OBJECTIVETo study the protective effect of Blueberry against rat hepatic fibrosis and the effect of Blueberry on HO-1 expression patterns.

METHODSA total of 45 SD rats were randomly divided into five groups namely control group (group A), model group (group B), blueberry group (group C), Dan-shao-hua-xian (DSHX) capsule group (group D) and blueberry +Dan-shao-hua-xian group (group E). Fibrous liver models in rats were induced by subcutaneous injection of CCl4 and high-lipid/low-protein diet for 8 weeks except the control group which accepted saline alone. The level of alanine aminotransferase (ALT) in serum was examined. The activities of superoxide dismutase (SOD) and malondialdehyde (MDA) in liver homogenates were determined. by the xanthine oxidase method and the thiobarbituric acid method. The pathology of hepatic fibrosis was evaluated by hematoxylin and eosin (H and E) staining. The Expression of HO-1 was detected by real-time RT-PCR, immunohistochemical techniques and western blotting.

RESULTSSerum ALT levels in every prevention group was lower than the group B [(149.44+/-16.51), (136.88+/-10.07), (127.38+/-11.03) vs (203.25+/-31.62) U/L, F = 92.498, P < 0.05], the SOD of liver homogenate in prevention group was significantly higher and the MDA was lower compared with the group B [SOD: (1.36+/-0.09), (1.42+/-0.13), (1.50+/-0.15) vs (1.08+/-0.19) U/mg, F = 13.671, P < 0.05; MDA: (0.294+/-0.026), (0.285+/-0.025), (0.284+/-0.028) vs (0.335+/-0.056) nmol/mg, F = 20.809, P < 0.05]. The pathological stages of hepatic fibrosis were all significantly reduced in prevention group (Chi2 test = 24.956, P < 0.05). Compared with group A, the mRNA and protein expressions of HO-1 were elevated (F = 4.549, 22.926, P < 0.05) in group B and increased in group C-E, but there is no significant difference existed.

CONCLUSIONBlueberry may have preventive and protective effects on CCl4-induced hepatic fibrosis by reducing lipid peroxidation. However, these effects may not be related to the activation of HO-1 during long-term of CCl4.

Animals ; Blueberry Plants ; chemistry ; Carbon Tetrachloride ; toxicity ; Drugs, Chinese Herbal ; pharmacology ; Heme Oxygenase (Decyclizing) ; blood ; Liver Cirrhosis, Experimental ; blood ; Male ; Malondialdehyde ; blood ; Rats ; Rats, Sprague-Dawley

BACKGROUNDVaccinium uliginosum L. is a type of blueberry found in the Chinese Changbai Mountains. We extracted Vaccinium uliginosum Anthocyanins (A(V.uli)) to investigate its bioactivity on suppressing cancer cells.

METHODSA(V.uli) was extracted under different conditions of temperature (10°C - 35°C), pH 1.0 - 3.0, and diatomaceous earth (1.0 g - 3.0 g), followed by a HPLC analysis for the determination of the ingredients. Its anticancer bioactivities on human colon and colorectal cancer cells (DLD-1 and COLO205) were compared with those on Lonicera caerulea Anthocyanins (A(L.cae)) and Vaccinium myrtillus Anthocyanins (A(V.myr)), using cell viability assays, DNA electrophoresis and nuclear morphology assays.

RESULTSThe optimum process of A(V.uli) extraction involved conditions of temperature 20°C, pH 2.0, and diatomaceous earth 1.0 g/50 g of fruit weight. A(V.uli) contained 5 main components: delphinidin (40.70 ± 1.72)%, cyanidin (3.40 ± 0.68)%, petunidin (17.70 ± 0.54)%, peonidin (2.90 ± 0.63)% and malvidin (35.50 ± 1.11)%. The malvidin percentage was significantly higher (P < 0.05) than it in A(V.myr). A(V.uli) complied with a dose-dependent repression of cancer cell proliferation with an IC(50) (50% inhibitory concentration) value of 50 µg/ml, and showed greater anticancer efficiency than A(L.cae) and A(V.myr) under the same cell treatment conditions. These observations were further supported by the results of nuclear assays.

CONCLUSIONSThe extraction protocol and conditions we used were effective for anthocyanin extraction. A(V.uli) could be a feasible practical research tool and a promising therapeutic source to suppress human colon or colorectal cancers.

Anthocyanins ; therapeutic use ; Antineoplastic Agents ; chemistry ; therapeutic use ; Apoptosis ; drug effects ; Blueberry Plants ; chemistry ; Cell Line, Tumor ; Cell Survival ; drug effects ; Chromatography, High Pressure Liquid ; DNA Fragmentation ; drug effects ; Humans ; Hydrogen-Ion Concentration ; Plant Extracts ; therapeutic use

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

OBJECTIVETo investigate the protective effect of blueberry extract (BE, 25% anthocyanins) against oxidative damage in primary cultures of rat hippocampal neurons induced by H2O2.

METHODSRat hippocampal neurons were randomly assigned to control group, H2O2 group and BE pretreatment groups, BE at six different doses (0.01, 0.1, 1.0, 10.0, 20.0 and 40 microg/ml) and then exposed to 50 micromol/L H2O2 for twenty-four hours. To selecte the most fittest concentration of BE by testing viability of neurons and activity of LDH. Then MDA concentration, SOD activity and neuronal apoptosis were(checked) measured.

RESULTS(1) Compared with H2O2 group, the hippocampal cell viabilities in the 0.1, 1.0 and 10 microg/ml BE groups were significantly increased from 57.44% to 78.42%, 87.71% and 72.40% separately. The activity of LDH in BE groups at varied concentrations (0.1, 1.0 and 10 microg/ml) was significiantly lower than that in H2O2 group. It was found that 1 microg/ml BE had the furthest protective effect against oxidative damage in primary cultures of rat hippocampal neurons induced by H2O2. (2) The concentration of MDA and the rate of neuronal apoptosis of BE group (1 microg/ml) were much lower than H2O2 group, while SOD activity was much higher.

CONCLUSIONProper dose of BE has remarkable protective effect against oxidative stress in primary cultures of rat hippocampal neurons induced by H2O2, the mechanism may be related to decreasing the neuronal apoptosis and enhancing the antioxidation of hippocampal neurons.

Animals ; Animals, Newborn ; Antioxidants ; pharmacology ; Blueberry Plants ; chemistry ; Cells, Cultured ; Hippocampus ; cytology ; Hydrogen Peroxide ; toxicity ; Neurons ; cytology ; drug effects ; Oxidative Stress ; drug effects ; Plant Extracts ; pharmacology ; Protective Agents ; pharmacology ; Rats ; Rats, Wistar

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*

The bioactive principles contained in blueberries (Vaccinium) are various kind of anthocyanins (anthocyanidins, or phenolic aglycone, conjugated with sugar), chlorogenic acid, flavonids, alpha-linolenic acid, pterostilbene, resveratrol, and vitamins. After oral administration, anthocyanins can pass through blood-brain barrier and thus appear in various organs and brain. Improve visual function by increasing rhodopsin regeneration and ocular health is the earliest reported bioactivities of anthocyanin. Recent studies demonstrated the benefit of blueberries to prevent the age-related chronic diseases such as cancer, diabeties, hyperlipidemia, hypertension, neurodegeneration, obesity, and osteoporosis through its apoptosis, antioxidant, antiinflammation, and antiangiogenesis effects. Blueberries can eradicate microorganisms for the prevention of symptomatic urinary tract infections in women. Thus, blueberries are recognized as one of the most nutritious foods and cultivated worldwide. However, how to prolong the shelving time of fresh fruit, well utilize the leaf and stem to isolate the bioactive chemicals, improve quality consistency of juicy and dry products, all should be further concerned.
Aging ; drug effects ; Animals ; Anthocyanins ; isolation & purification ; pharmacokinetics ; pharmacology ; Anti-Inflammatory Agents ; pharmacology ; Antihypertensive Agents ; pharmacology ; Antineoplastic Agents, Phytogenic ; pharmacology ; Antioxidants ; pharmacology ; Blood-Brain Barrier ; Blueberry Plants ; chemistry ; Chlorogenic Acid ; isolation & purification ; pharmacology ; Humans ; Hypoglycemic Agents ; pharmacology ; Hypolipidemic Agents ; pharmacology ; Plant Preparations ; pharmacology ; Plants, Edible ; chemistry