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Ellagitannins from Rubus berries for the control of gastric inflammation: in vitro and in vivo studies.

Sangiovanni E, Vrhovsek U, Rossoni G, Colombo E, Brunelli C, Brembati L, Trivulzio S, Gasperotti M, Mattivi F, Bosisio E, Dell'Agli M - PLoS ONE (2013)

Bottom Line: In vivo the protective effect of ellagitannins was evaluated in a rat model of ethanol-induced gastric lesions.Sanguiin H-6 and lambertianin C, the major ETs present in the extracts, were found to be responsible, at least in part, for the effect of the mixtures.The effect of ETs on CINC-1 was associated to a decrease of NF-κB nuclear translocation in ETs treated animals.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacological and Biomolecular Sciences, Università degli Studi di Milano, Via Balzaretti, Milano, Italy.

ABSTRACT
Ellagitannins have shown anti-inflammatory and anti-Helicobacter pylori properties; however, their anti-inflammatory activity at gastric level was not previously investigated. The aim of this research was to evaluate the effects of ellagitannins from Rubus berries on gastric inflammation. Ellagitannin enriched extracts (ETs) were prepared from Rubus fruticosus L. (blackberry) and Rubus idaeus L. (raspberry). The anti-inflammatory activity was tested on gastric cell line AGS stimulated by TNF-α and IL-1β for evaluating the effect on NF-kB driven transcription, nuclear translocation and IL-8 secretion. In vivo the protective effect of ellagitannins was evaluated in a rat model of ethanol-induced gastric lesions. Rats were treated orally for ten days with 20 mg/kg/day of ETs, and ethanol was given one hour before the sacrifice. Gastric mucosa was isolated and used for the determination of IL-8 release, NF-kB nuclear translocation, Trolox equivalents, superoxide dismutase and catalase activities. In vitro, ETs inhibited TNF-α induced NF-kB driven transcription (IC₅₀: 0.67-1.73 µg/mL) and reduced TNF-α-induced NF-kB nuclear translocation (57%-67% at 2 µg/mL). ETs inhibited IL-8 secretion induced by TNF-α and IL-1β at low concentrations (IC₅₀ range of 0.7-4 µg/mL). Sanguiin H-6 and lambertianin C, the major ETs present in the extracts, were found to be responsible, at least in part, for the effect of the mixtures. ETs of blackberry and raspberry decreased Ulcer Index by 88% and 75% respectively and protected from the ethanol induced oxidative stress in rats. CINC-1 (the rat homologue of IL-8) secretion in the gastric mucosa was reduced in the animals receiving blackberry and raspberry ETs. The effect of ETs on CINC-1 was associated to a decrease of NF-κB nuclear translocation in ETs treated animals. The results of the present study report for the first time the preventing effect of ETs in gastric inflammation and support for their use in dietary regimens against peptic ulcer.

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Effect of ETs from blackberry and raspberry on CINC-1 (IL-8 homologue) ex vivo.CINC-1 release was evaluated using GRO/CINC-1 (rat) EIA kit which uses a polyclonal antibody to rat GRO/CINC-1 labelled with the enzyme horseradish peroxidase. After a short incubation (10 minutes) the enzyme reaction was followed by spectroscopy (signal read 450 nm, 0.1 s). The concentration of rat GRO/CINC-1 in the samples was determined by interpolation with a GRO/CINC-1 standard curve. The results (mean ± se, n = 6) are expressed as pg of CINC-1 per mL of sample. * p<0.05, ** p<0.01.
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pone-0071762-g009: Effect of ETs from blackberry and raspberry on CINC-1 (IL-8 homologue) ex vivo.CINC-1 release was evaluated using GRO/CINC-1 (rat) EIA kit which uses a polyclonal antibody to rat GRO/CINC-1 labelled with the enzyme horseradish peroxidase. After a short incubation (10 minutes) the enzyme reaction was followed by spectroscopy (signal read 450 nm, 0.1 s). The concentration of rat GRO/CINC-1 in the samples was determined by interpolation with a GRO/CINC-1 standard curve. The results (mean ± se, n = 6) are expressed as pg of CINC-1 per mL of sample. * p<0.05, ** p<0.01.

Mentions: The administration of ethanol caused a higher release of CINC-1 (the rat homologue of human IL-8) from 12.8 pg/ml in the tissue from control animals to 28 pg/ml in ethanol treated rats (Figure 9). In animals treated with ETblack and ETrasp the amount of CINC-1 was significantly lower with respect to ethanol (16.5 ± 1.9 and 22.2 ± 2.3 pg/ml, respectively). In rats treated with quercetin as positive control CINC-1 levels were 15.9 ± 0.72 pg/ml. The effect of ETs on CINC-1 was associated to a decrease of NF-κB translocation in ETblack and ETrasp animals in comparison with control and ethanol treated animals. In the tissue of ETblack and ETrasp animals, NF-κB translocation was inhibited by 38±0.11% (mean ± sd, n = 6, p<0.001) and 72±1.6% (n = 6, p<0.0001) respectively, with respect to ethanol. No difference was observed between control and ethanol group. No data are present in the literature as regards NF-κB nuclear translocation in vivo in the animal model of ethanol-induced ulcer. An explanation is that the damage of the tissue does not allow to properly isolate the nuclear fraction.


Ellagitannins from Rubus berries for the control of gastric inflammation: in vitro and in vivo studies.

Sangiovanni E, Vrhovsek U, Rossoni G, Colombo E, Brunelli C, Brembati L, Trivulzio S, Gasperotti M, Mattivi F, Bosisio E, Dell'Agli M - PLoS ONE (2013)

Effect of ETs from blackberry and raspberry on CINC-1 (IL-8 homologue) ex vivo.CINC-1 release was evaluated using GRO/CINC-1 (rat) EIA kit which uses a polyclonal antibody to rat GRO/CINC-1 labelled with the enzyme horseradish peroxidase. After a short incubation (10 minutes) the enzyme reaction was followed by spectroscopy (signal read 450 nm, 0.1 s). The concentration of rat GRO/CINC-1 in the samples was determined by interpolation with a GRO/CINC-1 standard curve. The results (mean ± se, n = 6) are expressed as pg of CINC-1 per mL of sample. * p<0.05, ** p<0.01.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3733869&req=5

pone-0071762-g009: Effect of ETs from blackberry and raspberry on CINC-1 (IL-8 homologue) ex vivo.CINC-1 release was evaluated using GRO/CINC-1 (rat) EIA kit which uses a polyclonal antibody to rat GRO/CINC-1 labelled with the enzyme horseradish peroxidase. After a short incubation (10 minutes) the enzyme reaction was followed by spectroscopy (signal read 450 nm, 0.1 s). The concentration of rat GRO/CINC-1 in the samples was determined by interpolation with a GRO/CINC-1 standard curve. The results (mean ± se, n = 6) are expressed as pg of CINC-1 per mL of sample. * p<0.05, ** p<0.01.
Mentions: The administration of ethanol caused a higher release of CINC-1 (the rat homologue of human IL-8) from 12.8 pg/ml in the tissue from control animals to 28 pg/ml in ethanol treated rats (Figure 9). In animals treated with ETblack and ETrasp the amount of CINC-1 was significantly lower with respect to ethanol (16.5 ± 1.9 and 22.2 ± 2.3 pg/ml, respectively). In rats treated with quercetin as positive control CINC-1 levels were 15.9 ± 0.72 pg/ml. The effect of ETs on CINC-1 was associated to a decrease of NF-κB translocation in ETblack and ETrasp animals in comparison with control and ethanol treated animals. In the tissue of ETblack and ETrasp animals, NF-κB translocation was inhibited by 38±0.11% (mean ± sd, n = 6, p<0.001) and 72±1.6% (n = 6, p<0.0001) respectively, with respect to ethanol. No difference was observed between control and ethanol group. No data are present in the literature as regards NF-κB nuclear translocation in vivo in the animal model of ethanol-induced ulcer. An explanation is that the damage of the tissue does not allow to properly isolate the nuclear fraction.

Bottom Line: In vivo the protective effect of ellagitannins was evaluated in a rat model of ethanol-induced gastric lesions.Sanguiin H-6 and lambertianin C, the major ETs present in the extracts, were found to be responsible, at least in part, for the effect of the mixtures.The effect of ETs on CINC-1 was associated to a decrease of NF-κB nuclear translocation in ETs treated animals.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacological and Biomolecular Sciences, Università degli Studi di Milano, Via Balzaretti, Milano, Italy.

ABSTRACT
Ellagitannins have shown anti-inflammatory and anti-Helicobacter pylori properties; however, their anti-inflammatory activity at gastric level was not previously investigated. The aim of this research was to evaluate the effects of ellagitannins from Rubus berries on gastric inflammation. Ellagitannin enriched extracts (ETs) were prepared from Rubus fruticosus L. (blackberry) and Rubus idaeus L. (raspberry). The anti-inflammatory activity was tested on gastric cell line AGS stimulated by TNF-α and IL-1β for evaluating the effect on NF-kB driven transcription, nuclear translocation and IL-8 secretion. In vivo the protective effect of ellagitannins was evaluated in a rat model of ethanol-induced gastric lesions. Rats were treated orally for ten days with 20 mg/kg/day of ETs, and ethanol was given one hour before the sacrifice. Gastric mucosa was isolated and used for the determination of IL-8 release, NF-kB nuclear translocation, Trolox equivalents, superoxide dismutase and catalase activities. In vitro, ETs inhibited TNF-α induced NF-kB driven transcription (IC₅₀: 0.67-1.73 µg/mL) and reduced TNF-α-induced NF-kB nuclear translocation (57%-67% at 2 µg/mL). ETs inhibited IL-8 secretion induced by TNF-α and IL-1β at low concentrations (IC₅₀ range of 0.7-4 µg/mL). Sanguiin H-6 and lambertianin C, the major ETs present in the extracts, were found to be responsible, at least in part, for the effect of the mixtures. ETs of blackberry and raspberry decreased Ulcer Index by 88% and 75% respectively and protected from the ethanol induced oxidative stress in rats. CINC-1 (the rat homologue of IL-8) secretion in the gastric mucosa was reduced in the animals receiving blackberry and raspberry ETs. The effect of ETs on CINC-1 was associated to a decrease of NF-κB nuclear translocation in ETs treated animals. The results of the present study report for the first time the preventing effect of ETs in gastric inflammation and support for their use in dietary regimens against peptic ulcer.

Show MeSH
Related in: MedlinePlus