Objective To study the effect and mechanism of blueberry on regulating the mitochondrial inner membrane protein mitofilin/Mic60 in an in vitro model of metabolic dysfunction-associated liver disease (MAFLD). Methods L02 human hepatocytes were induced by free fatty acids (FFA) to establish MAFLD cell model. A normal group, a model group, an 80 μg/mL blueberry treatment group, a Mic60 short hairpin RNA (Mic60 shRNA) transfection group, and Mic60 knockdown combined with an 80 μg/mL blueberry treatment group were established. The intracellular lipid deposition was observed by oil red O staining, and the effect of different concentrations of blueberry pulp on the survival rate of L02 cells treated with FFA was measured by MTT assay. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglyceride (TG), total cholesterol (TC), superoxide dismutase (SOD) activity, glutathione (GSH) and malondialdehyde (MDA) contents were measured by visible spectrophotometry. The expression of reactive oxygen species (ROS) in hepatocytes was observed by fluorescence microscopy, and the mRNA and protein expression of Mic60 were detected by real-time quantitative PCR and Western blot analysis, respectively. Results After 24 hours of FFA stimulation, a large number of red lipid droplets in the cytoplasm of L02 cells was observed, and the survival rate of L02 cells treated with 80 μg/mL blueberry was higher. The results of ALT, AST, TG, TC, MDA and the fluorescence intensity of ROS in blueberry treated group were lower than those in model group, while the levels of SOD, GSH, Mic60 mRNA and protein in blueberry treated group were higher than those in model group. Conclusion Blueberry promotes the expression of Mic60, increases the levels of SOD and GSH in hepatocytes, and reduces the production of ROS, thus alleviating the injury of MAFLD hepatocytes and regulating the disorder of lipid metabolism.
Humans ; Blueberry Plants/chemistry* ; Hepatocytes/metabolism* ; Liver/metabolism* ; Liver Diseases/metabolism* ; Reactive Oxygen Species/metabolism* ; Superoxide Dismutase/metabolism* ; Superoxides/metabolism* ; Mitochondrial Membranes/metabolism* ; Mitochondrial Proteins/metabolism* ; Plant Extracts/pharmacology*

Blueberries are rich in phenolic compounds including anthocyanins which are closely related to biological health functions. The purpose of this study was to investigate the antioxidant activity of blueberry anthocyanins extracted from 'Brightwell' rabbiteye blueberries in mice. After one week of adaptation, C57BL/6J healthy male mice were divided into different groups that were administered with 100, 400, or 800 mg/kg blueberry anthocyanin extract (BAE), and sacrificed at different time points (0.1, 0.5, 1, 2, 4, 8, or 12 h). The plasma, eyeball, intestine, liver, and adipose tissues were collected to compare their antioxidant activity, including total antioxidant capacity (T-AOC), superoxide dismutase (SOD) activity and glutathione-peroxidase (GSH-PX/GPX) content, and the oxidative stress marker malondialdehyde (MDA) level. The results showed that blueberry anthocyanins had positive concentration-dependent antioxidant activity in vivo. The greater the concentration of BAE, the higher the T-AOC value, but the lower the MDA level. The enzyme activity of SOD, the content of GSH-PX, and messenger RNA (mRNA) levels of Cu,Zn-SOD, Mn-SOD, and GPX all confirmed that BAE played an antioxidant role after digestion in mice by improving their antioxidant defense. The in vivo antioxidant activity of BAE indicated that blueberry anthocyanins could be developed into functional foods or nutraceuticals with the aim of preventing or treating oxidative stress-related diseases.
Male ; Mice ; Animals ; Antioxidants/pharmacology* ; Blueberry Plants ; Anthocyanins/pharmacology* ; Mice, Inbred C57BL ; Superoxide Dismutase ; Plant Extracts/pharmacology* ; Superoxide Dismutase-1

OBJECTIVE: To study the protective effect of anthocyanins extracted from Vaccinium Uliginosum (VU) on retinal 661W cells against microwave radiation induced retinal injury. METHODS: 661W cells were divided into 6 groups, including control, model [661W cells radiated by microwave (30 mW/cm², 1 h)] and VU groups [661W cells pretreated with anthocyanins extracted from VU (25, 50, 100 and 200 µg/mL, respectively) for 48 h, and radiated by microwave 30 mW/cm², 1 h]. After treatment with different interventions, the cell apoptosis index (AI) was determined using Heochst staining; contents of malonaldehyde (MDA), glutataione (GSH), and activity of superoxide dismutase (SOD) were measured. mRNA expressions of nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase 1(HO-1) were detected by real time quantitative polymerase chain reaction, and the expression of HO-1 protein was examined by Western blot analysis. Nucleus and cytoplasm were separated and Nrf2 protein expression was further verified by Western blot analysis. RESULTS: There was significant difference in AI among the groups (F=322.83, P<;0.05). Compared with the control group, AI was significantly higher in the model group and was lower in 4 VU-pretreated groups (P<;0.05). Linear regression analysis showed the decline of AI was in a dose-dependent manner with VU treatment (r=0.8419, P<;0.05). The MDA and GSH contents of 661W cells in VU-treated groups were significantly lower than the model group (P<;0.05). Compared with the model group, the SOD activity in the VU-treated groups (50, 100 and 200 µg/mL) was significantly higher (all P<;0.05). The Nrf2 and HO-1 mRNA expressions were slightly increased after irradiation, and obviously increased in 100 µg/mL VU-treated group. After irradiation, the relative expressions of HO-1 and Nrf2 proteins in nucleus were slightly increased (P<;0.05), and the changes in cytoplasm were not obvious, whereas it was significantly increased in both nucleus and cytoplasm in the VU treatment groups. CONCLUSIONS Anthocyanins extracted from VU could reduce apoptosis, stabilize cell membrane, and alleviate oxidant injury of mouse retinal photoreceptor 661W cells. The mechanism might be through activating Nrf2/HO-1 signal pathway and inducing HO-1 transcription and translation.
Animals ; Anthocyanins/therapeutic use* ; Blueberry Plants/metabolism* ; Heme Oxygenase-1/metabolism* ; Mice ; Microwaves ; NF-E2-Related Factor 2/metabolism* ; Oxidative Stress ; RNA, Messenger/metabolism* ; Superoxide Dismutase/metabolism*

PURPOSE: Fruit and vegetable juices are known to be rich sources of antioxidants, which have beneficial effects on diseases caused by oxidative stress. The purpose of this study was to directly compare the antioxidant activities of fruit and vegetable juices marketed in Korea. METHODS: We analyzed four fruit juices, two vegetable juices, two yellow-green juices, and six mixed vegetable juices. Antioxidant activities were analyzed using 2,2-diphenyl-1-picrylhydrazyl (DPPH) test, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonate) (ABTS) test, and oxygen radical absorbance capacity (ORAC) assay. Protective effects against DNA damage were determined using an ex vivo comet assay with human lymphocytes. RESULTS: DPPH radical scavenging activities were in the following order: blueberry juice > mixed vegetable C juice > kale juice > mixed vegetable P juice > grape juice. ABTS radical scavenging activities were in the following order: blueberry juice > mixed vegetable C juice > grape juice > mixed vegetable P juice > kale juice. Peroxyl radical scavenging activities as assessed by ORAC assay were in the following order: blueberry juice > kale juice > mixed vegetable C juice > grape juice. Grape or blueberry juice showed strong abilities to prevent DNA damage in lymphocytes, and the difference between them was not significant according to the GSTM1/GSTT1 genotype. CONCLUSION: Antioxidant activities of fruit and vegetable juices and ex vivo DNA protective activity increased in the order of blueberry juice, grape juice, and kale juice, although the rankings were slightly different. Therefore, these juices rich in polyphenols and flavonoids deserve more attention for their high antioxidant capacity.
Antioxidants ; Blueberry Plant ; Brassica ; Comet Assay ; DNA Damage* ; DNA* ; Flavonoids ; Fruit and Vegetable Juices* ; Fruit* ; Genotype ; Humans ; Korea* ; Lymphocytes* ; Oxidative Stress ; Oxygen ; Polyphenols ; Vegetables ; Vitis

BACKGROUND: Chemopreventive effects and the underlying mechanisms of blueberry (Vaccinium spp.) are not clearly understood in human. We hypothesized blueberry would work via antioxidative and epigenetic modulation, which is similar to vitamin C. METHODS: We performed a pilot and non-inferiority study in healthy young women (n = 12), who consumed vitamin C (1 g/d) or 240 mL of blueberry juice (total polyphenols 300 mg and proanthocyanidin 76 mg/d) for 2 weeks. We analyzed 8-hydroxydeoxyguanosine (8-OHdG) and malondialdehyde (MDA) levels in their urine, and global and specific DNA methylation at the NAD(P)H quinone oxidoreductase 1 (NQO1), methylenetetrahydrofolate reductase (MTHFR), or DNA methyltransferase 1 (DNMT1) genes in their blood. RESULTS: Urinary 8-OHdG levels were reduced by blueberry consumption rather than by vitamin C. The methylation (%) of the MTHFR was significantly decreased in blueberry-consumers and the antioxidant-susceptible subgroup, whose urinary MDA levels were decreased by the intervention. We also found a positive correlation between changes of urinary 8-OHdG and of DNA methylation at the MTHFR or the DNMT1 (P < 0.05). However, the genetic polymorphism of the MTHFR (C677T in exon 4) did not affect any above markers. CONCLUSIONS: Blueberry juice shows similar anti-oxidative or anti-premutagenic activity to vitamin C and the potential as a methylation inhibitor for the MTHFR and the DNMT1 in human.
Ascorbic Acid* ; Blueberry Plant* ; DNA ; DNA Methylation ; Epigenomics* ; Exons ; Female ; Humans* ; Malondialdehyde ; Methylation ; Methylenetetrahydrofolate Reductase (NADPH2) ; Oxidative Stress ; Polymorphism, Genetic ; Polyphenols ; Vitamins*

BACKGROUND/OBJECTIVES: Stress-induced cognitive impairment is related to the suppression of hippocampal neurogenesis that results from an increase of oxidative stress. Therefore, the aim of this study was to investigate the effects of administration of a blueberry drink, having a high antioxidant power, on the cognitive performance of adult rats exposed to chronic mild stress. MATERIALS/METHODS: Twelve-week-old male Sprague-Dawley rats (n = 48) were randomly divided into four groups: control (CO), stress (ST), control + 5% blueberry drink (CO + B), and stress + 5% blueberry drink (ST + B). After eight weeks, the cognitive performance was assessed using a multiple T-maze water test. Levels of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and ascorbic acid were measured in the brain, and catecholamine concentrations were measured in plasma. RESULTS: The brain weights of the rats from the ST and ST + B groups were significantly lower than those of the rats from the CO and CO + B groups. The cognitive performance of the ST group was impaired when compared to that of the CO group. This impairment was significantly improved by the blueberry drink supplementation (P < 0.05). The brain SOD and CAT concentrations were not influenced by the stress or by the blueberry drink. However, the brain levels of GPx and ascorbic acid were significantly lower in the ST group than those in the CO group and were increased by the blueberry drink supplementation. The plasma catecholamine concentrations were affected by chronic mild stress and by the blueberry drink. The plasma norepinephrine and dopamine concentrations were decreased by the chronic stress and improved by the blueberry drink supplementation. The plasma epinephrine level was only influenced by the stress. CONCLUSION: These findings suggest that the blueberry drink may protect against the cognitive impairment induced by chronic mild stress.
Adult* ; Animals ; Ascorbic Acid ; Blueberry Plant* ; Brain ; Catalase ; Cats ; Cognition Disorders* ; Dopamine ; Epinephrine ; Glutathione Peroxidase ; Humans ; Male ; Neurogenesis ; Norepinephrine ; Oxidative Stress ; Plasma ; Rats* ; Rats, Sprague-Dawley ; Superoxide Dismutase ; Water ; Weights and Measures

Cryptosporidium and Cyclospora are well-known coccidian protozoa that can cause waterborne and foodborne diarrheal illnesses. There have been a few reports regarding contamination in different vegetables with Cryptosporidium, but no data are available regarding the sources of Cyclospora infections in Korea. In the present study, we collected 6 kinds of vegetables (perilla leaves, winter-grown cabbages, chives, sprouts, blueberries, and cherry tomatoes) from July 2014 to June 2015, and investigated contamination by these 2 protozoa using multiplex quantitative real-time PCR. Among 404 vegetables, Cryptosporidium and Cyclospora were detected in 31 (7.7%) and 5 (1.2%) samples, respectively. In addition, Cryptosporidium was isolated from all 6 kinds of vegetables, whereas Cyclospora was detected in 4 kinds of vegetables (except perilla leaves and chives). Cryptosporidium (17.8%) and Cyclospora (2.9%) had the highest detection rates in chives and winter-grown cabbages, respectively. Cryptosporidium was detected all year long; however, Cyclospora was detected only from October to January. In 2 samples (sprout and blueberry), both Cryptosporidium and Cyclospora were detected. Further investigations using TaqI restriction enzyme fragmentation and nested PCR confirmed Cryptosporidium parvum and Cyclospora cayetanensis, respectively. In conclusion, we detected C. cayetanensis in vegetables for the first time in Korea. This suggests that screening should be employed to prevent these protozoal infections in Korea.
Blueberry Plant ; Brassica ; Chive ; Cryptosporidium parvum ; Cryptosporidium* ; Cyclospora* ; Korea* ; Mass Screening ; Perilla ; Polymerase Chain Reaction ; Real-Time Polymerase Chain Reaction ; Vegetables*

PURPOSE: The objective of the present study was to determine whether fermentation can increase the protective effects of blueberry liquid in a high-fat diet-induced obese mice model. METHODS: Male C57BL/6J mice were fed a high-fat diet (HD, 60% fat, w/w,), HD supplemented with 10 ml/kg BW/day of blueberry liquid (BHD, blueberry high-fat diet), or HD supplemented with 10 ml/kg BW/day of fermented blueberry liquid (FBHD, fermented blueberry high-fat diet) for 10 weeks. RESULTS: There were significant decreases in the body, epididymal adipose tissue, and liver weights of blueberry-fed groups compared to HD, whereas there were no significant differences in food intake among the groups. Furthermore, blueberry liquid groups, especially fermented blueberry liquid, significantly attenuated the contents of hepatic triglycerides and total cholesterol induced by HD. Serum LDL-cholesterol was significantly lower in the BHD and FBHD-fed groups, whereas FBHD significantly increased the serum HDL-cholesterol level compared to the control. Concentrations of aspartate transaminase, alanine transaminase, and leptins in serum were also reduced by blueberry liquid supplementation. The mRNA expression of hepatic acetyl CoA carboxylase was significantly reduced in both the BHD and FBHD groups compared to HD. Furthermore, FBHD altered the mRNA expression level of hepatic lipolysis genes. CONCLUSION: In conclusion, these results suggest that blueberry, especially fermented blueberry liquid, may improve obesity-related abnormalities.
Acetyl-CoA Carboxylase ; Adipose Tissue ; Alanine Transaminase ; Animals ; Aspartate Aminotransferases ; Blueberry Plant* ; Cholesterol ; Diet, High-Fat ; Eating ; Fermentation ; Humans ; Leptin ; Lipolysis ; Liver ; Male ; Mice* ; Mice, Obese ; RNA, Messenger ; Triglycerides ; Weights and Measures

BACKGROUND/OBJECTIVES: Evidence indicates that berry anthocyanins are anti-atherogenic, antioxidant, and anti-inflammatory. However, berries differ vastly in their anthocyanin composition and thus potentially in their biological and metabolic effects. The present study compared hypolipidemic, antioxidant, and anti-inflammatory properties of blueberry (BB), blackberry (BK), and blackcurrant (BC) in a diet-induced obesity (DIO) mouse model. MATERIALS/METHODS: Male C57BL/6J mice were fed a high fat (HF; 35% fat, w/w) control diet or a HF diet supplemented with freeze-dried 5% BB, 6.3% BK or 5.7% BC for 12 weeks (10 mice/group) to achieve the same total anthocyanin content in each diet. Plasma lipids, antioxidant status and pro-inflammatory cytokines were measured. The expression of genes involved in antioxidant defense, inflammation, and lipid metabolism was determined in the liver, epididymal adipose tissue, proximal intestine, and skeletal muscle. Histological analysis was performed to identify crown-like structure (CLS) in epididymal fat pads to determine macrophage infiltration. RESULTS: No differences were noted between the control and any berry-fed groups in plasma levels of liver enzymes, insulin, glucose, ferric reducing antioxidant power, superoxide dismutase, and tumor necrosis factor α. However, BK significantly lowered plasma triglyceride compared with the HF control and other berries, whereas BC significantly reduced F4/80 mRNA and the number of CLS in the epididymal fat pad, indicative of less macrophage infiltration. CONCLUSIONS: The present study provides evidence that BB, BK and BC with varying anthocyanin composition differentially affect plasma lipids and adipose macrophage infiltration in DIO mice, but with no differences in their antioxidant capacity and anti-inflammatory potential.
Adipose Tissue ; Animals ; Anthocyanins ; Blueberry Plant* ; Cytokines ; Diet ; Fruit ; Glucose ; Humans ; Inflammation ; Insulin ; Intestines ; Lipid Metabolism ; Liver ; Macrophages ; Male ; Mice* ; Muscle, Skeletal ; Obesity* ; Plasma* ; RNA, Messenger ; Rubus* ; Superoxide Dismutase ; Triglycerides ; Tumor Necrosis Factor-alpha

OBJECTIVETo investigate whether the antioxidation and the regulation on the Extracellular Regulated Protein Kinases (ERK) signaling pathway are involved in the protective effects of blueberry on central nervous system.

METHODS30 Senescence-accelerated mice prone 8 (SAMP8) mice were divided into three groups and treated with normal diet, blueberry extracts (200 mg/kg•bw/day) and cyaniding-3-O-galactoside (Cy-3-GAL) (50 mg/kg•bw/day) from blueberry for 8 weeks. 10 SAMR1 mice were set as control group. The capacity of spatial memory was assessed by Passive avoidance task and Morris water maze. Histological analyses on hippocampus were completed. Malondialdehyde (MDA) levels, Superoxide Dismutase (SOD) activity and the expression of ERK were detected.

RESULTSBoth Cy-3-GAL and blueberry extracts were shown effective functions to relieve cellular injury, improve hippocampal neurons survival and inhibit the pyramidal cell layer damage. Cy-3-GAL and blueberry extracts also increased SOD activity and reduced MDA content in brain tissues and plasma, and increased hippocampal phosphorylated ERK (p-ERK) expression in SAMP8 mice. Further more, the passive avoidance task test showed that both the latency time and the number of errors were improved by Cy-3-GAL treatment, and the Morris Water Maze test showed significant decreases of latency were detected by Cy-3-GAL and blueberry extracts treatment on day 4.

CONCLUSIONBlueberry extracts may reverse the declines of cognitive and behavioral function in the ageing process through several pathways, including enhancing the capacity of antioxidation, altering stress signaling. Cy-3-GAL may be an important active ingredient for these biological effects.

Aging ; drug effects ; Animals ; Anthocyanins ; pharmacology ; Avoidance Learning ; Blueberry Plants ; chemistry ; Dietary Supplements ; Galactosides ; pharmacology ; Hippocampus ; drug effects ; metabolism ; Malondialdehyde ; metabolism ; Maze Learning ; Memory ; drug effects ; Mice ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Phosphorylation ; Plant Extracts ; pharmacology ; Superoxide Dismutase ; metabolism