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1.
Food Res Int ; 184: 114233, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38609217

RESUMO

Meju is essential for making diverse traditional fermented soybean foods in Korea. To understand the changes in carbohydrates during fermentation, we aimed to identify autochthonous microorganisms from spontaneously fermented meju and compare the alterations in monosaccharides and oligosaccharides throughout the fermentation process. Microbial diversity was determined using a metabarcoding approach, and monosaccharide and oligosaccharide profiles were obtained by HPLC-Q-TOF MS and HPLC-MS/MS analyses, respectively. The dominant bacterial genera were Weissella, Lactobacillus, and Leuconostoc, while Mucor was highly abundant in the fungal community. The total monosaccharide content increased from Day 0 to Day 50, with the highest amount being 4.37 mg/g. Oligosaccharide profiling revealed the degradation of soybean dietary fiber during fermentation, and novel oligosaccharide structures were also discovered. Correlation analysis revealed that the fungus Mucor was positively related to pentose-containing oligosaccharides, galactose, and galacturonic acid, indicating that Mucor may degrade soybean dietary fibers such as xylogalacturonan, arabinogalactan, and rhamnogalacturonan. The negative relationships between the abundances of Weissella and oligo- and monosaccharides suggested that the bacteria may utilize saccharides for fermentation. These findings provide insights into the mechanisms underlying carbohydrate degradation and utilization; the key components involved in saccharide transformation that contribute to the characteristics of traditional meju were subsequently identified.


Assuntos
Microbiota , Monossacarídeos , Glycine max , Fermentação , Espectrometria de Massas em Tandem , Oligossacarídeos , Fibras na Dieta
2.
Pharmaceuticals (Basel) ; 16(3)2023 Mar 07.
Artigo em Inglês | MEDLINE | ID: mdl-36986507

RESUMO

Abietic acid (AA) is known to have beneficial effects on inflammation, photoaging, osteoporosis, cancer, and obesity; however, its efficacy on atopic dermatitis (AD) has not been reported. We investigated the anti-AD effects of AA, which was newly isolated from rosin, in an AD model. To achieve this, AA was isolated from rosin under conditions optimized by response surface methodology (RSM), and its effects on cell death, iNOS-induced COX-2 mediated pathway, inflammatory cytokine transcription, and the histopathological skin structure were analyzed in 2,4-dinitrochlorobenzene (DNCB)-treated BALB/c mice after treatment with AA for 4 weeks. AA was isolated and purified through isomerization and reaction-crystallization under the condition (HCl, 2.49 mL; reflux extraction time, 61.7 min; ethanolamine, 7.35 mL) established by RSM, resulting in AA with a purity and extraction yield of 99.33% and 58.61%, respectively. AA exhibited high scavenging activity against DPPH, ABTS, and NO radicals as well as hyaluronidase activity in a dose-dependent manner. The anti-inflammatory effects of AA were verified in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages through amelioration of the inflammatory response, including NO production, iNOS-induced COX-2 mediated pathway activation, and cytokine transcription. In the DNCB-treated AD model, the skin phenotypes, dermatitis score, immune organ weight, and IgE concentration were significantly ameliorated in the AA cream (AAC)-spread groups compared to the vehicle-spread group. In addition, AAC spread ameliorated DNCB-induced deterioration of skin histopathological structure through the recovery of the thickness of the dermis and epidermis and the number of mast cells. Furthermore, activation of the iNOS-induced COX-2 mediated pathway and increased inflammatory cytokine transcription were ameliorated in the skin of the DNCB+AAC-treated group. Taken together, these results indicate that AA, newly isolated from rosin, exhibits anti-AD effects in DNCB-treated AD models, and has the potential to be developed as a treatment option for AD-related diseases.

3.
Foods ; 11(16)2022 Aug 21.
Artigo em Inglês | MEDLINE | ID: mdl-36010531

RESUMO

Sargassum thunbergii (Mertens ex Roth) Kuntze (ST) is a brown alga rich in indole-2-carboxaldehyde. This study aimed to investigate the anti-obesity effects of ethanol extract from ST in in vitro and in vivo models. In 3T3-L1 cells, ST extract significantly inhibited lipid accumulation in mature adipocytes while lowering adipogenic genes (C/epba and Pparg) and enhancing metabolic sensors (Ampk, Sirt1), thermogenic genes (Pgc-1a, Ucp1), and proteins (p-AMPK/AMPK and UCP1). During animal investigation, mice were administered a chow diet, a high-fat diet (HF), or an HF diet supplemented with ST extract (at dosages of 150 and 300 mg/kg bw per day) for 8 weeks (n = 10/group). ST extract administration decreased weight gain, white adipose tissue weight, LDL-cholesterol, and serum leptin levels while improving glucose intolerance. In addition, ST extract increased the expression of Ampk and Sirt1 in adipose tissue and in the liver, as well as p-AMPK/AMPK ratio in the liver, compared to HF-fed mice. The abundance of Bacteroides vulgatus and Faecalibacterium prausnitzii in the feces increased in response to ST extract administration, although levels of Romboutsia ilealis decreased compared with those in HF-fed mice. ST extract could prevent obesity in HF-fed mice via the modulation of AMPK activation and gut microbiota composition.

4.
J Microbiol Biotechnol ; 32(5): 594-601, 2022 May 28.
Artigo em Inglês | MEDLINE | ID: mdl-35484970

RESUMO

This study evaluated the biological properties of lemongrass (Cymbopogon citratus) extracts. The EtOAc extract of lemongrass had DPPH, TEAC, and nitric oxide-scavenging activity assay results of 58.06, 44.14, and 41.08% at the concentration of 50, 10, and 50 µg/ml, respectively. The EtOAc extract had higher elastase and collagenase inhibitory activities than the 80% MeOH, n-hexane, BuOH, and water extracts and comparable whitening activity toward monophenolase or diphenolase. Also, the EtOAc fraction had higher lipase inhibitory and antimicrobial activities against Cutibacterium acnes among extracts which is known to an important contributor to the progression of inflammatory acne vulgaris, and an opportunistic pathogen present in human skin. Total phenolic and flavonoid concentrations in the EtOAc extract were 132.31 mg CAE/g extract and 104.50 mg NE/g extract, respectively. Biologically active compounds in lemongrass extracts were analyzed by LC-MS. This study confirms that lemongrass extracts have potential use as cosmetic skincare ingredients. Thus, lemongrass can be considered a promising natural source of readily available, low-cost extracts rich in antioxidant, skincare, and antimicrobial compounds that might be suitable for replacing synthetic compounds in the cosmeceutical industry.


Assuntos
Acne Vulgar , Anti-Infecciosos , Cosméticos , Cymbopogon , Acne Vulgar/tratamento farmacológico , Acne Vulgar/microbiologia , Anti-Infecciosos/química , Anti-Infecciosos/farmacologia , Antioxidantes/química , Antioxidantes/farmacologia , Cymbopogon/química , Humanos , Extratos Vegetais/química , Extratos Vegetais/farmacologia
5.
Molecules ; 26(23)2021 Nov 28.
Artigo em Inglês | MEDLINE | ID: mdl-34885790

RESUMO

This study investigated the laxative effects of phlorotannins (Pt) derived from Ecklonia cava (E. cave) on chronic constipation by evaluating alterations in stool parameters, gastrointestinal motility, histopathological structure, mucin secretion, gastrointestinal hormones, muscarinic cholinergic regulation, and fecal microbiota in SD rats with loperamide (Lop)-induced constipation subjected to Pt treatment. Stool-related parameters (including stool number, weight, and water contents), gastrointestinal motility, and length of intestine were significantly enhanced in the Lop+Pt-treated group as compared to the Lop+Vehicle-treated group. A similar recovery was detected in the histopathological and cytological structure of the mid-colon of Lop+Pt-treated rats, although the level of mucin secretion remained constant. Moreover, rats with Lop-induced constipation subjected to Pt treatment showed significant improvements in water channel expression, gastrointestinal hormone secretions, and expression of muscarinic acetylcholine receptors M2/M3 (mAChRs M2/M3) and their mediators of muscarinic cholinergic regulation. Furthermore, the Lop+Pt-treated group showed a significant recovery of Bifidobacteriaceae, Muribaculaceae, Clostridiaceae, and Eubacteriaceae families in fecal microbiota. Taken together, these results provide the first evidence that exposure of SD rats with Lop-induced constipation to Pt improves the constipation phenotype through the regulation of membrane water channel expression, GI hormones, the mAChR signaling pathway, and fecal microbiota.


Assuntos
Constipação Intestinal/tratamento farmacológico , Laxantes/uso terapêutico , Phaeophyceae/química , Taninos/uso terapêutico , Animais , Constipação Intestinal/induzido quimicamente , Laxantes/química , Loperamida , Masculino , Extratos Vegetais/química , Extratos Vegetais/uso terapêutico , Ratos Sprague-Dawley , Taninos/química
6.
Food Sci Biotechnol ; 27(5): 1485-1492, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30319859

RESUMO

This study examined the biological functions of the butanol extracts of green pine cones (GPCs) that had not ripened completely. The butanol extracts of GPC showed 78.22% DPPH-scavenging activity, 53.55% TEAC and 71.50% hyaluronidase (HAase) inhibition activity. They also exhibited inhibition activity against food poisoning microorganisms. The contents of total phenolic compounds and total flavonoids were 296.75 and 26.07 mg/g, respectively. Biologically active compounds were analyzed and separated using HPLC related to the DPPH-scavenging and HAase inhibition activities. Gallotannin was the primary biologically active compound with DPPH-scavenging and HAase inhibition activities in the GPC butanol extracts.

7.
Int J Mol Med ; 40(5): 1365-1376, 2017 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-28949387

RESUMO

Asthma is a chronic inflammatory disease characterized by T-lymphocyte and eosinophil infiltration, mucus overproduction and airway hyper-responsiveness. The present study examined the therapeutic effects and action mechanism of a saponin-enriched extract of Asparagus cochinchinensis (SEAC) on airway inflammation and remodeling in an ovalbumin (OVA)-induced asthma model. To accomplish this, alterations of the nitric oxide (NO) level, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression levels, as well as variations in immune cell numbers, immunoglobulin E (IgE) concentration, histopathological structure and inflammatory cytokine levels were measured in lipopolysaccharide (LPS)-activated RAW264.7 cells or an OVA-induced mouse model of asthma treated with SEAC. The concentration of NO and mRNA levels of COX-2 and iNOS were significantly decreased in the SEAC + LPS-treated RAW264.7 cells compared with the vehicle + LPS-treated RAW264.7 cells. Additionally, in the OVA-induced asthma model, the number of immune cells in the bronchoalveolar lavage fluid, the concentration of OVA-specific IgE, the infiltration of inflammatory cells, the bronchial thickness and the levels of the inflammatory mediators interleukin-4 (IL-4), IL-13 and COX-2 were significantly lower in the OVA + SEAC­treated group compared with the OVA + vehicle­treated group. In addition, a significant reduction in goblet cell hyperplasia, peribronchiolar collagen layer thickness and VEGF expression for airway remodeling was detected in the OVA + SEAC­treated group compared with the OVA + vehicle­treated group. These findings indicate that SEAC is a suppressor of airway inflammation and remodeling, and may therefore be useful as an anti-inflammatory drug for the treatment of asthma.


Assuntos
Remodelação das Vias Aéreas/efeitos dos fármacos , Antiasmáticos/farmacologia , Anti-Inflamatórios/farmacologia , Asma/etiologia , Asma/patologia , Ovalbumina/efeitos adversos , Saponinas/farmacologia , Animais , Antiasmáticos/química , Anti-Inflamatórios/química , Antioxidantes/química , Antioxidantes/farmacologia , Asparagus/química , Asma/tratamento farmacológico , Asma/metabolismo , Biomarcadores , Líquido da Lavagem Broncoalveolar , Linhagem Celular , Ciclo-Oxigenase 2/metabolismo , Citocinas/genética , Citocinas/metabolismo , Modelos Animais de Doenças , Feminino , Imunoglobulina E/imunologia , Mediadores da Inflamação/metabolismo , Contagem de Leucócitos , Camundongos , Óxido Nítrico Sintase Tipo II/metabolismo , Extratos Vegetais , Células RAW 264.7 , Espécies Reativas de Oxigênio/metabolismo , Saponinas/química
8.
J Microbiol Biotechnol ; 27(4): 701-708, 2017 Apr 28.
Artigo em Inglês | MEDLINE | ID: mdl-28189140

RESUMO

This study was conducted to evaluate the hyaluronidase (HAase) inhibition activity of Asparagus cochinchinesis (AC) extracts following fermentation by Weissella cibaria through response surface methodology. To optimize the HAase inhibition activity, a central composite design was introduced based on four variables: the concentration of AC extract (X1: 1-5%), amount of starter culture (X2: 1-5%), pH (X3: 4-8), and fermentation time (X4: 0-10 days). The experimental data were fitted to quadratic regression equations, the accuracy of the equations was analyzed by ANOVA, and the regression coefficients for the surface quadratic model of HAase inhibition activity in the fermented AC extract were estimated by the F test and the corresponding p values. The HAase inhibition activity indicated that fermentation time was most significant among the parameters within the conditions tested. To validate the model, two different conditions among those generated by the Design Expert program were selected. Under both conditions, predicted and experimental data agreed well. Moreover, the content of protodioscin (a well-known compound related to anti-inflammation activity) was elevated after fermentation of the AC extract at the optimized fermentation condition.


Assuntos
Asparagus/enzimologia , Fermentação , Hialuronoglucosaminidase/antagonistas & inibidores , Hialuronoglucosaminidase/metabolismo , Extratos Vegetais/farmacologia , Weissella/metabolismo , Análise de Variância , Anti-Inflamatórios/farmacologia , Asparagus/química , Asparagus/microbiologia , Cromatografia Líquida de Alta Pressão/métodos , Diosgenina/análogos & derivados , Diosgenina/farmacologia , Concentração de Íons de Hidrogênio , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Saponinas/farmacologia , Weissella/crescimento & desenvolvimento
9.
J Microbiol Biotechnol ; 26(9): 1527-32, 2016 Sep 28.
Artigo em Inglês | MEDLINE | ID: mdl-27238939

RESUMO

Strain SPF4211, having hyaluronidase (HAase) inhibition activity, was isolated from P. davidiana (Carriere) Franch fruit (PrDF) sugar extract. The phenotypic and biochemical properties based on 16S rDNA sequencing and an API 50 CHB kit suggested that the organism was B. subtilis. To optimize the HAase inhibition activity of PrDF extract by fermentation of strain SPF4211, a central composite design (CCD) was introduced based on three variables: concentration of PrDF extract (X1: 1-5%), amount of starter culture (X2: 1-5%), and fermentation time (X3: 0-7 days). The experimental data were fitted with quadratic regression equations, and the accuracy of the equations was analyzed by ANOVA. The statistical model predicted the highest HAase inhibition activity of 37.936% under the optimal conditions of X1 = 1%, X2 = 2.53%, and X3 = 7 days. The optimized conditions were validated by observation of an actual HAase inhibition activity of 38.367% from extract of PrDF fermented by SPF4211. These results agree well with the predicted model value.


Assuntos
Bacillus subtilis/metabolismo , Frutas/enzimologia , Hialuronoglucosaminidase/antagonistas & inibidores , Hialuronoglucosaminidase/metabolismo , Extratos Vegetais/farmacologia , Prunus/enzimologia , Fermentação , Frutas/química , Frutas/metabolismo , Frutas/microbiologia , Prunus/química , Prunus/metabolismo , Prunus/microbiologia
10.
Lab Anim Res ; 32(1): 34-45, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-27051441

RESUMO

Asparagus cochinchinensis has been used to treat various diseases including fever, cough, kidney disease, breast cancer, inflammatory disease and brain disease, while IL-4 cytokine has been considered as key regulator on the skin homeostasis and the predisposition toward allergic skin inflammation. However, few studies have investigated its effects and IL-4 correlation on skin inflammation to date. To quantitatively evaluate the suppressive effects of ethyl acetate extracts of A. cochinchinensis (EaEAC) on phthalic anhydride (PA)-induced skin inflammation and investigate the role of IL-4 during their action mechanism, alterations in general phenotype biomarkers and luciferase-derived signals were measured in IL-4/Luc/CNS-1 transgenic (Tg) mice with PA-induced skin inflammation after treatment with EaEAC for 2 weeks. Key phenotype markers including lymph node weight, immunoglobulin E (IgE) concentration, epidermis thickness and number of infiltrated mast cells were significantly decreased in the PA+EaEAC treated group compared with the PA+Vehicle treated group. In addition, expression of IL-1ß and TNF-α was also decreased in the PA+EaEAC cotreated group, compared to PA+Vehicle treated group. Furthermore, a significant decrease in the luciferase signal derived from IL-4 promoter was detected in the abdominal region, submandibular lymph node and mesenteric lymph node of the PA+EaEAC treated group, compared to PA+Vehicle treated group. Taken together, these results suggest that EaEAC treatment could successfully improve PA-induced skin inflammation of IL-4/Luc/CNS-1 Tg mice, and that IL-4 cytokine plays a key role in the therapeutic process of EaEAC.

11.
J Ethnopharmacol ; 184: 144-53, 2016 May 26.
Artigo em Inglês | MEDLINE | ID: mdl-26969403

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Liriope platyphylla Wang et Tang continues to be used in Korea as a traditional medicine for the treatment of gastrointestinal (GI) disorders related to constipation and abnormal GI motility. AIM OF THE STUDY: Because GI disorders, especially GI motility dysfunctions, are major lifelong problems, the authors investigated the effects of a water extract of the roots of L. platyphylla Wang et Tang (LPE) on the pacemaker potentials (PPTs) of interstitial cells of Cajal (ICCs) and on GI motility in male ICR mice. MATERIALS AND METHODS: Enzymatic digestions were used to dissociate ICCs from small intestines, and the whole-cell patch-clamp configuration was used to record PPTs generated by cultured ICCs in vitro. In vivo effects of LPE on GI motility were investigated by measuring intestinal transit rates (ITRs) of Evans blue in normal mice and in acetic acid (AA) and streptozotocin (STZ)-induced diabetic mouse models of GI motility dysfunction. RESULTS: LPE dose-dependently depolarized PPTs in ICCs. Pretreatment with methoctramine (a muscarinic M2 receptor antagonist) did not block LPE-induced PPT depolarization. However, pretreatment with 4-DAMP (a muscarinic M3 receptor antagonist) blocked LPE-induced PPT depolarization. In addition, treatment with LY294002 (a phosphoinositide 3-kinase (PI3K) inhibitor) also blocked LPE-induced PPT depolarization. Intracellular GDPßS inhibited LPE-induced PPT depolarization, and LPE-induced PPT depolarization was found to occur in a phospholipase C (PLC)- and a protein kinase C (PKC)-dependent manner. Pretreatment with Ca(2+)free solution or thapsigargin (a Ca(2+)-ATPase inhibitor in endoplasmic reticulum) abolished PPTs, and under these conditions, LPE did not depolarize ICC PPTs. In normal mice, ITRs were significantly and dose-dependently increased by LPE (0.01-1g/kg administered intragastrically (i.g.)). In addition, LPE (i.g.) significantly recovered GI motility dysfunctions in both animal models. CONCLUSION: LPE dose-dependently depolarizes ICC PPTs through M3 receptors via external and internal Ca(2+)regulation and via G protein-, PI3K-, PLC- and PKC- dependent pathways in vitro. Also, in vivo, LPE increased ITRs in treatment naïve mice and our two mouse models of GI dysfunction. Therefore, this study shows that LPE offers a basis for the development of a prokinetic agent that prevents or alleviates GI motility dysfunctions.


Assuntos
Motilidade Gastrointestinal/efeitos dos fármacos , Células Intersticiais de Cajal/efeitos dos fármacos , Liriope (Planta) , Extratos Vegetais/farmacologia , Ácido Acético , Animais , Células Cultivadas , Diabetes Mellitus Experimental , Proteínas de Ligação ao GTP/fisiologia , Células Intersticiais de Cajal/fisiologia , Intestino Delgado/citologia , Intestino Delgado/fisiologia , Masculino , Camundongos Endogâmicos ICR , Fosfatidilinositol 3-Quinases/fisiologia , Raízes de Plantas , Proteína Quinase C/fisiologia , Fosfolipases Tipo C/fisiologia
12.
Lab Anim Res ; 30(2): 54-63, 2014 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-24999359

RESUMO

To investigate the toxic effects of cheonggukjang (CKJ) manufactured using mixed cultures of Bacillus subtilis MC31 and Lactobacillus sakei 383 on the liver and kidney of ICR mice, an alteration on the related markers including body weight, organ weight, urine composition, liver pathology and kidney pathology were analyzed after oral administration at dosage of 25, 50 and 100 mg/kg body weight/day of CKJ for 14 days. Any significant toxicity was not observed on the body and organ weight, clinical phenotypes, urine parameters and mortality in the CKJ-treated group compared with the vehicle-treated group. Also, liver toxicity analysis revealed no significant increase in alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartate aminotransferase (AST) or lactate dehydrogenase (LDH) in response to CKJ. Additionally, the specific pathological features induced by most toxic compounds were not observed upon liver histological analysis. Furthermore, kidney toxicological analysis revealed that blood urea nitrogen (BUN) and the serum creatinine (Cr) levels and pathological features on histological sections did not differ significantly between the vehicle- and CKJ-treated groups. Overall, these results suggest that CKJ does not induce any specific toxicity in liver and kidney organs of ICR at dose of 100 mg/kg body weight/day as no observed adverse effect level (NOAEL).

13.
Biotechnol Lett ; 36(4): 789-96, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24322767

RESUMO

A new method is presented for synthesizing arbutin glycosides using α-glucosidase (AglA) from Thermoplasma acidophilum and its glycosynthase mutant. An α-glycosynthase was constructed by substituting the catalytic nucleophile with the non-nucleophile glycine. Enzyme activity was then recovered using an external nucleophile. The transglycosylation reaction of AglA using maltose as a donor and arbutin as an acceptor produced arbutin coupled with a glucose moiety. The products were isolated and further analysed using preparative recycling HPLC. Arbutin glycosides linked to C-3, C-4, and C-6 were identified using NMR. The transglycosylation products of AglA were used as substrates for the enzyme reaction, which were hydrolyzed back again and reduced final yields. The glycosynthase mutant produced one main arbutin glycoside linked to C-4 with a yield of 38 % without further observed hydrolysis.


Assuntos
Arbutina/metabolismo , Glicoconjugados/metabolismo , Thermoplasma/enzimologia , alfa-Glucosidases/metabolismo , Biotransformação , Cromatografia Líquida de Alta Pressão , Espectroscopia de Ressonância Magnética , Maltose/metabolismo , Proteínas Mutantes/genética , Proteínas Mutantes/isolamento & purificação , Proteínas Mutantes/metabolismo , Thermoplasma/genética , alfa-Glucosidases/genética , alfa-Glucosidases/isolamento & purificação
14.
Prev Nutr Food Sci ; 19(4): 314-20, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25580396

RESUMO

Flavor quality of Liriopis tuber tea that was made using a steaming process was studied by measuring changes in headspace volatile compounds. Headspace volatile compounds of the prepared samples were isolated, separated and identified by the combined system of purge & trap, automatic thermal desorber, gas chromatography, and mass selective detector. As steaming frequencies were increased, the area percent of aldehydes decreased from 32.01% to 3.39% at 1 and 9 steaming frequency times, respectively. However, furans and ketones increased from 18.67% to 33.86% and from 9.60% to 17.40% at 1 and 9 times, respectively. The savory flavor of Liriopis tuber tea was due to a decrease in aldehydes contributing a fresh flavor at the 1st steaming process and newly generated furans from nonenzymatic browning with repeated steaming frequencies. These results will provide basic information for quality control of the newly developed Liriopis tuber tea.

15.
PLoS One ; 8(12): e78191, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24312443

RESUMO

Approximately 10-15% of individuals infected with Helicobacter pylori will develop ulcer disease (gastric or duodenal ulcer), while most people infected with H. pylori will be asymptomatic. The majority of infected individuals remain asymptomatic partly due to the inhibition of synthesis of cholesteryl α-glucosides in H. pylori cell wall by α1,4-GlcNAc-capped mucin O-glycans, which are expressed in the deeper portion of gastric mucosa. However, it has not been determined how cholesteryl α-glucosyltransferase (αCgT), which forms cholesteryl α-glucosides, functions in the pathogenesis of H. pylori infection. Here, we show that the activity of αCgT from H. pylori clinical isolates is highly correlated with the degree of gastric atrophy. We investigated the role of cholesteryl α-glucosides in various aspects of the immune response. Phagocytosis and activation of dendritic cells were observed at similar degrees in the presence of wild-type H. pylori or variants harboring mutant forms of αCgT showing a range of enzymatic activity. However, cholesteryl α-glucosides were recognized by invariant natural killer T (iNKT) cells, eliciting an immune response in vitro and in vivo. Following inoculation of H. pylori harboring highly active αCgT into iNKT cell-deficient (Jα18(-/-)) or wild-type mice, bacterial recovery significantly increased in Jα18(-/-) compared to wild-type mice. Moreover, cytokine production characteristic of Th1 and Th2 cells dramatically decreased in Jα18(-/-) compared to wild-type mice. These findings demonstrate that cholesteryl α-glucosides play critical roles in H. pylori-mediated gastric inflammation and precancerous atrophic gastritis.


Assuntos
Gastrite Atrófica/imunologia , Glucosídeos/imunologia , Infecções por Helicobacter/imunologia , Helicobacter pylori/imunologia , Helicobacter pylori/patogenicidade , Células T Matadoras Naturais/imunologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Criança , Feminino , Gastrite Atrófica/genética , Gastrite Atrófica/microbiologia , Gastrite Atrófica/patologia , Infecções por Helicobacter/genética , Infecções por Helicobacter/patologia , Helicobacter pylori/genética , Humanos , Masculino , Camundongos , Camundongos Knockout , Pessoa de Meia-Idade , Células T Matadoras Naturais/patologia , Fagocitose/imunologia , Células Th1/imunologia , Células Th1/patologia , Células Th2/imunologia , Células Th2/patologia
16.
J Microbiol Biotechnol ; 23(1): 56-63, 2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23314368

RESUMO

We have characterized the putative α-glucosidase gene (st2525) selected by total genome analysis from the acidothermophilic crenarchaeon Sulfolobus tokodaii strain 7. The ORF was cloned and expressed as a fusion protein in Escherichia coli, and recombinant ST2525 was purified by Ni-NTA affinity chromatography. Maximum activity was observed at 95°C and pH 4.0, and the enzyme exhibited stability with half-lives of 40.1 min and 7.75 min at extremely high temperatures of 100°C and 105°C, respectively. The enzyme retained at least 85% of its maximal activity in the pH range of 4.0-11.0. ST2525 exclusively hydrolyzed α-1,4-glycosidic linkages of oligosaccharides in an exo-type manner, with highest catalytic efficiency toward maltotriose. The enzyme also displayed transglycosylation activity, converting maltose to isomaltose, panose, maltotriose, isomaltotriose, etc. From these results, ST2525 could be potentially useful for starch hydrolysis as well as novel synthesis of oligosaccharides in industry.


Assuntos
Sulfolobus/enzimologia , alfa-Glucosidases/metabolismo , Cromatografia de Afinidade , Clonagem Molecular , Estabilidade Enzimática , Escherichia coli/genética , Concentração de Íons de Hidrogênio , Hidrólise , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/metabolismo , Amido/metabolismo , Especificidade por Substrato , Sulfolobus/genética , Temperatura , alfa-Glucosidases/química , alfa-Glucosidases/genética
17.
J Microbiol Biotechnol ; 22(12): 1698-704, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23221533

RESUMO

Resveratrol, or its glycoside form piceid, is a dietary antioxidant polyphenolic compound, found in grapes and red wine that has been shown to have protective effects against cardiovascular disease. However, very low water solubility of the compound may limit its application in the food and pharmaceutical industries. The amylosucrase (AMAS) of Alteromonas macleodii Deep ecotype was expressed in Escherichia coli and showed high glycosyltransferase activity to produce the glucosyl piceid when piceid was used as an acceptor. The conversion yield of piceid glucoside was 35.2%. Biotransformation using culture of the E. coli harboring the amas gene increased the yield up to 70.8%. The transfer product was purified by reverse phase chromatography and recycling preparative HPLC, and the molecular structure of the piceid glucoside was determined using NMR spectroscopy. The piceid glucoside was identified as glucosyl-alpha-(1-->4)-piceid. The solubility of glucosyl piceid was 5.26 and 1.14 times higher than those of resveratrol and piceid, respectively. It is anticipated that dietary intake of this compound is more effective by enhancing the bioavailability of resveratrol in the human body because of its hydrophilic properties in the intestinal fluid.


Assuntos
Alteromonas/enzimologia , Proteínas de Bactérias/biossíntese , Glucosídeos/química , Glucosídeos/metabolismo , Glucosiltransferases/biossíntese , Estilbenos/química , Estilbenos/metabolismo , Alteromonas/genética , Proteínas de Bactérias/genética , Proteínas de Bactérias/isolamento & purificação , Reatores Biológicos , Escherichia coli/genética , Glucosiltransferases/genética , Glucosiltransferases/isolamento & purificação , Glicosilação , Concentração de Íons de Hidrogênio , Ressonância Magnética Nuclear Biomolecular , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Solubilidade , Temperatura
18.
Lab Anim Res ; 28(3): 155-63, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23091515

RESUMO

Liriope platyphylla (LP) has long been regarded as a curative herb for the treatment of diabetes, asthma, and neurodegenerative disorders. To examine the therapeutic effects of Red LP (RLP) manufactured by steaming process on neurodegenerative disorders, significant alteration of the key factors influencing Alzheimer's Disease (AD) was detected in NSE/hAPPsw transgenic (Tg) mice after RLP treatment. The concentration of nerve growth factor (NGF) in serum increased in RLP-treated NSE/hAPPsw Tg mice compared with vehicle-treated Tg mice. However, downstream effectors of the NGF receptor signaling pathway, including TrkA and p75(NTR) proteins, were suppressed in RLP-treated NSE/hAPPsw Tg mice. Especially, Tg mice showed decreased levels of TrkA, p75(NTR), and RhoA expression. Production of Aß-42 peptides was lower in RLP-treated NSE/hAPPsw Tg mice than in vehicle-treated Tg mice. Further, analysis of γ-secretase components showed that Aß-42 peptide expression was downregulated. Of the four components, the expression of APH-1 and Nicastrin (NCT) decreased in RLP-treated NSE/hAPPsw Tg mice, whereas expression of PS-2 and Pen-2 was maintained or increased within the same group. Overall, these results suggest that RLP can help relieve neurodegenerative diseases, especially AD, through upregulation of NGF secretion ability, activation of NGF signaling pathway, downregulation of Aß-42 peptide deposition, and alteration of γ-secretase components.

19.
Lab Anim Res ; 28(3): 181-91, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23091518

RESUMO

Liriope platyphylla is a medical herb that has long been used in Korea and China to treat cough, sputum, neurodegenerative disorders, obesity, and diabetes. The aims of this study were to determine the antidiabetic and antiobesity effects of aqueous extract of L. platyphylla (AEtLP) through glucose and lipid regulation in both pre-diabetes and obesity stage of type II diabetes model. Two concentrations of AEtLP were orally administrated to OLETF (Otsuka Long-Evans Tokushima Fatty) rats once a day for 2 weeks, after which changes in glucose metabolism and fat accumulation were measured. Abdominal fat mass dramatically decreased in AEtLP-treated OLETF rats, whereas glucose concentration slightly decreased in all AEtLP-treated rats. However, compared to vehicle-treated OLETF rats, only AEtLP10 (10% concentration)-treated OLETF rats displayed significant induction of insulin production, whereas AEtLP5 (5% concentration)-treated OLETF rats showed a lower level of insulin. Although serum adiponectin level increased in only AEtLP5-treated rats, significant alteration of lipid concentration was detected in AEtLP5-treated OLETF rats. Expression of Glut-1 decreased in all AEtLP-treated rats, whereas Akt phosphorylation increased only in AEtLP10-treated OLETF rats. Furthermore, the pattern of Glut-3 expression was very similar with that of Glut-1 expression, which roughly corresponded with the phosphorylation of c-Jun N-teminal kinase (JNK) and p38 in the mitogen-activated protein kinase pathway. Therefore, these findings suggest that AEtLP should be considered as a therapeutic candidate during pre-diabetes and obesity stage capable of inducing insulin secretion from pancreatic ß-cells, glucose uptake in liver cells, as well as a decrease in fat and lipid accumulation.

20.
Lab Anim Res ; 28(4): 229-38, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23326283

RESUMO

Red Liriope platyphylla (RLP) produced by steaming process has been reported to enhance the secretion of insulin and nerve growth factor (NGF). However, there has been no report on the toxicity of RLP in the specific organs of mice. To investigate the toxic effect of RLP, we tried to observe a significant alteration on body weight, food/water intake, organ weight, liver pathology and kidney pathology in female ICR mice received 12.5, 25.0 and 50.0 mg/kg body weight/day of RLP via gavage for 10 days. Out of seven organs including brain, heart, lung, liver, kidney, spleen and ovary, two organs (heart and lung) showed significantly decreased weights in the medium dosage RLP-treated group, whereas weights of other organs were maintained at constant levels in all dosage groups. In the liver toxicity analysis, no significant increase of alkaline phosphatase (ALP), alanine aminotransferase (ALT), and aspartate amino-transferase (AST) were detected in any RLP-treated group compared to vehicle-treated group. The specific pathological changes induced by most of toxic compounds were not observed in the liver in microscopic examination. Furthermore, in the kidney toxicological analysis, a significant enhancement of the blood urea nitrogen (BUN) concentration was detected in the high dosage RLP-treated group compared to the vehicle-treated group. However, the serum creatinine (CA) concentration on the serum biochemistry as well as the pathological changes in microscopic examination were not significantly different between the vehicle- and RLP-treated groups. Therefore, these results suggest that RLP does not induce any specific toxicity in liver or kidney tissues of mice, although the BUN level slightly increased in 50.0 mg/kg of RLP-treated group.

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