Effect of total flavonoids of buckwheat flower and leaf on myocardial cell apoptosis and Wnt/β-catenin/PPARγ pathway in arrhythmic rats / 中国中药杂志

This paper aimed to investigate the effect of total flavonoids of buckwheat flower and leaf on myocardial cell apoptosis and Wnt/β-catenin/peroxisome proliferator-activated receptor γ(PPARγ) pathway in arrhythmic rats. SD rats were randomly divided into a control group, a model group, a low-dose(20 mg·kg~(-1)) group of total flavonoids of buckwheat flower and leaf, a medium-dose(40 mg·kg~(-1)) group of total flavonoids of buckwheat flower and leaf, a high-dose(80 mg·kg~(-1)) group of total flavonoids of buckwheat flower and leaf, a propranolol hydrochloride(2 mg·kg~(-1)) group, with 12 rats in each group. Except the control group, rats in other groups were prepared as models of arrhythmia by sublingual injection of 1 mL·kg~(-1) of 0.002% aconitine. After grouping and intervention with drugs, the arrhythmia, myocardial cells apoptosis, myocardial tissue glutathione peroxidase(GSH-Px), catalase(CAT), malondialdehyde(MDA), serum interleukin-6(IL-6), prostaglandin E2(PGE2) levels, myocardial tissue apoptosis, and Wnt/β-catenin/PPARγ pathway-related protein expression of rats in each group were measured. As compared with the control group, the arrhythmia score, the number of ventricular premature beats, ventricular fibrillation duration, myocardial cell apoptosis rate, MDA levels in myocardial tissues, serum IL-6 and PGE2 levels, Bax in myocardial tissues, and Wnt1 and β-catenin protein expression levels increased significantly in the model group, whereas the GSH-Px and CAT levels, and Bcl-2 and PPARγ protein expression levels in myocardial tissues reduced significantly. As compared with the model group, the arrhythmia score, the number of ventricular premature beats, ventricular fibrillation duration, myocardial cell apoptosis rate, MDA leve in myocardial tissues, serum IL-6 and PGE2 levels, Bax in myocardial tissues, and Wnt1 and β-catenin protein expression levels reduced in the drug intervention groups, whereas the GSH-Px and CAT levels and Bcl-2 and PPARγ protein expression levels in myocardial tissues increased. The groups of total flavonoids of buckwheat flower and leaf were in a dose-dependent manner. There was no significant difference in the levels of each index in rats between the propranolol hydrochloride group and the high-dose group of total flavonoids of buckwheat flower and leaf. The total flavonoids of buckwheat flower and leaf inhibit the activation of Wnt/β-catenin pathway, up-regulate the expression of PPARγ, reduce oxidative stress and inflammatory damage in myocardial tissues of arrhythmic rats, reduce myocardial cell apoptosis, and improve the symptoms of arrhythmia in rats.

Effect of Tartary Buckwheat Boiled Noodles on Postprandial Blood Glucose Level and Its Active Components / 日本補完代替医療学会誌

Dry solid matter (rutin content: 51.6 mg/g; quercetin content: 72.2 mg/g) extracted from Tartary buckwheat boiled noodles using 70% methanol as the solvent was found to have α-glucosidase inhibitory activity. As for fractions fractionated by silica gel column chromatography, the fractions rich in quercetin and rutin showed remarkable α-glucosidase inhibitory activity. Tartary buckwheat boiled noodles used as samples in this study contained quercetin produced from rutin by the action of rutinase, suggesting that both rutin and quercetin contained were involved in the α-glucosidase inhibitory activity of the dry solid extract. Changes in postprandial blood glucose levels were compared for boiled noodles made from two types of buckwheat (i.e., Tartary buckwheat and common buckwheat), revealing that blood glucose elevation after eating Tartary buckwheat boiled noodles was suppressed. The blood glucose level 40 minutes after eating Tartary buckwheat boiled noodles was significantly low (p<0.05). It can be concluded that this might be caused by the α-glucosidase inhibitory activity of rutin (270.0 mg) and quercetin (330.5 mg), which correspond to a total amount of 935 mg of rutin equivalents, in the gastrointestinal tract. As a result, the digestion of carbohydrates contained in the samples consumed and their absorption by the intestine might be inhibited, resulting in the suppression of increases in blood glucose levels. The presence of a certain amount of quercetin was considered to be key to the suppression of blood glucose elevation. It is important to control rapid postprandial blood glucose increases to prevent diabetes from developing or becoming serious. This study suggests the potential for Tartary buckwheat boiled noodles to contribute to diabetes prevention.

Analysis of Flavonoid Content and Key Gene Expressions in Sprout and Seedling of Tartary Buckwheat / 中国实验方剂学杂志

Objective:To reveal the dynamic changes of flavonoids secondary metabolites and relevant genes expressions in the process of germination of tartary buckwheat seeds by investigating the content of catechins，epicatechins，rutin，and quercetin，and the expressions of their relevant genes in tartary buckwheat sprouts and seedlings，in order to provide scientific basis for the selection of high-quality, high-nutrition tartary buckwheat sprouts.Method:Contents of catechin，epicatechin，rutin，and quercetin in tartary buckwheat sprouts and seedlings were detected by UPLC-ESI-QQQ-MS，and the expression levels of genes relating to flavonoids synthesis in tartary buckwheat sprouts and seedlings were detected by real-time quantitative PCR.Result:There were differences between tartary buckwheat sprouts and seedlings in the relative contents of catechin，epicatechin，rutin and quercetin，as well as the expressions of relevant genes in the synthesis pathway, including FtPAL，FtC4H，Ft4CL，FtCHS，FtCHI，FtF3H，FtF3'H，FtFLS，FtDFR，FtLAR，FtANS，FtANR. The contents of flavonoids and the expressions of relevant genes in tartary buckwheat sprouts were higher than those in tartary buckwheat seedlings.Conclusion:The higher accumulation of secondary metabolites and flavonoids in tartary buckwheat sprouts may be related to tartary buckwheat seeds' resistance to the external environment in the initial growth stage of germination. From the perspective of application，there are more flavonoids in tartary buckwheat sprouts than in tartary buckwheat seedlings, indicating that tartary buckwheat sprouts have a higher nutritional value.

Characterization of the transcriptional profiles in common buckwheat (Fagopyrum esculentum) under PEG-mediated drought stress

BACKGROUND: Common buckwheat (Fagopyrum esculentum) is an important staple food crop in southwest China, where drought stress is one of the largest limiting factors that lead to decreased crop production. To reveal the molecular mechanism of common buckwheat in response to drought stress, we performed a comprehensive transcriptomics study to evaluate gene expression profiles of common buckwheat during PEG-mediated drought treatment. RESULTS: In total, 45 million clean reads were assembled into 53,404 unigenes with an average length of 749 bp and N50 length of 1296 bp. A total of 1329 differentially expressed genes (DEGs) were identified by comparing wellwatered and drought-treated plants, out of which 666 were upregulated and 663 were downregulated. Furthermore, we defined the functional characteristics of DEGs using GO and KEGG classifications. GO enrichment analysis showed that the DEGs were significantly overrepresented in four categories, namely, "oxidoreductase activity," "oxidation­reduction process," "xyloglucan:xyloglucosyl transferase activity," and "apoplast." Using KEGG pathway analysis, a large number of annotated genes were overrepresented in terms such as "plant hormone signal transduction," "phenylpropanoid biosynthesis," "photosynthesis," and "carbon metabolism." Conclusions: These results can be further exploited to investigate the molecular mechanism of common buckwheat in response to drought treatment and could supply with valuable molecular sources for abiotic-tolerant elite breeding programs in the future.

Influence of extract from fermented buckwheat flower and leaf in kidney injury in type 2 diabetic db/db mice / 吉林大学学报(医学版)

Objective: To discuss the protective effects of extract from fermented buckwheat flower and leaf (EFBFL) on the kidney injury in the spontaneously obese type II diabetic db/db mice, and to elucidate their possible action mechanisms from the protein expression levels of peroxisome proliferator-activated receptor y (PPARy) and nuclear factor-κB (NF-κB) in kidney tissue. Methods: The 8 weeks old male db/db mice were selected and the littermate db/m mice were used as normal control group; the db/db mice were randomly divided into model group, low dose of EFBFL group and high dose of EFBFL group, 10 mice in each group. The mice in low and high doses of EFBFL groups were intragastrically given 50 and 100 mg · kg: EFBFL, and the mice in normal control group and model group were intragastrically given the equal amount of distilled water once daily for 8 weeks accordingly. The levels of fasting blood-glucose (FBG) of the mice in various groups were tested respectively before and after administration. Full automatic biochemical analyzer was used to detect the levels of serum creatinine (SCr) and blood urea nitrogen (BUN) of the mice, and the kidney index was calculated. The morphology of kidney tissue was observed by HE staining and Masson staining, and immunohistochemical method and Western blotting method were used to detect the expression levels of PPARγ and NF-κB protein in kidney tissue. Results: Compared with model group, the levels of FBG and the levels of serum SCr and BUN in EFBFL groups were significantly decreased (P<0. 05 or P<0. 01), and the kidney indexes were increased (P<0. 01). Compared with normal control group, the kidney glomerular of the mice in model group was weaked, glomerular basement membrane was diffusely thickened, the foot process was coalesced or disappeared, and the kidney tissue fibrosis was serious; compared with the model group, the above performance of the mice in EFBFL groups were improved in different degrees, especially in high dose of EFBFL group. Compared with model group, the expression levels of PPARγ in kidney tissue of the mice in EFBFL groups were increased (P<0. 01) and the expression levels of NF-κB in kidney tissue of the mice were decreased (P<0. 01). Conclusion: EFBFL can improve the kidney injury in the spontaneously obese type II diabetic db/db mice, and its possible mechanism may be related to lowering the level of blood glucose, up-regulating the expression of PPARγ and down-regulating the expression of NF-κB in the kidney tissue of the mice.

Influence of extract from fermented buckwheat flower and leaf in kidney injury in type 2 diabetic db/db mice / 吉林大学学报(医学版)

Objective:To discuss the protective effects of extract from fermented buckwheat flower and leaf (EFBFL) on the kidney injury in the spontaneously obese type Ⅱ diabetic db/db mice,and to elucidate their possible action mechanisms from the protein expression levels of peroxisome proliferator-activated receptor γ(PPARγ) and nuclear factor-κB (NF-κB) in kidney tissue.Methods:The 8 weeks old male db/db mice were selected and the littermate db/m mice were used as normal control group;the db/db mice were randomly divided into model group,low dose of EFBFL group and high dose of EFBFL group,10 mice in each group.The mice in low and high doses of EFBFL groups were intragastrically given 50 and 100 mg · kg-1 EFBFL,and the mice in normal control group and model group were intragastrically given the equal amount of distilled water once daily for 8 weeks accordingly.The levels of fasting blood-glucose (FBG) of the mice in various groups were tested respectively before and after administration.Full automatic biochemical analyzer was used to detect the levels of serum creatinine (SCr) and blood urea nitrogen (BUN) of the mice,and the kidney index was calculated.The morphology of kidney tissue was observed by HE staining and Masson staining,and immunohistochemical method and Western blotting method were used to detect the expression levels of PPARγ and NF-κB protein in kidney tissue.Results:Compared with model group,the levels of FBG and the levels of serum SCr and BUN in EFBFL groups were significantly decreased (P＜0.05 or P＜0.01),and the kidney indexes were increased (P＜0.01).Compared with normal control group,the kidney glomerular of the mice in model group was weaked,glomerular basement membrane was diffusely thickened,the foot process was coalesced or disappeared,and the kidney tissue fibrosis was serious;compared with the model group,the above performance of the mice in EFBFL groups were improved in different degrees,especially in high dose of EFBFL group.Compared with model group,the expression levels of PPARγ in kidney tissue of the mice in EFBFL groups were increased (P＜0.01) and the expression levels of NF-κB in kidney tissue of the mice were decreased (P＜0.01).Conclusion:EFBFL can improve the kidney injury in the spontaneously obese type Ⅱ diabetic db/db mice,and its possible mechanism may be related to lowering the level of blood glucose,up-regulating the expression of PPARγ and down-regulating the expression of NF-κB in the kidney tissue of the mice.

Monoclonal Antibodies to Recombinant Fag e 3 Buckwheat Allergen and Development of a Two-site ELISA for Its Quantification

PURPOSE: Buckwheat is a major cause of anaphylaxis, and Fag e 3 is the key major allergen in buckwheat. However, an immunoassay system for the quantification of Fag e 3 has yet to be developed. METHODS: We developed a 2-site enzyme-linked immunosorbent assay (ELISA) using monoclonal antibodies (mAbs) produced against recombinant Fag e 3. We applied this ELISA to quantify native Fag e 3 in total buckwheat extract. RESULTS: Four clones of mAbs were produced, and all recognized vicilin allergens not only from buckwheat, but also from peanut and walnut. However, the ELISA using these antibodies was only able to quantify Fag e 3 in the total extract after addition of 1% sodium dodecyl sulphate (SDS) and heating, which facilitated dissociation of the allergen. The detection limit of the developed 2-site ELISA was 0.8 µg/mL. The measurement of Fag e 3 in the total extract of buckwheat showed that approximately 12% of protein in total buckwheat extract was Fag e 3. CONCLUSIONS: We have developed an ELISA system for the quantification of the group 3 buckwheat allergen, Fag e 3, specifically. This assay will be useful for standardization of buckwheat allergens and monitoring of buckwheat contamination in foods.

Hangover relieving effect of Sanghwang mushroom mycelium cultured in germinated buckwheat

The present study was performed to evaluate the hangover relieving effect of germinated buckwheat (GB) and Sanghwang mushroom mycelium cultured in GB (SGB). Both GB and SGB showed 1,1-diphenyl-2-picrylhydrazyl radical scavenging activities and significantly increased (p < 0.001) aldehyde dehydrogenase (ALDH) activities; up to 140% increase at concentrations of 16 µL/mL. Locomotor activity test results from alcohol-SGB and alcohol-GB groups showed improved motor activities over that of the alcohol-water group at 90 min post-administration. Both alcohol-GB and alcohol-SGB groups had significantly reduced (p < 0.001) alcohol (40.02 ± 33.38 µg/mL, 66.01 ± 22.04 µg/mL, respectively) and aldehyde (5.72 ± 0.47 µg/mL, 6.72 ± 1.70 µg/mL, respectively) concentrations in blood compared to those in the alcohol-water group (199.75 ± 33.83 µg/mL, 50.43 ± 13.88 µg/mL, respectively) at 90 min post-administration. Based on cDNA microarray analysis, expressions of ALDH genes ALDH1a7 and ALDH18a1 and cytochrome P450 (CY450) gene CYP4a30b were upregulated in the alcohol-GB and alcohol-SGB groups compared to levels in the control group. Overall, the results suggest that both GB and SGB have hangover relieving effects by reducing blood acetaldehyde levels. The molecular mechanisms may involve ALDH activation and upregulated expression of alcohol metabolism-related genes such as ALDH and CYP450.

Purification, characterization and partial primary structure analysis of rutin-degrading enzyme in tartary buckwheat seeds / 生物工程学报

Rutin-degrading enzymes (RDE) can degrade rutin into poorly water soluble compound, quercetin, and cause the bitter taste in tartary buckwheat. In the present study RDE from Yu 6-21 tartary buckwheat seeds was purified by ammonium sulphate precipitation, followed by hydrophobic interaction chromatography on Phenyl Sepharose CL-4B, ion exchange chromatography on CM-Cellulose and gel filtration chromatography on Sephadex G-150. Purified RDE showed single band with molecular weight of 66 kDa on SDS-PAGE. The optimum pH and temperature of RDE were 5.0 and 50 ℃ respectively. The Km was 0.27 mmol/L, and the Vmax was 39.68 U/mg. The RDE activity could be inhibited by Cu²⁺, Zn²⁺, Mn²⁺ and EDTA, and showed tolerance to 50% methanol (V/V). The N terminal sequence (TVSRSSFPDGFLFGL) was obtained by Edman degradation method and 15 internal peptide sequences were determined by MALDI-TOF-MS (matrix-assisted laser desorption ionization time of flight mass spectrometry). These results established the foundations for identification of the candidate gene of RDE via transcriptome data and further studying RDE biological function.

Effects of extract from fermented buckwheat flower and leaf(EFBFL)on myocardial injury in type 2 diabetic db/db mice / 中国药理学通报

Aim To discuss the effectsof extract from fermented buckwheat flower and leaf(EFBFL) on myocardial injury in spontaneously obese type Ⅱ diabetic db/db mice and its mechanism.Methods 9-week-old male db/db mice were randomly divided into high level EFBFL dose group(EFBFL-H, 0.1 g·kg-1), low level EFBFL dose group(EFBFL-L, 0.05 g·kg-1),metformin hydrochloridecontrol group, model control group, and normal control group, with 10 mice in each group.All groups were treated with 8 wks of drugs by gastric perfusion.The random blood glucose(RBG) was tested respectively at the end of 2nd, 4th, 6th, and 8th wk.Finally, the levels of creatine kinase(CK) creatine kinase MB(CK-MB), andadvanced glycosylation endproducts(AGEs) were detected after 8 wks.The morphological changes of myocardium were observed under light microscope by HE staining, and the ultrastructure of myocardium was observed under electron microscope.Immunohistochemical method and Western blot were used to detect myocardial tissue glucose transporter-4(Glut-4).Results EFBFLcould repress patho-proceeding of myocardial fibrosis efficiently, and significantly decrease the level of blood glucose, CK,CK-MB, and AGEs in db/db mice.Meanwhile, it could increase the expression of Glut-4 in myocardial tissues of mice.Conclusions EFBFL can prevent myocardial injury in spontaneously obese type Ⅱ diabetic db/db mice.The possible mechanism may be related to lowering the level of blood glucose and serum AGEs and up-regulating Glut4of cardiac muscle.

Clinical and laboratory findings of childhood buckwheat allergy in a single tertiary hospital / 소아과

PURPOSE: Buckwheat allergy is one of the most severe types of food allergy in some countries, especially among children. However, few studies have investigated this condition. The aim of this study was to report the clinical and laboratory findings in Korean children with buckwheat allergy. METHODS: Thirty-seven subjects, aged 1 to 14 years, were enrolled by retrospective medical record review from January 2000 through May 2015 at the Department of Pediatrics in Ajou University Hospital. The demographic profile, previous exposure to buckwheat pillows, clinical symptoms, and laboratory findings of each subject were recorded. RESULTS: Twenty-six of the 37 children had immediate-type allergic symptoms to buckwheat, while 11 subjects were tolerant to buckwheat. Seventeen out of 26 buckwheat allergic children (65.4%) had anaphylaxis. The median buckwheat specific IgE level in the buckwheat allergic group (7.71 kU(A)/L) was significantly higher (P<0.001) than in the buckwheat tolerant group (0.08 kU(A)/L) with an optimal cutoff value of 1.27 kU(A)/L (sensitivity 84.6%, specificity 100%). When adjusted for age, the difference between the 2 groups showed no statistical significance (P=0.063). In subjects who had anaphylaxis, buckwheat-specific IgE levels ranged from 0.37 to 100 kUA/L. CONCLUSION: Almost two-thirds of buckwheat-allergic children had anaphylaxis, and a wide-range of buckwheat specific IgE levels were observed in these children. Anaphylaxis occurred in a subject with a remarkably low IgE level (0.37 kU(A)/L).

Protective effect of total flavones of buckwheat flowers and leaves on myocardial fibrosis of type 2 diabetic rats and its mechanism / 吉林大学学报(医学版)

Objective: To study the protective effect of the total flavones of buckwheat flowers and leaves (TFBFL)on the myocardial injury of the type 2 diabetic rats,and to clarify its mechanism.Methods:The diabetic rat models were established by intraperitoneally injecting streptozotocin (STZ)and taking high-fat diet.The model rats were divided into diabetic model (DM)group and TFBFL treatment (TFBFL)group,while normal control group was set up, 10 rats per group.The rats in TFBFL group were administered with 200 mL· kg-1 · d-1 TFBFL,while the rats in other two groups were given with normal water (10 mL·kg-1 ·d-1 )instead.The rats in various groups was administered once a day for 8 weeks.The body weight (BW),heart weight index (HWI), level of fasting blood glucose (FBG)and cardiac function indexes (HR,LVSP,LVEDP and ± dp/dtmax )of the rats in various groups were measured.The myocardial tissue morphology of the rats in various groups was evaluated by electron microscope and the collagen levels in myocardium tissue of the rats in various groups were observed by Masson staining.The expression levels of TGF-β1 protein in myocardium tissue of the rats in various groups were detected by Western blotting method.Results:Compared with normal control group,the HWI,FBG and LVEDP levels of the rats in DM group were increased (P 0.05).The Masson staining results showed the morphology of myocardium tissue of the rats in TFBFL group was improved and the collagen level was decreased. The Western blotting results suggested that the TGF-β1 protein expression level in myocardium tissue of the rats in DM group was significantly increased compared with normal control group (P < 0.05),and the TGF-β1 protein expression level of the rats in TFBFL group was decreased compared with DM group (P <0.05).Conclusion:TFBFL exerts a certain protective effect on the myocardial fibrosis of diabetic rats through inhibiting the TGF-β1 expression in myocardium tissue.

Significance of 40-, 45-, and 48-kDa Proteins in the Moderate-to-Severe Clinical Symptoms of Buckwheat Allergy

PURPOSE: This study was aimed to investigate the relationship between the allergen components and moderate-to-severe allergic reactions in patients with buckwheat allergy. METHODS: Fifteen patients with a history of buckwheat ingestion and a buckwheat specific IgE level> or =0.35 kU/L were enrolled. They were divided into 2 groups according to clinical severity scores, with 0-1 being asymptomatic-to-mild and 2-4 being moderate-to-severe symptoms. Immunoblotting was performed to investigate IgE reactivity toward buckwheat allergens and to measure intensity of each component by using a reflective densitometer. RESULTS: The proportions of positive band to the 16 kDa (62.5% vs 0%, P=0.026) and 40-50 kDa (87.5% vs 28.6%, P=0.041) buckwheat allergens in the grade 2-4 group were higher than those in grade 0-1 group. The level of buckwheat specific IgE of grade 2-4 group was higher than that of grade 0-1 group (41.3 kU/L vs 5.5 kU/L, P=0.037). The median optical densities (ODs) of IgE antibody binding to 40-50 kDa protein were higher in the grade 2-4 group, compared with those in the grade 0-1 group (130% OD vs 60.8% OD, P=0.037). CONCLUSIONS: The 40-50 kDa protein is implicated as an important allergen to predict moderate-to-severe clinical symptoms in Korean children with buckwheat allergy.

Study on Anti-inflammatory Function of Gold Buckwheat Fermented Tea / 世界科学技术-中医药现代化

This study was aimed to observe the effect of gold buckwheat fermented tea on both acute and chronic inflammation in mice. Ear swelling was induced by xylene. Toe swelling was induced by carrageenan. The acute inflammation model was induced by the increasing of capillary permeability in the abdomen of mice by acetic acid. The chronic inflammation model of granuloma was used in the evaluation of anti-inflammatory function of gold buckwheat fermented tea. The results showed that the gold buckwheat fermented tea can obviously decrease the rate of xylene-induced ear swelling and the degree of carrageenan-induced toe swelling in mice. And the gold buckwheat fermented tea had antagonism effect on acute inflammation caused by acetic acid through the increasing of capillary permeability in abdomen of mice. Furthermore, the gold buckwheat fermented tea had obvious inhibition effects on proliferation of granuloma in chronic inflammation. It was concluded that the gold buckwheat fermented tea had the prevention and treatment effect on both acute and chronic inflammation.

Effect of chlorocholine chlorid on phenolic acids accumulation and polyphenols formation of buckwheat plants

BACKGROUND: Effect of chlorocholine chloride (CCC) on phenolic acids composition and polyphenols accumulation in various anatomical parts (stems, leaves and inflorescences) of common buckwheat (Fagopyrum esculentum Moench) in the early stages of vegetation period were surveyed. RESULTS: Treatment of buckwheat seeds with 2% of CCC has been increased content of total phenolics in the stems, leaves and inflorescences. On analyzing the different parts of buckwheat plants, 9 different phenolic acids - vanilic acid, ferulic acid, trans-ferulic acid, chlorogenic acid, salycilic acid, cinamic acid, p-coumaric acid, p-anisic acid, methoxycinamic acid and catechins were identified. The levels of identified phenolic acids varied not only significantly among the plant organs but also between early stages of vegetation period. Same changes as in contents of chlorogenic acid, ferulic acid, trans-ferulic acid were found for content of salycilic acid. The content of these phenolic acids has been significant increased under effect of 2% CCC treatment at the phase I (formation of buds) in the stems and at the phase II (beginning of flowering) in the leaves and then inflorescences respectively. The content of catechins as potential buckwheat antioxidants has been increased at the early stages of vegetation period after treatment with 2% CCC. CONCLUSIONS: The obtained results suggest that influence of CCC on the phenolics composition can be a result of various mechanisms of CCC uptake, transforming and/or its translocation in the buckwheat seedlings.

Effects of enzymatic hydrolysis of buckwheat protein on antigenicity and allergenicity

BACKGROUND/OBJECTIVES: Due to its beneficial health effects, use of buckwheat has shown a continuous increase, and concerns regarding the allergic property of buckwheat have also increased. This study was conducted for evaluation of the hydrolytic effects of seven commercial proteases on buckwheat allergens and its allergenicity. MATERIALS/METHODS: Extracted buckwheat protein was hydrolyzed by seven proteolytic enzymes at individual optimum temperature and pH for four hours. Analysis was then performed using SDS-PAGE, immunoblotting, and competitive inhibition ELISA (ciELISA) with rabbit antiserum to buckwheat protein, and direct ELISA with pooled serum of 21 buckwheat-sensitive patients. RESULTS: Alkaline protease, classified as serine peptidase, was most effective in reducing allergenicity of buckwheat protein. It caused decomposition of the whole buckwheat protein, as shown on SDS-PAGE, and results of immunoblotting showed that the rabbit antiserum to buckwheat protein no longer recognized it as an antigen. Allergenicity showed a decrease of more than 50% when pooled serum of patients was used in ELISA. Two proteolytic enzymes from Aspergillus sp. could not hydrolyze buckwheat allergens effectively, and the allergenicity even appeared to increase. CONCLUSIONS: Serine-type peptidases appeared to show a relatively effective reduction of buckwheat allergenicity. However, the antigenicity measured using rabbit antiserum did not correspond to the allergenicity measured using sera from human patients. Production of less allergenic buckwheat protein may be possible using enzymatic hydrolysis.

Effect of tartary buckwheat flavonoids on neurological function and expression of cerebral nitric oxide synthases in rats with focal cerebral ischemia-reperfusion / 中国药学杂志

OBJECTIVE: To investigate the protective effect of the decoction of tartary buckwheat flavonoids on focal cerebral is-chemia-reperfusion injury of rats. MEHTODS: Fifty male SD rats will be randomly divided into normal group, cerebral ischemia-reperfusion group (model group), high dose of tartary buckwheat flavonoids group, middle dose of tartary buckwheat flavonoids group and low dose of tartary buckwheat flavonoids group, each groups of 10 rats. A rat model of the right middle cerebral artery occlusion/reper-fusion(MCAO) was established by the filament method. After being operated, treatment-group rats will be administered 100,75 and 50 mg · kg-1 of the decoction of tartary buckwheat flavonoids three times a day for 7 consecutive days, after administrated for 7 d, rats in each group will undergo neurobehavioral tests. Expressions of cerebral inducible nitric oxide synthase (iNOS) and endothelial nitric oxide synthase (eNOS) were measured by SP immunohistochemistry. The optical density value (OD) was measured by imaging analysis, and the percentage of cells with iNOS and eNOS positive expression was analyzed under light microscope. RESULTS: Compared with ischemia-reperfusion group, neurological function score increased in the decoction of tartary buckwheat flavonoids groups. Treatment groups had lower expression level of iNOS but higher expression level of eNOS than those in the model groups (P < 0.01, P < 0.05). The number of neurons of Hippocampal CA1 was increased (P < 0.05). CONCLUSION: Tartary buckwheat flavonoids can improve neurological function and decrease the expression of iNOS and increase the expression of eNOS in cerebral ischemia-reperfusion rats, which may contribute to the protection of neural function.

Allergenic potential and enzymatic resistance of buckwheat

Buckwheat is known as a health food but is one of the major food allergens triggering potentially fatal anaphylaxis in Asia, especially in Japan and Korea. This study was conducted to investigate the characteristic of enzymatic resistance of buckwheat protein and allergenic potential. Enzymatic resistance of buckwheat protein was performed with in vitro digestibility test in simulated gastric fluid (SGF), pH 1.2, using pepsin and simulated intestinal fluid (SIF) using chymotrypsin. Reactivity of buckwheat proteins to human IgE was performed using six allergic patients sensitized to buckwheat. Buckwheat's IgE levels were measured using the Phadia UniCAP-system. Buckwheat protein, 16 kDa, still remained after 30 min treatment of pepsin on SDS-PAGE. Even though 16 kDa almost disappeared after 60 min treatment, two out of the six buckwheat patients' sera showed reactivity to hydrolysate after 60 min treatment, indicating that allergenicity still remained. In simulated intestinal fluid (SIF) using chymotrypsin, buckwheat protein, 24 kDa, showed resistance to hydrolysis with chymotrypsin on SDS-PAGE, and still had allergenicity based on the result of ELISA. Our results suggest that buckwheat proteins have strong resistance to enzyme degradation. This may be attributed in part to the allergenic potential of buckwheat. Further study should be continued regarding buckwheat allergy.

Effects of adlay, buckwheat, and barley on transit time and the antioxidative system in obesity induced rats

In the present study, we examined whether four grains including adlay (AD), buckwheat (BW), glutinous barley (GB), and white rice (WR) affect the duration of food residence in the gastrointestinal tract and hepatic enzyme activities in rats fed different combinations of the grains. The rats were raised for 4 weeks on a high fat diet based on the American Institute of Nutrition-93 (AIN-93G) diets containing 1% cholesterol and 20% dietary lipids. Forty male rats were divided into four groups and raised for 4 weeks with a diet containing one of the grains. Corresponding to the dietary fiber contents of the experimental grains, gut transit time was shortest in the rats fed GB and increased in the order of BW, AD, and WR. In addition, the accumulated shortest transit time occurred in the GB group. Gut transit time affected weight gain and major organ weight, as it was closely related to the absorption of nutrients. The level of thiobarbituric acid reactive substance (TBARS) in liver was higher in rats fed WR, AD, BW, and GB, indicating that the other grains decreased oxidative stress in vivo more than WR. Glutathione, glutathione peroxidase, and glutathione S-transferase levels in the AD, BW, and GB groups were significantly higher than those in the WR group. In conclusion, reduced colonic transit time has been implicated in reducing the incidence of colon cancer, as evidenced by populations consuming diets rich in fiber. Whole grains such as AD, BW, and GB may contribute to a significant supply of antioxidants to prevent oxidative stress if they are consumed in large amounts.

Effect of Cosensitization with Buckwheat Flour Extract on the Production of House Dust Mite-specific IgE

There are studies reporting food sensitization in infancy increases the risk of sensitization to inhalants later in life. We performed a study to evaluate whether cosensitization with buckwheat (BW) has an effect on the production of house dust mite-IgE. C3H/HeJ mice (4 weeks, female) were sensitized with house dust mite (HDM)/Al (OH)3, intraperitoneally on day 0, followed by 4 intranasal sensitizations (on days 14, 15, 16, and 21). Group 1 was cosensitized intragastrically with BW/cholera toxin (CT) (on days 0, 1, 2, 7, and 18) during sensitization with HDM, group 2 was cosensitized intragastrically with CT only (on days 0, 1, 2, 7, and 18), and group 3 was used as controls. HDM- and BW-IgE and antigen-specific T-cell proliferation and cytokine production were evaluated. In Group 1, BW-IgE levels were highest at week 4, and the HDM-IgE at week 3 (98.45+/-64.37 ng/mL and 169.86+/-55.54 ng/mL, respectively). In Group 2, HDM-IgE levels reached a peak at week 3, remarkably higher (810.52+/-233.29 ng/mL) compared to those of Group 1 (169.86+/-55.54 ng/mL). The interleukin (IL)-4 and interferon (IFN)-gamma in the HDM-stimulated culture supernatants of splenocytes were not significantly different among groups. We postulate that the cosensitization with BW may down-regulate the specific IgE response to HDM.