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Cardiovascular Health & Anti-inflammatory Benefits: In-Vitro

Displaying 1 - 10 of 38

Inhibitory Effect of Cranberry Extract on LPS Induced Inflammatory Response in RAW246.7 Cells

Posted: 
February 19, 2019
Authors: 
Gao NY, Zhao YM, Liu DL, Sun HG, Gao XX
Journal: 
Food Research and Development; 2018. 39(16):1-7.
Abstract: 

To study the anti-inflammatory effect of cranberry extract on inflammation suppression induced by lipopolysaccharide, and explore its mechanism. Cell inflammatory model was established with RAW264.7 cells treated with lipopolysaccharide. Cell viability of RAW264.7 cells treated with cranberry extract were analyzed by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. The effect of cranberry extract on nucleus was observed by 4',6-diamidino-2-phenylindole(DAPI)staining. The activity of nitric oxide synthase (NOS) was determined by fluorescence analysis. Enzyme-linked immunosorbent assay (ELISA) for determination of IL-1 beta , IL-6 and TNF- alpha . RAW264.7 cells were treated with cranberry extract for 24 h, and the expression of Keap1, Nrf2, HO-1, IKK alpha / beta and NF- kappa Bp65 were detected by Western blotting. The result showed that the inflammatory model was established by 5 micro g/mL lipopolysaccharide, the highest level of inflammation was reached at 24 hours. There was no significant toxic effect on RAW246.7 cells in the range of 5 micro g/mL-400 micro g/mL, and the cell nucleus was intact and without obvious damage. Compared with the model group, cranberry extract could significantly inhibit the activity of NOS and decreased the content of IL-1 beta , IL-6, TNF- alpha with the increase of dose. The Western blot result showed that cranberry extract inhibited the expression of Keap1, IKK alpha / beta , NF- kappa Bp65 and increase the expression of Nrf2 and HO-1 protein levels. These results suggest that cranberry extract can inhibit the inflammatory response induced by lipopolysaccharide, and its mechanism may be related to activation of Keap1/Nrf2/HO-1 signaling pathway and NF- kappa Bp65.

Atomic force microscopy-guided fractionation reveals the influence of cranberry phytochemicals on adhesion of Escherichia coli.

Posted: 
August 22, 2016
Authors: 
Gupta P; Song B; Neto C; Camesano TA.
Journal: 
Food & Function. 7(6):2655-66, 2016
Abstract: 

Cranberry juice has been long used to prevent infections because of its effect on the adhesion of the bacteria to the host surface. Proanthocyanidins (PACs) comprise of one of the major classes of phytochemicals found in cranberry, which have been extensively studied and found effective in combating adhesion of pathogenic bacteria. The role of other cranberry constituents in impacting bacterial adhesion haven't been studied very well. In this study, cranberry juice fractions were prepared, characterized and tested for their effect on the surface adhesion of the pathogenic clinical bacterial strain E. coli B78 and non-pathogenic control E. coli HB101. The preparations tested included crude cranberry juice extract (CCE); three fractions containing flavonoid classes including proanthocyanidins, anthocyanins and flavonols; selected sub-fractions, and commercially available flavonol glycoside, quercetin-3-O-galactoside. Atomic force microscopy (AFM) was used to quantify the adhesion forces between the bacterial surface and the AFM probe after the treatment with the cranberry fractions. Adhesion forces of the non-pathogenic, non fimbriated lab strain HB101 are small (average force 0.19 nN) and do not change with cranberry treatments, whereas the adhesion forces of the pathogenic, Dr adhesion E. coli strain B78 (average force of 0.42 nN) show a significant decrease when treated with cranberry juice extract or fractions (average force of 0.31 nN, 0.37 nN and 0.39 nN with CCE, Fraction 7 and Fraction 4 respectively). In particular, the fractions that contained flavonols in addition to PACs were more efficient at lowering the force of adhesion (average force of 0.31 nN-0.18 nN between different sub-fractions containing flavonols and PACs). The sub-fractions containing flavonol glycosides (from juice, fruit and commercial quercetin) all resulted in reduced adhesion of the pathogenic bacteria to the model probe. This strongly suggests the anti adhesive role of other classes of cranberry compounds in conjunction with already known PACs and may have implications for development of alternative anti bacterial treatments.

Deleterious effect of p-cresol on human colonic epithelial cells prevented by proanthocyanidin-containing polyphenol extracts from fruits and proanthocyanidin bacterial metabolites.

Posted: 
August 22, 2016
Authors: 
Wong X; Carrasco-Pozo C; Escobar E; Navarrete P; Blachier F; Andriamihaja M; Lan A; Tome D; Cires MJ; Pastene E; Gotteland M.
Journal: 
Journal of Agricultural and Food Chemistry; 2016. 64(18):3574-3583
Abstract: 

The protective effect of proanthocyanidin-containing polyphenol extracts from apples, avocados, cranberries, grapes, or proanthocyanidin microbial metabolites was evaluated in colonic epithelial cells exposed to p-cresol, a deleterious compound produced by the colonic microbiota from L-tyrosine. In HT29 Glc-/+ cells, p-cresol significantly increased LDH leakage and decreased ATP contents, whereas in Caco-2 cell monolayers, it significantly decreased the transepithelial electrical resistance and increased the paracellular transport of FITC-dextran. The alterations induced by p-cresol in HT29 Glc-/+ cells were prevented by the extracts from cranberries and avocados, whereas they became worse by extracts from apples and grapes. The proanthocyanidin bacterial metabolites decreased LDH leakage, ameliorating cell viability without improving intracellular ATP. All of the polyphenol extracts and proanthocyanidin bacterial metabolites prevented the p-cresol-induced alterations of barrier function. These results suggest that proanthocyanidin-containing polyphenol extracts and proanthocyanidin metabolites likely contribute to the protection of the colonic mucosa against the deleterious effects of p-cresol.

Cranberry (Vaccinium macrocarpon) Oligosaccharides Decrease Biofilm Formation by Uropathogenic Escherichia Coli.

Posted: 
March 23, 2016
Authors: 
Sun J, Marais JP, Khoo C, LaPlante K, Vejborg RM, Givskov M, Tolker-Nielsen T, Seeram NP, Rowley DC
Journal: 
J Funct Food 17:235-242
Abstract: 

The preventive effects of the American cranberry (Vaccinium macrocarpon) against urinary tract infections are supported by extensive studies which have primarily focused on its phenolic constituents. Herein, a phenolic-free carbohydrate fraction (designated cranf1b-F2) was purified from cranberry fruit using ion exchange and size exclusion chromatography. MALDI-TOF-MS analysis revealed that the cranf1b-F2 constituents are predominantly oligosaccharides possessing various degrees of polymerisation and further structural analysis (by GC-MS and NMR) revealed mainly xyloglucan and arabinan residues. In antimicrobial assays, cranf1b-F2 (at 1.25 mg/mL concentration) reduced biofilm production by the uropathogenic Escherichia coli CFT073 strain by over 50% but did not inhibit bacterial growth. Cranf1b-F2 (ranging from 0.625 to 10 mg/mL) also inhibited biofilm formation of the non-pathogenic E. coli MG1655 strain up to 60% in a concentration-dependent manner. These results suggest that cranberry oligosaccharides, in addition to its phenolic constituents, may play a role in its preventive effects against urinary tract infections.

Fluorescent Labeling of Cranberry Proanthocyanidins with 5-([4,6-dichlorotriazin-2-yl]amino)Fluorescein (DTAF)

Posted: 
March 23, 2016
Authors: 
Feliciano RP, Heintz JA, Krueger CG, Vestling MM, Reed JD
Journal: 
Food Chem 166:337-45
Abstract: 

A novel methodology was developed to elucidate proanthocyanidins (PAC) interaction with extra-intestinal pathogenic Escherichia coli (ExPEC). PAC inhibit ExPEC invasion of epithelial cells and, therefore, may prevent transient gut colonization, conferring protection against subsequent extra-intestinal infections, such as urinary tract infections. Until now PAC have not been chemically labeled with fluorophores. In this work, cranberry PAC were labeled with 5-([4,6-dichlorotriazin-2-yl]amino) fluorescein (DTAF), detected by high-performance liquid chromatography with diode-array detection and characterized by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). We report single and double fluorescent-labeled PAC with one or two chlorine atoms displaced from DTAF in alkaline pH via nucleophilic substitution. Fluorescent labeling was confirmed by fragmentation experiments using MALDI-TOF/TOF MS. Fluorescent labeled PAC were able to promote ExPEC agglutination when observed with fluorescence microscopy. DTAF tagged PAC may be used to trace the fate of PAC after they agglutinate ExPEC and follow PAC-ExPEC complexes in cell culture assays.

New Functionally-Enhanced Soy Proteins as Food Ingredients with Anti-Viral Activity.

Posted: 
March 23, 2016
Authors: 
Turmagambetova AS, Sokolova NS, Bogoyavlenskiy AP, Berezin VE, Lila MA, Cheng DM, Dushenkov V
Journal: 
VirusDis 26(3):123-32
Abstract: 

Respiratory viruses are a major public health problem because of their prevalence and high morbidity rate leading to considerable social and economic implications. Cranberry has therapeutic potential attributed to a comprehensive list of phytochemicals including anthocyanins, flavonols, and unique A-type proanthocyanidins. Soy flavonoids, including isoflavones, have demonstrated anti-viral effects in vitro and in vivo. Recently, it was demonstrated that edible proteins can efficiently sorb and concentrate cranberry polyphenols, including anthocyanins and proanthocyanins, providing greatly stabilized matrices suitable for food products. The combination of cranberry and soy phytoactives may be an effective dietary anti-viral resource. Anti-viral properties of both cranberry juice-enriched and cranberry pomace polyphenol-enriched soy protein isolate (CB-SPI and CBP-SPI) were tested against influenza viruses (H7N1, H5N3, H3N2), Newcastle disease virus and Sendai virus in vitro and in ovo. In our experiments, preincubation with CB-SPI or CBP-SPI resulted in inhibition of virus adsorption to chicken red blood cells and reduction in virus nucleic acid content up to 16-fold, however, CB-SPI and CBP-SPI did not affect hemagglutination. Additionally, CB-SPI and CBP-SPI inhibited viral replication and infectivity more effectively than the commercially available anti-viral drug Amizon. Results suggest CB-SPI and CBP-SPI may have preventative and therapeutic potential against viral infections that cause diseases of the respiratory and gastro-intestinal tract.

Comparative in vitro fermentations of cranberry and grape seed polyphenols with colonic microbiota.

Posted: 
September 28, 2015
Authors: 
Sanchez-Patan F, Barroso E, van de Wiele T, Jimenez-Giron A, Martin-Alvarez PJ, Moreno-Arribas MV, Martinez-Cuesta MC, Pelaez C, Requena T, Bartolome B
Journal: 
Food Chem 183:273-82,
Abstract: 

In this study, we have assessed the phenolic metabolism of a cranberry extract by microbiota obtained from the ascending colon and descending colon compartments of a dynamic gastrointestinal simulator (SHIME). For comparison, parallel fermentations with a grape seed extract were carried out. Extracts were used directly without previous intestinal digestion. Among the 60 phenolic compounds targeted, our results confirmed the formation of phenylacetic, phenylpropionic and benzoic acids as well as phenols such as catechol and its derivatives from the action of colonic microbiota on cranberry polyphenols. Benzoic acid (38.4mug/ml), 4-hydroxy-5-(3'-hydroxyphenyl)-valeric acid (26.2mug/ml) and phenylacetic acid (19.5mug/ml) reached the highest concentrations. Under the same conditions, microbial degradation of grape seed polyphenols took place to a lesser extent compared to cranberry polyphenols, which was consistent with the more pronounced antimicrobial effect observed for the grape seed polyphenols, particularly against Bacteroides, Prevotella and Blautia coccoides-Eubacterium rectale.

Effects of Proanthocyanidins on Adhesion, Growth, and Virulence of Highly Virulent Extraintestinal Pathogenic Escherichia coli Argue for Its Use to Treat Oropharyngeal Colonization and Prevent Ventilator-Associated Pneumonia.

Posted: 
September 28, 2015
Authors: 
Margetis D, Roux D, Gaudry S, Messika J, Bouvet O, Branger C, Ponnuswamy P, Oufella HA, Dreyfuss D, Denamur E, Ricard JD
Journal: 
Crit Care Med 43(6):e170-8,
Abstract: 

OBJECTIVE: In the context of increasing microbial resistance and limited new antimicrobials, we aimed to study the antimicrobial effects of cranberry proanthocyanidin extracts on Escherichia coli growth, adhesion to epithelial cells, and lung infection.
DESIGN: Experimental in vitro and in vivo investigation.
SETTING: University research laboratory.
SUBJECTS: Seventy-eight 6- to 8-week-old male Balb/C mice.
INTERVENTIONS: In vitro, the effect of increasing concentrations of cranberry proanthocyanidin on bacterial growth of different clinical E. coli isolates was evaluated. Ex vivo, adhesion of E. coli to fresh human buccal epithelial cells was measured in the presence or absence of cranberry proanthocyanidin using microscopy. In vivo, lung bacterial count, pulmonary immune response (neutrophil murine chemokine keratinocyte-derived cytokine measurement and polymorphonuclear recruitment in bronchoalveolar lavage fluid), and lethality were evaluated in a pneumonia mouse model with E. coli precultured with or without cranberry proanthocyanidin. E. coli isolates originated from ventilated ICU patients with respiratory tract colonization or ventilator- associated pneumonia. They differed in number of virulence genes.
MEASUREMENTS AND MAIN RESULTS: A significant inhibition of bacterial growth was observed with increasing concentration of cranberry proanthocyanidin, affecting both time to maximal growth and maximal growth rate (p CONCLUSION: Cranberry proanthocyanidins exhibit potent effects on growth, adhesion, and virulence of oropharyngeal and lung isolates of E. coli, suggesting that cranberry proanthocyanidin could be of clinical interest to reduce oropharyngeal colonization and prevent lung infection.

The cranberry flavonoids PAC DP-9 and quercetin aglycone induce cytotoxicity and cell cycle arrest and increase cisplatin sensitivity in ovarian cancer cells.

Posted: 
September 28, 2015
Authors: 
Wang Y, Han A, Chen E, Singh RK, Chichester CO, Moore RG, Singh AP, Vorsa N
Journal: 
Int J Oncol 46(5):1924-34
Abstract: 

Cranberry flavonoids (flavonols and flavan-3-ols), in addition to their antioxidant properties, have been shown to possess potential in vitro activity against several cancers. However, the difficulty of isolating cranberry compounds has largely limited anticancer research to crude fractions without well-defined compound composition. In this study, individual cranberry flavonoids were isolated to the highest purity achieved so far using gravity and high performance column chromatography and LC-MS characterization. MTS assay indicated differential cell viability reduction of SKOV-3 and OVCAR-8 ovarian cancer cells treated with individual cranberry flavonoids. Treatment with quercetin aglycone and PAC DP-9, which exhibited the strongest activity, induced apoptosis, led to caspase-3 activation and PARP deactivation, and increased sensitivity to cisplatin. Furthermore, immunofluorescence microscopy and western blot study revealed reduced expression and activation of epidermal growth factor receptor (EGFR) in PAC DP-9 treated SKOV-3 cells. In addition, quercetin aglycone and PAC DP-9 deactivated MAPK-ERK pathway, induced downregulation of cyclin D1, DNA-PK, phospho-histone H3 and upregulation of p21, and arrested cell cycle progression. Overall, this study demonstrates promising in vitro cytotoxic and anti-proliferative properties of two newly characterized cranberry flavonoids, quercetin aglycone and PAC DP-9, against ovarian cancer cells.

Inhibition of interleukin 1beta-stimulated interleukin-6 production by cranberry components in human gingival epithelial cells: effects on nuclear factor B and activator protein 1 activation pathways

Posted: 
July 25, 2014
Authors: 
Tipton DA, Carter TB, Dabbous MKh
Journal: 
J Periodontal Res 49(4):437-47
Abstract: 

BACKGROUND AND OBJECTIVE: In periodontitis, gingival epithelial cells can produce interleukin (IL)-6, a regulator of osteoclastic bone resorption, in response to IL-1beta. IL-1beta regulates cytokine expression via signaling pathways, including nuclear factor (NF)-B and mitogen activated protein kinase (MAPK)/activator protein (AP)-1. Cranberry proanthocyanidins (PACs) inhibit IL-1beta-stimulated IL-6 production, but specific mechanisms are unclear. The objectives of this study were to determine effects of cranberry PACs on NF-B and MAPK/AP-1 activation of IL-1beta-stimulated IL-6 production in gingival epithelial cells.

MATERIAL AND METHODS: Cranberry high molecular weight non-dialyzable material (NDM), rich in PACs, was derived from cranberry juice. Human gingival epithelial cells [Smulow-Glickman (S-G)] were incubated with IL-1beta in the presence or absence of NDM or inhibitors of NF-B, [nemo-binding domain (NBD) peptide] or AP-1 (SP600125), and IL-6 levels were measured by ELISA. Effects of NDM on IL-1beta-activated NF-B and AP-1 and phosphorylated intermediates in both pathways were measured in cell extracts via binding to specific oligonucleotides and specific sandwich ELISAs, respectively. Data were analyzed using ANOVA and Scheffe's F procedure for post hoc comparisons.

RESULTS: IL-1beta (> 0.1 nm) caused a time- and dose-dependent stimulation of S-G epithelial cell IL-6 production (p

CONCLUSION: In S-G epithelial cells, IL-1beta appeared to upregulate IL-6 production via activation of both NF-B and MAPK/AP-1 signaling pathways because cranberry NDM decreased nuclear levels of IL-1beta-activated NF-B (p65) and AP-1 (phospho-c-Jun) and strongly inhibited IL-6 production. Lack of inhibition of phosphorylation of IBalpha, c-Jun or ERK1/2 suggested that NDM might affect both pathways downstream from those points in S-G cells, such as ubiquitination and proteosomal degradation of IBalpha, or inhibition of nuclear activity of c-Jun and/or ERK1/2. Defining these points of inhibition precisely may help identify molecular targets of cranberry polyphenols. 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

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