Aleksandra S. Kristo

Wild blueberry (Vaccinium angustifolium) consumption improves inflammatory status in the obese Zucker rat model of the metabolic syndrome

The metabolic syndrome (MetS) is a major public health problem in the United States. Chronic inflammation is a critical component of the MetS, leading to dramatically increased risk of type II diabetes and cardiovascular disease. This study investigates the ability of a wild-blueberry-enriched diet to improve the proinflammatory status associated with MetS in the obese Zucker rat (OZR). Circulating levels of key inflammatory markers and their expression in the liver and abdominal adipose tissue were examined in OZR and its genetic control, the lean Zucker rat (LZR), after feeding a control or an 8% wild blueberry diet (WB) for 8 weeks from age 8 to 16 weeks. In the OZR, WB consumption resulted in decreased plasma concentrations of tumor necrosis factor (TNF)-α (-25.6%, P<.05), interleukin (IL)-6 (-14.9%, P<.05) and C-reactive protein (CRP) (-13.1%, P<.05) and increased adiponectin concentration (+21.8%, P<.05). Furthermore, expression of IL-6, TNF-α and nuclear factor (NF)-kB was down-regulated in both the liver (-65%, -59% and -25%, respectively) and the abdominal adipose tissue (-64%, -52% and -65%), while CRP expression was down-regulated only in the liver (-25%). In the abdominal adipose tissue, similar trends were also observed in LZR following WB treatment, with decreased liver expression of NF-kB, CRP, IL-6 and TNF-α (-24%, -16%, -21% and -50%) and increased adiponectin expression (+25%). Results of this study suggest that wild blueberry consumption exerts an overall anti-inflammatory effect in the OZR, a model of the metabolic syndrome.

Dietary enrichment with wild blueberries (Vaccinium angustifolium) affects the vascular reactivity in the aorta of young spontaneously hypertensive rats.

We have previously reported on the positive effects of wild blueberries on arterial contractile response to alpha(1) adrenergic stimuli and on endothelium-mediated vasorelaxation. Our present study was designed to evaluate the effects of the dietary enrichment with wild blueberries on aortic function and reactivity in the developmental phase of essential hypertension in spontaneously hypertensive rats (SHR). We investigated the possible influence blueberries may have on the acetylcholine (Ach)-induced endothelium-dependent vasorelaxation and phenylephrine-induced vasoconstriction in young SHRs, as well as the contribution of the nitric oxide (NO) synthase and cyclooxygenase (COX) pathways in each of the above responses in an animal model with dysfunctional endothelium. Vascular ring studies were conducted in 3-mm isolated rat aortic ring preparations to investigate vasoconstriction induced by l-Phenylephrine (Phe, 10(-8) to 3x10(-6)M) and vasorelaxation induced by acetylcholine (Ach, 10(-9) to 3x10(-6)M). The major findings of our study were that in Phe-induced vasoconstriction, SHR-BB aortas relaxed to a greater degree in comparison to controls when mefenamic acid (MFA) was present and that the incubation with this COX inhibitor failed to restore - and in fact decreased - the maximum vasodilator response to Ach, in comparison to controls. Our vessel reactivity index (pD(2)) observations indicate that blueberries appear to modulate cell membrane-agonist (Ach) interactions primarily in response to Ach in the young SHR model, but not to the alpha(1) adrenoreceptor agonist. Incorporating wild blueberries in the diet seems to affect the endothelium-dependent vasorelaxation by modulating alternative metabolic pathway(s) (such as affecting the production/activity of COX-derived products) in the young SHR aorta.

A wild blueberry-enriched diet (Vaccinium angustifolium) improves vascular tone in the adult spontaneously hypertensive rat.

The effect of a wild blueberry-enriched diet on vasoconstriction and vasorelaxation was examined in the adult, 20-week-old spontaneously hypertensive rat (SHR) after 8 weeks of a control (C) or an 8% wild blueberry (WB) diet. Nitric oxide (NO)- and cyclooxygenase (COX)-mediated aortic responses were examined ex vivo with the agonists L-phenylephrine (Phe) and acetylcholine (Ach), in the absence or presence of the NO synthase (NOS) inhibitor L-NG-monomethyl arginine (L-NMMA) or the COX inhibitor mefenamic acid (MFA). The vasoconstriction elicited by Phe was reduced in the WB group, attributed to the NO pathway, favoring a lower vascular tone under basal conditions. Acetylcholine-induced vasorelaxation in the WB group was possibly mediated through the COX, but not the NO pathway. These findings document the potential of wild blueberries to modify major pathways of vasomotor control and improve the vascular tone in the adult SHR with endothelial dysfunction.

Lowbush wild blueberries have the potential to modify gut microbiota and xenobiotic metabolism in the rat colon.

The gastrointestinal tract is populated by an array of microbial species that play an important role in metabolic and immune functions. The composition of microorganisms is influenced by the components of the host’s diet and can impact health. In the present study, dietary enrichment of lowbush wild blueberries (LWB) was examined to determine their effect on colon microbial composition and their potential in promoting gut health. The microbial composition and functional potential of the colon microbiota from Sprague Dawley rats fed control diets (AIN93) and LWB-enriched diets (AIN93+8% LWB powder substituting for dextrose) for 6 weeks were assessed using Illumina shotgun sequencing and bioinformatics tools. Our analysis revealed an alteration in the relative abundance of 3 phyla and 22 genera as representing approximately 14 and 8% of all phyla and genera identified, respectively. The LWB-enriched diet resulted in a significant reduction in the relative abundance of the genera Lactobacillus and Enterococcus. In addition, hierarchal analysis revealed a significant increase in the relative abundance of the phylum Actinobacteria, the order Actinomycetales, and several novel genera under the family Bifidobacteriaceae and Coriobacteriaceae, in the LWB group. Functional annotation of the shotgun sequences suggested that approximately 9% of the 4709 Kyoto Encyclopaedia of Gene and Genome (KEGG) hits identified were impacted by the LWB-diet. Open Reading Frames (ORFs) assigned to KEGG category xenobiotics biodegradation and metabolism were significantly greater in the LWB-enriched diet compared to the control and included the pathway for benzoate degradation [PATH:ko00362] and glycosaminoglycan degradation [PATH:ko00531]. Moreover, the number of ORFs assigned to the bacterial invasion of epithelial cells [PATH:ko05100] pathway was approximately 8 fold lower in the LWB group compared to controls. This study demonstrated that LWBs have the potential to promote gut health and can aid in the development of optimal diets.

The temporal effect of a wild blueberry (Vaccinium angustifolium)-enriched diet on vasomotor tone in the Sprague-Dawley rat.

BACKGROUND AND AIMS We have previously reported that wild blueberry (Vaccinium angustifolium)-enriched diets (WB) attenuate aortic adrenergic response through endothelial-mediated pathways. The duration of dietary intervention necessary to induce the positive changes on vasomotor tone has not been studied to date. Thus, our objective was to investigate the temporal effect of WB consumption on vascular function and reactivity in Sprague-Dawley (SD) rat aorta after 4 and 7 weeks of dietary treatment. METHODS AND RESULTS Forty male SD rats were randomly assigned to a control (AIN-93) (C) or a WB diet for 4 or 7 weeks. Vascular ring studies were conducted in 3-mm isolated rat aortic rings to investigate vasoconstriction induced by six doses of the α(1)-adrenergic agonist, L-phenylephrine (Phe, 10(-8)-3×10(-6) M) alone or in the presence of the NOS inhibitor, L-N(G)-monomethyl-arginine (L-NMMA, 10(-4)M). The maximum force of contraction (F(max)) and vessel sensitivity (pD(2)) were determined. Analysis of variance revealed no significant differences on F(max) after 4 weeks of the WB diet but only a significant increase in pD(2) in the absence of L-NMMA. Seven week WB consumption significantly attenuated contraction in response to L-Phe and resulted in lower pD(2). Inhibition of NOS induced a significant increase in the constrictor response in both diet groups at both time periods, with the WB group fed for 7 weeks having the greater response. CONCLUSION Thus wild blueberries incorporated into the diet at 8% w/w positively affect vascular smooth muscle contractility and sensitivity but these effects are evident only after 7 weeks of WB consumption.

Wild blueberries attenuate risk factors of the metabolic syndrome

The ability of a wild blueberry-enriched diet to improve risk factors related to Metabolic Syndrome (MetS) such as endothelial dysfunction and chronic inflammation in the Obese Zucker Rat (OZR), a model of the MetS, was studied. Obese Zucker Rats (OZRs) and their lean controls (LZR) were placed either on a Wild Blueberry-enriched (WB) or a control (C) diet for 8 weeks. Obese Zucker rats exhibited reduced vasoconstrictor response to phenylephrine (Phe) and exaggerated vasorelaxant response to acetylcholine (Ach). WB diet partially restored Phe-induced constrictor responses and attenuated Ach-induced relaxant responses in OZR. Plasma nitric oxide (NO) was significantly attenuated and aortic effluent prostaglandin I2 PGI2 concentration significantly increased in the WB diet. Downregulation of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) expression in the OZR aorta was observed in the WB diet. WB consumption decreased plasma concentrations of tumor necrosis factor (TNF)(–25.6%, P< 0.05), interleukin (IL)-6 (–14.9%, P< 0.05), C-reactive protein (CRP) (–13.1%, P< 0.05) and increased adiponectin concentration (+21.8%, P< 0.05). Expression of IL-6, TNFand nuclear factor (NF)-kB was downregulated in both the liver (–65%, –59% and –25%, respectively) and the abdominal adipose tissue (–64%, –52% and –65%), while CRP expression was down regulated only in the liver (–25%). Thus, WB consumption improved endothelial function and exerted an anti-inflammatory effect in the OZR.

Wild blueberry consumption affects aortic vascular function in the obese Zucker rat.

This study evaluates the effect of wild blueberry (WB) consumption on the biomechanical properties of the aorta in the obese Zucker rat (OZR), a model of the metabolic syndrome. Thirty-six OZRs and 36 lean controls (lean Zucker rats) were placed either on a WB-enriched or a control (C) diet for 8 weeks. Phenylephrine (Phe)-mediated vasoconstriction and acetylcholine (Ach)-mediated vasorelaxation in the aortic vessel were investigated, as well as the contribution of the nitric oxide synthase and cyclooxygenase (COX) pathways in each of the above responses by using specific inhibitors. Obese Zucker rats exhibited a reduced vasocontstrictor response to Phe and an exaggerated vasorelaxant response to Ach. The WB diet partially restored Phe-induced constrictor responses and attenuated Ach-induced relaxant responses in OZR. Plasma nitric oxide was significantly attenuated (22.1 ± 1.1 μmol·L(-1), WB vs 25.6 ± 1.4 μmol·L(-1), C, p ≤ 0.05) with the WB diet. Thromboxane A2 levels in the aortic effluent were not significantly affected in the WB diet group, while PGI2 concentration significantly increased (766.5 ± 92.2 pg·mg(-1) aorta in the WB vs 571.7 ± 37.8 pg·g(-1) aorta in the C group, p ≤ 0.05). Downregulation of inducible nitric oxide synthase and COX2 expression in the OZR aorta was observed in the WB diet group. In conclusion, WB consumption altered the biomechanical properties of the OZR aorta by partially restoring the impaired Phe-induced constrictor responses and attenuating the exaggerated response to Ach-induced vasorelaxation.

Wild blueberry (V. angustifolium)-enriched diets alter aortic glycosaminoglycan profile in the spontaneously hypertensive rat

Glycosaminoglycans (GAGs) are essential polysaccharide components of extracellular matrix and cell surface with key roles on numerous vascular wall functions. Previous studies have documented a role of wild blueberries on the GAG profile of the Sprague-Dawley rat with a functional endothelium as well as in the vascular tone of the spontaneously hypertensive rat (SHR) with endothelial dysfunction. In the present study, the effect of wild blueberries on the composition and structure of aortic GAGs was examined in 20-week-old SHRs after 8 weeks on a control (C) or a wild blueberry-enriched diet (WB). Aortic tissue GAGs were isolated following pronase digestion and anion-exchange chromatography. Treatment of the isolated populations with specific GAG-degrading lyases and subsequent electrophoretic profiling revealed the presence of three GAG species, i.e., hyaluronic acid (HA), heparan sulfate (HS) and galactosaminoglycans (GalAGs). A notable reduction of the total sulfated GAGs and a redistribution of the aortic GAG pattern were recorded in the WB as compared to the C group: a 25% and 10% increase in HA and HS, respectively, and an 11% decrease in GalAGs. Fine biochemical analysis of GalAGs at the level of constituent disaccharides with high-performance capillary electrophoresis revealed a notable increase of nonsulfated (18.0% vs. 10.7%) and a decrease of disulfated disaccharides (2.2% vs. 5.3%) in the WB aorta. This is the first study to report the redistribution of GAGs at the level of composition and their fine structural characteristics with implications for the endothelial dysfunction of the SHR.

Attenuation of alpha-adrenergic-induced vasoconstriction by dietary wild blueberries (Vaccinium angustifolium) is mediated by the NO-cGMP pathway in spontaneously hypertensive rats (SHRs)

Abstract The role of wild blueberries (WB) on key signaling steps of nitric oxide (NO) and cyclooxygenase (COX) pathways was examined in spontaneously hypertensive rats (SHRs) after eight weeks on a control (C) or an 8% w/w WB diet. Aortic rings from SHRs were stimulated with phenylephrine (Phe) in the absence or presence of inhibitors of: soluble guanylyl cyclase (sGC), phosphodiesterase-5 (PDE5), prostaglandin I2 (PGI2) synthase and thromboxane A2 (TXA2) synthase. Additionally, enzymatic activities in these pathways were determined by the concentration of NO, cyclic guanosine monophosphate (cGMP), PGI2 and TXA2. In the WB-fed SHR, attenuation of Phe-induced vasoconstriction was mediated by an increased synthesis or preservation of cGMP. Despite an increased release of PGI2 in the WB group, neither inhibition of PGI2 or TXA2 synthase resulted in a different response to Phe between the control and the WB rings. Hence, in the SHR, WB decrease Phe-mediated vasoconstriction under basal conditions by enhancing NO-cGMP signaling without a significant involvement of the COX pathway.