Blueberry Treatment Research Articles

Inulin and Freeze-Dried Blueberry Intervention Lead to Changes in the Microbiota and Metabolites within In Vitro Studies and in Cognitive Function within a Small Pilot Trial on Healthy Children.

The relationship between the gut microbiota and cognitive health is complex and bidirectional, being significantly impacted by our diet. Evidence indicates that polyphenols and inulin can impact cognitive function via various mechanisms, one of which is the gut microbiota. In this study, effects of a wild blueberry treatment (WBB) and enriched chicory inulin powder were investigated both in vitro and in vivo. Gut microbiota composition and metabolites, including neurotransmitters, were assessed upon faecal microbial fermentation of WBB and inulin in a gut model system. Secondly, microbiota changes and cognitive function were assessed in children within a small pilot (n = 13) trial comparing WBB, inulin, and a maltodextrin placebo, via a series of tests measuring executive function and memory function, with faecal sampling at baseline, 4 weeks post-intervention and after a 4 week washout period. Both WBB and inulin led to microbial changes and increases in levels of short chain fatty acids in vitro. In vivo significant improvements in executive function and memory were observed following inulin and WBB consumption as compared to placebo. Cognitive benefits were accompanied by significant increases in Faecalibacterium prausnitzii in the inulin group, while in the WBB group, Bacteroidetes significantly increased and Firmicutes significantly decreased (p < 0.05). As such, WBB and inulin both impact the microbiota and may impact cognitive function via different gut-related or other mechanisms. This study highlights the important influence of diet on cognitive function that could, in part, be mediated by the gut microbiota.

The effects of curcumin and blueberry on axonal regeneration after peripheral nerve injury

The purpose of this study was to analyze the axonal regeneration and therapeutic effects of curcumin and blueberry administration following peripheral nerve injury using stereological, electron microscopic and electrophysiological methods. Animals in were assigned into one of four groups - control (Cont), injury (Inj), injury+curcumin (Cur) and injury+blueberry (Blue). Following the induction of sciatic nerve crush injury (75 Newtons for 5 s) in the Inj, Cur, and Blue groups, the rats in the Cur group received intraperitoneal injection of 30 mg/kg curcumin (Sigma C1386) and the rats in the Blue group received 4 g/kg blueberry by gavage over a four-week period. The rats in the Cont and Inj groups were not exposed to any substance. All animals were given standard chow. Sciatic functional index analyses were performed on the 14th and 28th days after injury, and electromyography (EMG) results were recorded. Stereological analysis of the nerve was performed under light microscopy. Light and electron microscopies were used for the histopathological evaluation of the sciatic nerve. Analysis of myelinated axon numbers revealed no significant differences between the Inj group and the Cur and Blue groups. However, a significant difference was observed between the Blue and Inj groups in terms of axonal areas. EMG test results differed between the Blue and the Inj groups (p < 0.05), but no significant difference was observed between the Inj and Cur groups. Electron microscopic analysis revealed protective effects of curcumin and blueberry treatment after injury. The use of the curcumin and blueberry may represent a supportive approach to the protection of nerve fibers after peripheral nerve crush injury.

Dose and Time Dependent Effect of Dietary Blueberries on Diabetic Vasculature is Associated with Gut Microbes

BackgroundEvidence from our lab and others showed the vascular beneficial effects of dietary blueberries. However, dose‐ and time‐dependent effect of dietary blueberries on vascular complications in diabetes and the role of gut microbes in mediating this effect are unknown. We hypothesize that gut microbes are associated with the dose‐ and time‐dependent vascular beneficial effects of dietary blueberries.Methods7‐week‐old db/db male mice (Jackson Lab), a widely‐used type 2 diabetic animal model, were fed a diet with or without freeze‐dried wild blueberry powder (FD‐BB) for 4 or 8 or 12 weeks (D4, D8, and D12). Diets contained 0%, 1.23% (B1), 2.46% (B2) and 3.7% (B3) of FD‐BB, which is equivalent to 0, ½, 1, and 2 human servings of wild blueberries. Age‐matched, non‐diabetic db/+ mice fed a standard diet for 4 or 8 or 12 weeks (C4, C8, and C12) were used as controls. After the treatment period blood glucose, body composition, glucose tolerance, and insulin tolerance were assessed. Vascular inflammation was determined by the binding of fluorescent labelled mouse monocytic WEHI78/24 cells to vascular endothelium and mRNA expression of vascular inflammatory chemokines and adhesion molecules (IL8/KC, MCP1/JE, ICAM1, VCAM1 and E‐selectin). Microbial profiling of cecum was done by 16s rRNA amplification.ResultsDiabetic mice (D4, D8 and D12) had increased blood glucose, impaired glucose and insulin tolerance, increased body fat, decreased lean body mass, enhanced binding of monocytes to the aortic vessel, and increased expression of selected inflammatory molecules as compared to their respective control mice (C4, C8 and C12). Blueberry treatment at all levels didn’t improve the metabolic parameters in diabetic mice. However, 3.7% FD‐BB treatment reduced the binding of WEHI78/24 monocyte to vascular endothelium and suppressed the mRNA expression of selected inflammatory molecules in diabetic mice (D4B3 vs D4, D8B3 vs D8, and D12B3 vs D12). Microbial profiling showed an alteration in the relative abundance of gut microbes at different taxonomic levels in diabetic mice as compared to their respective control mice. At the genera level, 7, 9 and 10 genera were altered after 4‐, 8‐ and 12‐weeks treatment in diabetic vs control mice. Blueberry supplementation at different dosages improved the relative abundance of selected gut microbes in diabetic mice (&gt;4 genera) at different time points. Specifically, blueberry treatment increased the abundance of commensal microbe Bifidobacterium and decreased the abundance of opportunistic Clostridium genera in diabetic mice.ConclusionThe beneficial effects of dietary blueberries on diabetic vasculature and gut microbiome varies with the doses and duration of treatment. In the present study, a dose of 3.7% FD‐BB in diet (equivalent to 1.5 human servings) was shown to exert potential beneficial effects on diabetic vasculature and is associated with specific gut microbes.

Blueberry treatment administered before and/or after lipopolysaccharide stimulation attenuates inflammation and oxidative stress in rat microglial cells

ABSTRACT Microglia are key regulators of inflammation and oxidative stress (OS) in the CNS. Microglia activation can lead to chronic inflammation, OS, and neurodegeneration. Blueberries (BB) reduce inflammation and OS when administered to microglia before stressors such as lipopolysaccharide (LPS), but the therapeutic value of BBs administered after activation by stressors has not been examined. Therefore, this study investigated the differential effects of pre-, post-, and pre-/post-BB on inflammation and OS in LPS-activated microglia. Rat microglia were pretreated with BB (0.5 mg/mL) or control media (C) for 24 hours, incubated overnight with LPS (0 or 200 ng/mL), and post-treated with BB or C for 24 hours. Biomarkers of inflammation (e.g. nitrite [NO2 −], tumor necrosis factor-ɑ [TNFɑ], inducible nitric oxide synthase [iNOS], cyclooxygenase-2 [COX-2], phosphorylated IκB-α [pIκB-ɑ]) and OS (e.g. NADPH oxidase [NOX2]) were assessed. LPS increased NO2 −, TNFɑ, COX-2, iNOS, pIκB-ɑ, and NOX2 compared to non-stressed conditions (P < 0.05), however BB before and/or after LPS significantly reduced these markers compared to no BB (P < 0.05). Pre-BB was more effective than post-BB at reducing LPS-induced NO2 −, TNFɑ, and COX-2 (P < 0.05). Pre-BB was also more effective than pre-/post-BB at attenuating LPS-induced NO2 − and TNFɑ (P < 0.05). All BB treatments were equally effective in reducing LPS-induced iNOS, pIκB-ɑ, and NOX2. Results suggest that BBs can target the downstream events of LPS-induced microglial activation and prevent stressor-induced neuroinflammation and OS. Furthermore, BBs may not need to be present prior to microglial activation for beneficial effects, suggesting that dietary interventions may be effective even after initiation of disease processes. Graphical . Cascade of inflammatory and OS-inducing events associated with self-propelling microglial activation by LPS and the effects of blueberry (0.5 mg/mL) administered before and/or after LPS on these processes (blue arrows). BB, blueberry; COX2, cyclooxygenase-2; IκB-ɑ, inhibitor kappa-B-ɑ; iNOS, inducible nitric oxide synthase; LPS, lipopolysaccharide; NF-κB, nuclear factor kappa-B; NO, nitric oxide; NOX2, NADPH oxidase; OS, oxidative stress; ROS, reactive oxygen species; TNFɑ, tumor necrosis factor-ɑ

Blueberry extract improves redox balance and functional parameters in the right ventricle from rats with pulmonary arterial hypertension.

Pulmonary arterial hypertension (PAH) is a disease characterized by increased pulmonary vascular resistance and right ventricle (RV) failure. In this context, oxidative stress is an essential element contributing to PAH's pathophysiology. Thus, blueberry (BB), which has a high antioxidant capacity, emerges as a natural therapeutic approach in PAH. This work evaluated the effect of BB extract on redox balance in RV in a PAH's animal model. Male Wistar rats (200 ± 20g) (n = 72) were randomized into eight groups: control (CTR); monocrotaline (MCT); CTR and MCT treated at doses of 50, 100, and 200mg/kg BB. PAH was induced by administration of MCT (60mg/kg, intraperitoneal). Rats were treated with BB orally for 5weeks (2weeks before monocrotaline and 3weeks after monocrotaline injection). On day 35, rats were submitted to echocardiography and catheterization, then euthanasia and RV harvesting for biochemical analyses. RV hypertrophy, observed in the MCT groups, was reduced with BB treatment. MCT elevated RV systolic pressure and pressure/time derivatives, while the intervention with BB decreased these parameters. PAH decreased RV output and pulmonary artery outflow acceleration/ejection time ratio, while increased RV diameters, parameters restored by BB treatment. Animals from the MCT group showed elevated lipid peroxidation and NADPH oxidase activity, outcomes attenuated in animals treated with BB, which also led to increased catalase activity. Treatment with BB partially mitigated PAH, which could be associated with improvement of RV redox state. Such findings constitute an advance in the investigation of the role of BB extract in chronic progressive cardiovascular diseases that involve the redox balance, such as PAH.

Blueberry Treatment Administered Before and/or After Lipopolysaccharide Attenuates Neuroinflammation and Oxidative Stress in Rat Microglial Cells

ObjectivesNeuroinflammation and oxidative stress (OS) are implicated as mediators of cognitive decline and neurodegenerative diseases. Microglia, the primary immune cells of the central nervous system (CNS), are key regulators of inflammation and OS in the CNS. Blueberries (BB) are rich in anti-inflammatory and antioxidant polyphenols and reduce inflammation and OS when administered to microglia before pathogenic stimuli such as lipopolysaccharide (LPS). However, the therapeutic value of BBs administered to microglial cells after activation by LPS on inflammation and OS has not been examined. This study investigated the potential differential effects of pre-, post-, and pre-/post-BB on LPS-induced inflammation and OS in rat microglia. MethodsRat microglia were pre-treated with freeze-dried BB diluted in media (0.5 mg/mL) or control media (C) for 24 h, incubated overnight with LPS (0 or 100 ng/mL), and post-treated with BB or C for 24 h. Standard immunochemical techniques were used to measure biomarkers of inflammation and OS, including nitrite, tumor necrosis factor alpha (TNFɑ), inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2). Data were analyzed using two-way ANOVAs with treatment group (pre-/post-BB) and LPS exposure as experimental factors. Post-hoc testing was performed using Tukey’s test to determine differences among groups. ResultsPreliminary results showed that all BB treatments (pre-, post- and pre-/post-BB) reduced LPS-induced nitrite and TNFɑ relative to no BB (P < 0.01). Pre-BB was more effective at reducing nitrite than post-BB (P < 0.01), although neither was significantly different than pre-/post-BB. In contrast, the attenuating effects of pre-BB and post-BB on TNFɑ were not different from each other, although both treatments were more beneficial than pre-/post-BB (P < 0.001). Compared to no BB, both pre-BB and post-BB, although not pre-/post-BB reduced LPS-induced iNOS expression (P < 0.05), and only pre-BB attenuated COX2 expression (P < 0.01). ConclusionsResults suggest that BBs can target the downstream effects of microglial activation in addition to preventing stressor-induced neuroinflammation and OS. Therefore, although potentially more effective, BBs may not need to be present prior to microglial activation for beneficial effects. Funding SourcesSupported by USDA intramural funding.

Blueberry extract as a potential pharmacological tool for preventing depressive-like behavior and neurochemical dysfunctions in mice exposed to lipopolysaccharide

ABSTRACT Objective: Major depressive disorder is a debilitating and recurrent psychiatric disorder. Blueberries have several biological properties, including neuroprotective effects, through antioxidant and anti-inflammatory actions. The aim of this study was to evaluate the effect of blueberry extract on depressive-like behavior and lipopolysaccharide (LPS)-induced neurochemical changes. Methods: Mice were pretreated with vehicle, fluoxetine (20 mg/kg) or blueberry extract (100 or 200 mg/kg) intragastrically for seven days before intraperitoneal LPS (0.83 mg/kg) injection. Twenty-four hours after LPS administration, mice were submitted to behavioral tests. Oxidative stress and neuroinflammatory parameters were evaluated in the cerebral cortex, hippocampus, and striatum. Results: Our data showed that blueberry extract or fluoxetine treatment protected against LPS-induced depressive-like behavior in tail suspension and splash tests (P < 0.05), without changes in locomotor activity (P > 0.05). LPS induced an increase in the levels of reactive oxygen species (P < 0.001), nitrite (P < 0.05) and thiobarbituric acid reactive substances (P < 0.01), as well as a reduction in total sulfhydryl content (P < 0.05) and catalase activity (P < 0.05) in brain structures; blueberry extract restored these alterations (P < 0.05). In addition, blueberry extract attenuated the increase in tumor necrosis factor-alpha (TNF-α) levels induced by LPS administration (P < 0.05). Conclusion: This study showed that blueberry extract exerted antidepressant-like effects, protected the brain against oxidative damage, and modulated TNF-α levels induced by LPS.

Beneficial Effects of Blueberry Extracts on Adult Human Neural Progenitor Cells

Abstract Objectives Aging-induced declines in neurogenesis and stem cell functions play a key role in the pathophysiology of age-associated neuronal disorders, and oxidative stress and inflammation are the major contributing factors. Blueberries, rich in polyphenols, have been shown to decrease oxidative stress, inflammation, and improve cognitive function in aged animals and humans. The current study investigated the effects of blueberry (BB) treatments on the viability, proliferation, fate choice, and expression of oxidative stress and inflammation markers of adult human neural progenitor cells (AHNPs), to establish mechanisms underlying the beneficial effects. Methods AHNPs derived from control and Parkinson's disease (PD) patients were pre-treated with various concentrations of freeze-dried BB extract for 7–10 days. Their viability and proliferation were examined using Trypan Blue exclusion and ethynyl-deoxyuridine (EdU) assay, respectively. Some treated cells were labeled with neuronal markers to examine their differentiation. Expression of oxidative stress and inflammation markers such as iNOS and Nox-2 was evaluated using western blot analysis. Whether BB could protect AHNPs from stress induced by dopamine (DA) was investigated by exposing cells to DA for 2–4 hours prior to evaluating their viability, proliferation, calcium buffering, and expression of oxidative stress and inflammation markers. Results Our data show that while BB treatment did not significantly affect the viability of control- and PD-AHNPs, BB extract significantly increased the proliferation rate of PD-AHNPs at higher concentrations. Additionally, BB treatment was able to revive the decrease in cell viability and proliferation following cellular stress induced by DA, indicating a potential neuroprotective effect. BB treatment was also able to protect against the harmful effects on Ca2+ buffering and reduce the increased expression levels of both iNOS and Nox-2 induced by DA exposure. Conclusions Blueberry extracts confer anti-oxidative, anti-inflammatory and neuroprotective effects on control and Parkinson's AHNPs, suggesting a role for dietary nutrients in helping to slow progression of neural dysfunction and neurodegenerative disease during aging. Funding Sources Supported by USDA/ARS intramural grant, and the Robert and Margaret Patricelli Family Foundation.

Dose Dependent Effects of Whole Blueberry on Cartilage Health and Pain in a Monosodium Iodoacetate (MIA) Induced Rat Model of Osteoarthritis

Abstract Objectives The purpose of this study was to examine the dose-dependent effects of whole blueberry in a monosodium iodoacetate (MIA) induced rat model of Osteoarthritis (OA). There is no known cure for OA and the present pharmacological treatment options are limited and associated with adverse side effects. We hypothesize that the addition of whole blueberry into the diets of rats with MIA induced OA will reduce pain behavior and improve cartilage health. Methods A total of forty, 45-day-old female CD rats were used for this study. Thirty rats were injected with MIA to induce joint destruction associated with OA. Ten rats served as control without induction of joint destruction. The MIA induced rats were randomized into three groups consisting of 10 animals. All groups were fed a casein-based diet with two of the MIA induced groups receiving an addition of whole blueberry powder at 5% and 10%, respectively. All groups were fed the respective diets for 46 days. The animals were weighted weekly throughout the study period and food intake monitored and recorded. Fasted blood specimens and other tissues of interest were collected after euthanasia for analysis. Mechanical allodynia was assessed at four time points throughout the study to evaluate changes in pain behaviors. Results There were no difference in body weight between the four groups at baseline. After 46 days of feeding there was no significant change in body weight in either of the blueberry fed groups in comparison to the MIA rats. Plasma hyaluronic acid levels were higher in the MIA group in comparison to the blueberry treated groups. We observed a dose dependent decrease in hyaluronic acid levels with blueberry treatment, respectively. At baseline, the MIA group exhibited significant (P ≤ 0.05) mechanical allodynia compared to the control group. Mechanical allodynia was significantly (P ≤ 0.05) reduced after 40 days in the 10% whole blueberry treated group. Conclusions Incorporation of whole blueberry into the diets of rats with MIA-induced OA reduced hyaluronic acid concentration and pain sensitivity. These results suggest that whole blueberry may be an effective therapeutic alternative for improving joint health and alleviating pain associated with OA. Funding Sources Research Enhancement Program, Texas Woman's University.

Effect of Whole Blueberry on Bone Biomarkers in Postmenopausal Women with Mild-to-Moderate Bone Loss (P01-029-19)

ObjectivesTo examine whether daily incorporation of 40 grams of whole blueberry into the diets of postmenopausal women with mild to moderate degree of bone loss will positively influence indices of bone metabolism. MethodsIn a randomized, double-blind control trial, 40 postmenopausal women with mild to moderate degree of bone loss, not on hormone or other bone therapies, were randomized to either consume 40 g of lyophilized blueberry powder (LBP) (n = 20) or a placebo powder (n = 20) daily for 90 days. Plasma bone biomarkers including sclerostin, bone-specific alkaline phosphatase (B-ALP), receptor activator of nuclear factor kappa-Β ligand (RANKL), tartrate-resistant acid phosphatase (TRAP5) and inflammatory marker, interleukin(IL)-6 was assessed at baseline, mid-point (45 days) and final (90 days). Bone density and body composition was assessed at baseline and final visits only. Fracture risk assessment tool (FRAX) was utilized to determine major osteoporotic fracture risk. ResultsA total of 32 participants completed the study with an attrition rate of 20%. A significant increase of B-ALP was observed at mid-point in the blueberry treatment group, indicating increased bone formation, however the increase was not maintained at final point. In addition, a more pronounced decline in RANKL was observed at final point in the LBP group as compared to the placebo group. A slight decline in the probability of a major osteoporotic fracture was observed in the LBP group, while an increase of such risks was observed in the placebo group. Moreover, in the placebo group, a significant increase in IL-6 was observed at final point, but the LBP group maintained their IL-6 throughout the study with a declining trend from baseline to final point. Besides bone metabolism parameters, body weight was decreased significantly in the blueberry group at final point, along with a more pronounced decrease in %body fat as compared to the placebo group. No changes in bone densities were observed, possibly due to the short study duration. ConclusionsThe findings of the study suggest that blueberry consumption may have nominal protective effects on bone in postmenopausal women who have mild to moderate bone loss. A longer study duration may more favorably influence bone metabolism and bone density in this population. Funding SourcesUS Highbush Blueberry Council and Research Enhancement Program at Texas Woman’s University.

Blueberry Consumption Improves Glycemic Control, Triglycerides and Liver Enzymes in US Veterans with Type 2 Diabetes (P06-124-19)

ObjectivesThe study investigated the effects of blueberry consumption on biomarkers of glycemic control in US veterans with type 2 diabetes. MethodsIn a double-blind, placebo-controlled parallel study, fifty-five men (mean baseline characteristics: 67 years; weight, 103 kg; body mass index, 34 kg/m2) with type 2 diabetes were randomly assigned to 1 of 2 treatment groups for 8 weeks. The treatment groups were either 22 g of a freeze-dried whole blueberry powder (equivalent to 1 cup of fresh blueberries; containing 845 mg phenolics and 470 mg anthocyanins) or 22 g of a blueberry placebo treatment (matched in energy and carbohydrate content to the blueberry treatment). The study participants were asked to consume 11 g of freeze-dried (equivalent to ½ cup of fresh blueberries) blueberry powder or placebo, reconstituted with 240 ml water, with their morning and evening meals along with their typical diet. Fasting blood samples were collected at the beginning and at the end of the study. ResultsFructosamine (275.5 ± 4.1 μmol/L vs. 292.4 ± 7.9 μmol/L, respectively; P = 0.04) and hemoglobin A1C (7.1 ± 0.1% vs. 7.5 ± 0.2%, respectively; P = 0.03) were significantly lower for study participants consuming blueberries for 8 weeks compared with the placebo. In addition, triglycerides (160.6 ± 13.1 mg/dl vs. 183.9 ± 10.7 mg/dl, respectively; P = 0.03), aspartate transaminase (AST) (23.2 ± 1.4 units/L vs. 30.5 ± 2.7 units/L, respectively; P = 0.02) and alanine transaminase (ALT) (35.6 ± 1.5 units/L vs. 48.3 ± 2.9 units/L, respectively; P = 0.0003), were significantly lower for those consuming blueberries compared with the placebo. Glucose, insulin, total cholesterol, LDL cholesterol, HDL cholesterol and body weight were not significantly different after 8 weeks of consumption of blueberries compared with the placebo. ConclusionsConsumption of 22 g of freeze-dried blueberries daily for 8 weeks results in better glycemic control, such as lowering of fructosamine and hemoglobin A1C. These results along with lower triglyceride concentrations show improvement in the liver enzymes, AST and ALT, which may beneficially affect the cardiometabolic health status of men with type 2 diabetes. Funding SourcesThe U.S. Highbush Blueberry Council; the study is the result of work supported with resources and the use of facilities at the Stratton VA Medical Center, Albany, NY, USA.

Anti-Inflammatory Action of Blueberry Polyphenols in HIG-82 Rabbit Synoviocytes.

Rheumatoid arthritis (RA) is an autoimmune and chronic inflammatory disease resulting in joint destruction and disability in the adult population. The etiology of RA is not well understood and presently there is no known cure for this disease. The accumulation and proliferation of fibroblast-like synoviocytes may be involved in cartilage destruction. Both in vitro and in vivo studies support an anti-inflammatory role of dietary polyphenols, the bioactive constituents found in fruits and vegetables. The objective of this study was to investigate the anti-inflammatory role of blueberry polyphenols (BBPs) using rabbit synoviocytes stimulated with tumor necrosis factor alpha (TNFα). Rabbit synoviocytes (HIG-82) were treated with varying doses of BBPs and stimulated with TNFα. Stimulation of rabbit synoviocytes with the proinflammatory cytokine TNFα increased cell proliferation by ∼19% compared with the nonstimulated control. Cell proliferation was significantly decreased in a dose-dependent manner by the treatment with BBPs. Post-TNFα stimulation, cells treated with BBPs resulted in decreases in interleukin 1 beta and nuclear factor kappa B (NFκB) concentration. Reverse transcription-polymerase chain reaction analysis showed that matrix metalloproteinase 3 increased fivefold in the control TNFα-stimulated group, but was decreased by threefold in the blueberry treatment group. These results suggest that downregulation of proinflammatory cytokines and transcription factor NFκB by naturally occurring bioactives such as BBPs may be a potential therapeutic strategy for reducing inflammation associated with RA.

Blueberries Improve Pain, Gait Performance, and Inflammation in Individuals with Symptomatic Knee Osteoarthritis

Osteoarthritis (OA) is the most common joint disorder in the world and is the most frequent cause of walking related disability among older adults in the US, which brings a significant economic burden and reduces quality of life. The initiation and development of OA typically involves degeneration or progressive loss of the structure and function of articular cartilage. Inflammation is one of the major drives of the progression of OA. Dietary polyphenols have been studied for their anti-inflammatory properties and potential anabolic effects on the cartilage cells. Blueberries are widely consumed and are high in dietary polyphenols, therefore regular consumption of blueberries may help improve OA. The purpose of the present study was to examine the effect of freeze dried whole blueberries on pain, gait performance, and inflammation in individuals with symptomatic knee OA. In a randomized, double-blind trial, adults age 45 to 79 with symptomatic knee OA, were randomized to either consume 40 g freeze-dried blueberry powder (n = 33) or placebo powder (n = 30) daily for four months. Blood draws and assessment of pain and gait were conducted at baseline, two months, and four months. Western Ontario McMaster Osteoarthritis Index (WOMAC) questionnaires were used to assess pain and GAITRite® electronic walkway was used to evaluate gait spatiotemporal parameters. WOMAC total score and sub-groups, including pain, stiffness, and difficulty to perform daily activities decreased significantly in the blueberry treatment group (p < 0.05), but improvement of WOMAC total score and difficulty to perform daily activities were not observed in the placebo group. Normal walking pace single support percentage for both limbs increased (p = or < 0.007), while double support percentage for both limbs decreased in the blueberry treatment group (p = or < 0.003). No significant changes were observed in plasma concentrations of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, IL-10, IL-13, matrix metalloproteinases (MMP)-3, MMP-13, and monocyte chemoattractant protein-1 (MCP-1) in both treatment groups. However, an increasing trend for IL-13 concentration and a decreasing trend in MCP-1 concentration were noted in the blueberry group. The findings of this study suggest that daily incorporation of whole blueberries may reduce pain, stiffness, and difficulty to perform daily activities, while improving gait performance, and would therefore improve quality of life in individuals with symptomatic knee OA.

The influence of juicing on the appearance of blueberry metabolites 2 h after consumption: a metabolite profiling approach.

The consumption of berries has been linked to decreased risk of degenerative disease. Berries are regularly processed into juices. It is largely unknown how the juicing process affects the bioavailability of metabolites. As metabolomics has shown to be a valuable nutritional tool to study global metabolite differences, the aim of this study was to investigate the effect of juicing on the relative appearance of blueberry metabolites in humans using metabolomics. Nine healthy subjects consumed 250 g of fresh blueberries either as the whole fruit or after juicing, and provided blood and urine samples before and 2 h after intake in a cross-over design. Samples underwent metabolite profiling using LCMS, and data were mined with multivariate analysis. Overall, <12 % of all ions detected were significantly influenced by blueberry treatment (P<0·05). Partial least-squared discriminant analysis models of post-treatment samples revealed good discrimination. In urinary samples, whole blueberry treatment resulted in 108 ions that were significantly higher compared with juiced treatment (positive and negative mode combined), whereas only eight were significantly higher after juiced treatment. Examples of putative annotations included metabolites of ferulic and caffeic acids, several phenolic metabolites conjugated to sulphate, glycoside or glucuronide and fatty acyl derivatives, which were of higher intensity after whole blueberry treatment. In conclusion, consumption of whole blueberries resulted in a higher range of phenolic and other metabolites in plasma and urine samples 2 h after consumption. Both whole and juiced blueberries resulted in very similar metabolite profiles at 2 h, although this was the only time point measured.

Cognitive response to fish oil, blueberry, and combined supplementation in older adults with subjective cognitive impairment

Given evidence that eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and anthocyanin-rich blueberries provide neurocognitive benefit, we investigated long-term supplementation in older adults with cognitive complaints. In a 24-week randomized, double-blind, placebo-controlled trial, elderly men and women received daily fish oil (FO) or blueberry (BB) or both. Diet records confirmed that participants reduced background consumption of EPA, DHA, and anthocyanins as prescribed. Erythrocyte EPA + DHA composition increased in the FO groups (p = 0.0001). Total urinary anthocyanins did not differ between the groups after supplementation but glycoside and native (food) forms increased only in the BB-supplemented groups. The FO (p = 0.03) and BB (p = 0.05) groups reported fewer cognitive symptoms, and the BB group showed improved memory discrimination (p = 0.04), indicating that supplementation improved cognition. Cognitive benefit in the BB group was associated with the presence of urinary anthocyanins reflecting recent BB intake but not with anthocyanin metabolites. However, combined FO + BB treatment was not associated with cognitive enhancement as expected.

Blueberry Phenolics Reduce Gastrointestinal Infection of Patients with Cerebral Venous Thrombosis by Improving Depressant-Induced Autoimmune Disorder via miR-155-Mediated Brain-Derived Neurotrophic Factor.

Cerebral venous thrombosis (CVT) often causes human depression, whereas depression-induced low immunity makes the patients susceptible to gastrointestinal infection. Blueberry possesses antidepressant properties which may improve autoimmunity and reduce gastrointestinal infection. Brain-derived neurotrophic factor (BDNF) performs antidepressant function and can be regulated by miR-155, which may be affected by blueberry. To explore the possible molecular mechanism, blueberry compounds were analyzed by high-performance liquid chromatography. Activity of compounds was tested by using HT22 cells. The present study tested 124 patients with CVT-induced mild-to-moderate depressive symptoms (Center for Epidemiologic Studies—Depression Scale [CES-D] ≥16) and gastrointestinal infection. Patients were randomly assigned to blueberry extract group (BG, received 10 mg blueberry extract daily) and placebo group (PG, received 10 mg placebo daily). After 3 months, depression, gastrointestinal infection and lipid profiles were investigated. Serum miR-155 and BDNF were measured using real-time quantitative polymerase chain reaction and or Western Blot. Blueberry treatment improved depressive symptoms and lipid profiles, and also reduced gastrointestinal infection in the BG group (P < 0.05) but those of the PG group (P = 1). These changes were paralleled by increase in serum levels of BDNF and miR-155 (P < 0.05). HPLC analysis showed that blueberry extracts were the main phenolic acids with 0.18, 0.85, 0.26, 0.72, 0.66, 0.4,1, and 1.92 mg/g of gentisic acid, chlorogenic acid, [2]-epicatechin, p-coumaric acid, benzoic acid, p-anisic acid, and quercetin in blueberry extracts, respectively. Phenolics in blueberry are possible causal agents in improving antidepressant activity and reducing gastrointestinal infection. Administration of blueberry increased BDNF expression and miR-155. Blueberry cannot affect BDNF level when miR-155 is overexpressed or inhibited. Phenolics from blueberry reduced gastrointestinal infection of patients with CVT by improving antidepressant activity via upregulation of miR-155-mediated BDNF.

The effects of blueberry anthocyanins on histone acetylation in rat liver fibrosis.

To determine the effects ofanthocyanins from blueberries on hepatic stellate cell (HSCs-T6) and on histone acetylation during liver fibrosis induced by CCl4 in rats. Fifty male SD rats weighing 180 ± 20g were randomly placed into a control group, a hepatic fibrosis group, a blueberry treatment group, a blueberry intervention group, and a natural recovery group. After the rats were sacrificed, the livers and the liver indexes were measured, and the pathological changes were observed by HE staining and Masson staining. The blood was analyzed for the four indexes of liver fibrosis and liver function; nucleoprotein from liver tissues and karyoplasm were isolated to determine the expression of acH3K9, acH3K14, and acH3K18 by Western blotting.Compared with the lethal rate of the control group, the median lethal rate of HSCs-T6 cells treated with a the 50μmol/L concentration was 66.94% (P < 0.05). The protein expression on α-SMA, type I collagen, TIMP1 significantly decreased (P < 0.05) following treatment with 50 ug/ml of anthocyanin for 36 h; moreover, the expression of acH3K9, acH3K14 and acH3K18 modification were up-regulated (P < 0.05). Furthermore, compared with the liver in the model group, the liver in the intervention group showed the most obvious improvement (P < 0.01), and its karyoplasm had increased expression of acH3K9, acH3K14 and acH3K18 (P<0.01).Regulating histone acetylation could improve liver function and liver fibrosis indexes in rats with hepatic fibrosis. The mechanism might be related to certain genes that promote apoptosis, so as to inhibit the effect of anti hepatic fibrosis.

Blueberries inhibit cyclooxygenase-1 and cyclooxygenase-2 activity in human epithelial ovarian cancer.

Ovarian cancer (OC) is the sixth and eighth leading cause of cancer mortality among women in developed and developing countries, respectively. Medical therapy is the main method for the treatment of OC. However, drug toxicity and the marked side effects of chemotherapy limit the usage and therapeutic results of the treatments. Therefore, the identification of multi-target agents with few side effects and high effectiveness is required. Traditional Chinese medicine has been used clinically to treat various types of cancer for thousands of years and is considered to possess multiple components and agents, which exert efficient therapeutic functions with few side effects. Although blueberries have previously been used to treat various types of cancer, the effect on OC and precise molecular mechanism of function of the fruit remains unknown. Cyclooxygenase (COX)-1 and COX-2 have been reported to be the biomarkers of OC. Blueberries may affect the progression of OC by affecting COX levels. To investigate the issue, COX-1 and COX-2 were overexpressed or silenced in ovarian cancer SKOV3 cells. The effect of blueberries on SKOV3 cell viability was determined by an MTT assay. Furthermore, a mouse model for OC was established. The results indicated that blueberries inhibited the proliferation of OC cells by downregulating the levels of COX-1 and COX-2. Blueberry (400 mg daily) consumption reduced tumor size significantly in mice with OC compared with the control without blueberry treatment (P<0.05). The results suggest that blueberries should be used to develop a potential non-pharmaceutical therapy for OC.

Investigation on the Role of BDNF intheBenefits of Blueberry Extracts for theImprovement of Learning and Memory inAlzheimer's Disease Mouse Model.

Blueberry (BB) can provide a wide range of antioxidant benefits for AD. There is evidence that BB extracts could improve brain functions. However, the details are still unknown. In the present study, we aimed to investigate the possible mechanism involved in the improvement of learning and memory capacity from BB extracts in AD. APP/PS1 transgenic mice were fed BB extracts for 16 weeks. The capacity of learning and memory was assessed by Morris water maze (MWM) test, and long-term potentiation (LTP) was determined to evaluate hippocampal neuronal plasticity at the end of administration. Pathological changes in the brain were observed, and the expressions of brain-derived neurotrophic factor (BDNF) and extracellular signal-related kinase (ERK1/2) were determined to explore the mechanism of BB extract-induced benefits. AD mice exhibited more difficulties to learn and remember the exact position of the platform in the MWM test. The data showed that AD mice lacked effective learning in the platform search. In contrast, AD mice exhibited better performance both in the training phase and probe test of MWM after the BB treatment. Moreover, LTP was enhanced and the neuron loss was alleviated with BB treatment, while we did not find any obvious effect on the elimination of amyloid-β. In the AD mice, the expression of ERK1/2 was significantly increased (p < 0.05), while the level of BDNF was decreased (p < 0.05). BB treatment was beneficial for the improvement of learning and memory of AD, and these effects might be related to the regulation of BDNF.

A serving of blueberry (V. corymbosum) acutely improves peripheral arterial dysfunction in young smokers and non-smokers: two randomized, controlled, crossover pilot studies.

Several studies have documented the important role of polyphenol-rich foods in the modulation of vascular remodelling and function. This study aimed to evaluate the capacity of a single portion of blueberry (V. corymbosum) to acutely improve peripheral arterial dysfunction in a group of young volunteers. Twenty-four healthy males (12 non-smokers and 12 smokers) were recruited for two different randomized, controlled, crossover pilot acute studies. In the first study, non-smokers were exposed to a control treatment (C; 300 mL of water with sugar) and a blueberry treatment (BB; 300 g of blueberry). In the second study, smokers underwent 3 different protocols: (1) - smoking treatment (S); (2) - control treatment (CS; 300 mL of water with sugar + smoking); (3) - blueberry treatment (BS; 300 g of blueberry + smoking). Each treatment (1 day long) was separated by a one week washout period. Blood pressure, peripheral arterial function (reactive hyperemia index, RHI, a marker of endothelial function) and arterial stiffness (digital augmentation index, dAix and dAix normalized by considering a heart rate of 75 bpm, dAix@75) were measured before and after each treatment. In the first study, the consumption of blueberry and control treatment acutely increased peripheral arterial function in the group of non-smokers. The improvement in RHI was higher and significantly different after blueberry treatment compared to the control treatment (54.8 ± 8.4% BB vs. 28.2 ± 8.3% C; p = 0.01). No effects were observed for markers of arterial stiffness, blood pressure and heart rate. Acute cigarette smoke significantly increased blood pressure and heart rate, while no significant effect was registered in peripheral arterial function and stiffness. The intake of blueberry and control treatment before a cigarette did not counteract the increase in blood pressure and heart rate, while it significantly improved peripheral arterial function. In particular, a significant increase was observed following BS (35.2 ± 7.5% RHI; p = 0.02) and CS treatments (34.6 ± 11.9% RHI; p = 0.02) when compared to only smoking treatment. No difference between BS and CS was detected. In conclusion, the intake of blueberry and control treatments acutely improved peripheral arterial dysfunction both in smoker and in non-smoker subjects. Further studies should be performed to confirm the results obtained and reveal the potential mechanisms of blueberry in the improvement of endothelial function.