p65) (except NF-κB p52), c-Rel (rather than PI or MI), IκB kinase β (IKKβ) (rather than PI), IKKγ (except IKKα), p38 mitogen-activated protein kinase (p38MAPK) and MAPK kinase 6 mRNA levels in three intestinal segments of young grass carp (P < 0.05), suggesting that SB supplementation improves growth and intestinal immune function of fish. Furthermore, according to the positive effect, MSB was superior to PSB on improving growth and enhancing intestinal immune function of fish, and based on feed efficiency of young grass carp, the efficacy of MSB was 3.5-fold higher than that of PSB. Finally, based on percent weight gain, protecting fish against enteritis morbidity and lysozyme activity, the optimal SB supplementation (MSB as SB source) of young grass carp were estimated to be 160.8, 339.9 and 316.2 mg kg⁻¹ diet, respectively.
Butyrate influences intracellular levels of adenine and adenine derivatives in the fungus Penicillium restrictum
2017, Microbiological Research
Citation Excerpt :
Butyrate, a small fatty acid, has received scientific attention due to its important function in the colon of ruminants and mammalians. Butyrate has a role in i) the inhibition of inflammation in the small intestine (Wachtershauser and Stein, 2000), ii) as energy source for colonocytes (Donohoe et al., 2011), iii) the mucosal defence layer by increasing the expression of the MUC2 gene and stimulating the mucin synthesis (Hamer et al., 2008), iv) growth regulation of enterocytes (Hodin et al., 1996; Wachtershauser et al., 2000a,b) and v) cell proliferation and apoptosis of colon and cancer cells (Archer et al., 1998a; Archer et al., 1998b). Furthermore it has been shown that butyrate has an antimicrobial activity against a range of organisms including nematodes, phytopathogenic fungi, yeast and bacteria (Browning et al., 2006; Nguyen et al., 2011).
Butyrate, a small fatty acid, has an important role in the colon of ruminants and mammalians including the inhibition of inflammation and the regulation of cell proliferation. There is also growing evidence that butyrate is influencing the histone structure in mammalian cells by inhibition of histone deacetylation. Butyrate shows furthermore an antimicrobial activity against fungi, yeast and bacteria, which is linked to its toxicity at a high concentration. In fungi there are indications that butyrate induces the production of secondary metabolites potentially via inhibition of histone deacetylases. However, information about the influence of butyrate on growth, primary metabolite production and metabolism, besides lipid catabolism, in fungi is scarce. We have identified the filamentous fungus Penicillium (P.) restrictum as a susceptible target for butyrate treatment in an antimicrobial activity screen. The antimicrobial activity was detected only in the mycelium of the butyrate treated culture. We investigated the effect of butyrate ranging from low (0.001 mM) to high (30 mM), potentially toxic, concentrations on biomass and antimicrobial activity. Butyrate at high concentrations (3 and 30 mM) significantly reduced the fungal biomass. In contrast P. restrictum treated with 0.03 mM of butyrate showed the highest antimicrobial activity. We isolated three antimicrobial active compounds, active against Staphylococcus aureus, from P. restrictum cellular extracts treated with butyrate: adenine, its derivate hypoxanthine and the nucleoside derivate adenosine. Production of all three compounds was increased at low butyrate concentrations. Furthermore we found that butyrate influences the intracellular level of the adenine nucleoside derivate cAMP, an important signalling molecule in fungi and various organisms.
In conclusion butyrate treatment increases the intracellular levels of adenine and its respective derivatives.

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^☆: Abbreviations used: AP, alkaline phosphatase; HETE, hydroxyeicosatetraenoic acid; HODE, hydroxyoctadecadienoic acid; NSAIDs, nonsteroidal anti-inflammatory drugs; 15-d-PGJ₂, 15-desoxy-12,14-prostaglandin J₂; PPAR, peroxisome proliferator-activated receptor; PPRE, peroxisome proliferator responsive element; RXR, retinoid X receptor.

^☆☆: Dedicated to Wolfgang F. Caspary on the occasion of his 60th birthday.

^★: This work was supported by a grant from graduate scholarship, J. W. Goethe University, Frankfurt/Main.

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Regular ArticlePPAR-γ Is Selectively Upregulated in Caco-2 Cells by Butyrate☆,☆☆,★

Abstract

Curr. Opin. Biotech.

FEBS Lett.

Am. J. Clin. Nutr.

Cell

Cell

Cell

J. Biol. Chem.

Gastroenterology

J. Biol. Chem.

Mol. Cell.

Surgery

Biochem. Biophys. Res. Commun.

J. Biol. Chem.

Arch. Biochem. Biophys.

Cell

Regular Article
PPAR-γ Is Selectively Upregulated in Caco-2 Cells by Butyrate☆,☆☆,★