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1.
Nutrients ; 16(5)2024 Mar 05.
Article in English | MEDLINE | ID: mdl-38474877

ABSTRACT

In this study, the roles of kiwifruit soluble/insoluble dietary fiber (SDF/IDF, respectively) in the pasting characteristics and in vitro digestibility of wheat starch were explored. According to RVA and rheological tests, the IDF enhanced the wheat starch viscosity, decreased the gelatinization degree of the starch granules, and exacerbated starch retrogradation. The addition of SDF in high quantities could reduce the starch gelatinization level, lower the system viscosity, and exacerbate starch retrogradation. Through determining the leached amylose content and conducing scanning electron microscopy, the IDF and SDF added in high quantities was combined with the leached amylose wrapped around the starch granules, which reduced the leached amylose content and decreased the gelatinization degree of the starch granules. The Fourier transform infrared results showed that the addition of both the IDF and SDF resulted in an enhancement in hydrogen bonding formed by the hydroxyl groups of the system. The in vitro digestion results strongly suggested that both the IDF and SDF reduced the wheat starch digestibility. The above findings are instructive for the application of both IDF and SDF in starchy functional foods.


Subject(s)
Amylose , Starch , Triticum , Dietary Fiber , Viscosity
2.
J Sci Food Agric ; 104(6): 3246-3255, 2024 Apr.
Article in English | MEDLINE | ID: mdl-38081762

ABSTRACT

BACKGROUND: The aim of this research was to evaluate the possibility of lipid concomitant γ-oryzanol reducing oil absorbency of fried foods and the underlying mechanism. Therefore, the influence of γ-oryzanol on moisture and oil content, and distribution and micromorphology of French fries and the viscosity, fatty acid composition and total polar compounds content of rice bran oil (RBO) after frying were studied. RESULTS: Our results showed that the incorporation of low concentration of γ-oryzanol [low addition group (LAG)] (5.754 g/kg) decreased the oil absorbency and porous structure of French fries during frying. Additionally, LAG incorporation inhibited the degradation of linoleic acid, decreased the growth rate of saturated fatty acids, total polar compounds and viscosity of frying oil. CONCLUSIONS: Consequently, it was recommended to incorporate a small amount of γ-oryzanol in frying oil because it could inhibit oil absorption behavior of French fries. © 2023 Society of Chemical Industry.


Subject(s)
Cooking , Phenylpropionates , Cooking/methods , Fatty Acids , Rice Bran Oil
3.
Microb Pathog ; 184: 106323, 2023 Nov.
Article in English | MEDLINE | ID: mdl-37633505

ABSTRACT

Salmonella enterica serovar Typhimurium (S. typhimurium) is a common Gram-negative foodborne pathogen that threatens public health and hinders the development of livestock industry. Resveratrol, an important component in grape fruits and seeds, has been shown to possess multiple biological activities, but its potential effects on S. typhimurium-mediated virulence have been rarely reported. In this study, we investigated the effect of resveratrol on S. typhimurium flagella -mediated virulence. The results showed that resveratrol significantly reduced the transcription of flagella genes and swimming motility of S. typhimurium, and also inhibited the transcription of T3SS-related virulence genes with varying degrees inhibiting bacterial growth. Simultaneously, resveratrol significantly reduced the adhesion of S. typhimurium to HeLa cells. Unfortunately, resveratrol does not improve the survival rate of S. typhimurium-infected mice, but it reduces the bacterial load in the liver and spleen of infected mice, and it also has a certain degree of anti-inflammatory activity. In summary, these results indicated that resveratrol has the potential to be developed as an alternative drug or antibacterial agent to prevent Salmonella infection.


Subject(s)
Salmonella Infections, Animal , Swimming , Humans , Animals , Mice , Resveratrol/pharmacology , HeLa Cells , Serogroup , Salmonella typhimurium/genetics , Salmonella Infections, Animal/drug therapy , Salmonella Infections, Animal/microbiology , Bacterial Proteins/genetics
4.
Front Aging Neurosci ; 15: 1139418, 2023.
Article in English | MEDLINE | ID: mdl-37113572

ABSTRACT

Introduction: The misfolding and aggregation of ß-amyloid (Aß) easily form Aß fibers, which are continuously deposited in the brain, leading to the massive generation of amyloid plaques, severely destroying neuronal connections, and promoting Alzheimer's disease (AD) The occurrence and development of AD is one of the pathogenesis of AD. There is an urgent need to develop inhibitors against Aß aggregation, which is hopefully a potential way to treat AD. Methods: In this study, we first found the crystal structure of the Aß1-42 receptor protein from the RCSB PDB protein structure database and used the SYBYL X2.0 software for molecular docking, and then used the Peptide Ranker, Innovagen, DPL, and ToxinPred online websites to perform peptides. Predict the activity score, toxicity and water solubility, and then calculate the affinity constant KD value of polypeptide and Aß through Surface Plasmon Resonance (SPR) experiment. Subsequently, the CCK-8 kit method was used to determine the toxicity of different concentrations of peptides (3.125, 6.25, 12.5, 25, 50, 100, 200 µM) to PC12 cells, and then the peptides and Aß according to different concentration ratios (1:4, 1:2, 1:1, 1:0.5, 1:0.25, 0:4), this method is also used to detect the effect of peptides on Aß-induced neurotoxicity. The thioflavin T (ThT) fluorescence method was used to detect the effects of peptides (50 µM) on Aß (25 µM) aggregation inhibitory effect. Results: The results showed that the CScore of YVRHLKYVRHLK peptide molecule docking was 10.0608, the predicted activity score was 0.20, and the KD value was 5.385 × 10-5. The ThT and CCK-8 kit method found that the peptide itself is less toxic to PC12 cells at a concentration of 50 µM, and it has a significant inhibitory effect on the formation of Aß1-42 aggregates when incubated with Aß1-42 at a ratio of 1:1 (p < 0.05) and can significantly reduce the PC12 cytotoxicity induced by Aß1-42 (p < 0.05). Conclusion: In conclusion, the polypeptide YVRHLKYVRHLK designed in this study has a neuroprotective effect on PC12 cytotoxicity induced by Aß1-42. Graphical Abstract.

5.
Nutrients ; 15(4)2023 Feb 06.
Article in English | MEDLINE | ID: mdl-36839182

ABSTRACT

The purposes of the present work are to assess how 6-gingerol (6G) positively influences serum glucose regulation in mice with prediabetes triggered by streptozotocin (STZ) plus a high-fat diet (HFD) and to clarify its underlying mechanisms. An analysis of prediabetic symptoms and biochemical characteristics found that 6G intervention was significantly associated with reduced fasting glucose levels, alleviated insulin resistance, better glucose tolerance, hepatic and pancreatic impairment, and dyslipidemia. For the recognition of the target gut microbiota and the pathways linked to 6G's hypoglycemic function, a combination of hepatic RNA and 16S rRNA sequencing was employed. Specifically, 6G significantly improved the dysbiosis of the gut microbiota and elevated the relative abundances of Alistipes, Alloprevotella, and Ruminococcus_1. Furthermore, 6G supplementation inhibited gluconeogenesis and stimulated glycolysis by activating the PI3K/AKT axis, which also repressed the oxidative stress through Nrf2/Keap1-axis initiation. In addition, Spearman's correlation analyses reveal a complex interdependency set among the gut microbiota, metabolic variables, and signaling axes. Taken together, the hypoglycemic effect of 6G is partially mediated by altered gut microbiota, as well as by activated Nrf2/Keap1 and PI3K/AKT axes. Thus, 6G may be used as a candidate dietary supplement for relieving prediabetes.


Subject(s)
Diet, High-Fat , Prediabetic State , Mice , Animals , Streptozocin , Kelch-Like ECH-Associated Protein 1 , Proto-Oncogene Proteins c-akt , RNA, Ribosomal, 16S/genetics , NF-E2-Related Factor 2 , Phosphatidylinositol 3-Kinases , Bacteroidetes/genetics , Glucose , Mice, Inbred C57BL
6.
Nutrients ; 14(16)2022 Aug 17.
Article in English | MEDLINE | ID: mdl-36014875

ABSTRACT

This study aims to examine the anti-diabetic properties of insoluble and soluble dietary fibers from kiwifruit (KIDF and KSDF) in rats with type 2 diabetes mellitus (T2DM) resulting from a high-fat diet (HFD) and streptozotocin (STZ). Both KIDF and KSDF treatments for four weeks remarkably decreased body weight and increased satiety. In addition, the blood glucose level and circulatory lipopolysaccharide (LPS) content were decreased, while the insulin resistance, inflammatory status, and lipid profiles improved. These anti-diabetic effects might be related to the regulation of gut microbiota and increased SCFA content. The key microbial communities of KIDF and KSDF were different. Furthermore, the KIDF treatment increased the level of total SCFAs and isobutyric acid, while KSDF increased the levels of total SCFAs and butyric acid. The association between critical species and SCFA and between SCFA and biochemical parameters indicated that the mechanisms of KIDF and KSDF on T2DM might be different.


Subject(s)
Actinidia , Diabetes Mellitus, Type 2 , Gastrointestinal Microbiome , Animals , Butyric Acid/pharmacology , Diet, High-Fat/adverse effects , Dietary Fiber/pharmacology , Rats , Streptozocin
7.
Front Pharmacol ; 13: 862618, 2022.
Article in English | MEDLINE | ID: mdl-35677434

ABSTRACT

Osteoblast dysfunction, induced by high glucose (HG), negatively impacts bone homeostasis and contributes to the pathology of diabetic osteoporosis (DOP). One of the most widely recognized mechanisms for osteoblast dysfunction is oxidative stress. Resveratrol (RES) is a bioactive antioxidant compound to combat oxidative damage. However, its role in the protection of HG-induced osteoblast dysfunction has not been clarified. Therefore, our study aimed to explore potential regulatory mechanisms of RES for attenuating HG-induced osteoblast dysfunction. Our results showed that osteoblast dysfunction under HG condition was significantly ameliorated by RES via the activation of nuclear factor erythroid 2-related factor (NRF2) to suppress oxidative stress. Furthermore, using Nrf2-shRNA and wortmannin, we identified that activation of NRF2 via RES was regulated by the AKT/glycogen synthase kinase 3ß (GSK3ß)/FYN axis.

8.
Food Chem ; 387: 132914, 2022 Sep 01.
Article in English | MEDLINE | ID: mdl-35421650

ABSTRACT

Bovine ß-casein (ß-CN) has attracted increasingly interest as biocompatible nanocarrier for hydrophobic flavonoid due to its self-assembly ability to form micelles. This paper reported Maillard induced glycation reaction of ß-CN using dextran in order to improve stability of naringenin-loaded ß-CN micelles under acidic and high calcium environments. Our results showed that solubility of ß-CN-graft-dextran was remarkable increased at acidic pH and the conjugation with 20 kDa dextran had the highest level of graft degree. Glycation restrained ß-CN from aggregating around pH 5.0 where was close to the isoelectric point, forming spherical micelles with irregular and rough surfaces, which were significantly larger than the micelles at pH 7.0. ß-CN-graft-dextran also overcame destabilization of the micelles induced by excess calcium and had no impact on the chelating ability of calcium. These findings appeared to be promising for future applications of modified ß-CN-graft-dextran based on Maillard reaction as fairly stable nanocarrier under extreme condition.


Subject(s)
Caseins , Micelles , Animals , Calcium/chemistry , Calcium, Dietary , Caseins/chemistry , Cattle , Dextrans/chemistry , Maillard Reaction
9.
J Food Sci ; 87(3): 1268-1281, 2022 Mar.
Article in English | MEDLINE | ID: mdl-35152443

ABSTRACT

The present study aimed to examine the protective effect of Bentong ginger oleoresin (BGO) on the occurrence of nonalcoholic fatty liver disease (NAFLD) and its underlying mechanism. In the present study, 14-week BGO treatment reduced the high-fat diet (HFD)-induced obesity. The serum total cholesterol (TC) was reduced from 4.76 ± 0.30 to 3.542 ± 0.49 mmol/L and fatty liver score decreased to the normal level (1.6 ± 0.55). BGO had antihypercholesterolemia activity, alleviated abnormal lipid metabolism, and improved liver fat accumulation. In addition, liver inflammatory cytokine tests and Western blotting analysis indicated that BGO might play an anti-inflammatory role by mediating the NF-κB signaling pathway. Moreover, BGO regulated the gut microbiota in NAFLD mice and finally mediated their benefits for the host, which might be associated with reduced abundance of Lachnospiraceae_NK4A136_group and Fournierella. BGO showed effective liver protection and regulation of gut microbiota for the HFD-induced NAFLD in obese mice. As a result, BGO may serve as an effective dietary supplement for the improvement of NAFLD-related metabolic diseases. PRACTICAL APPLICATION: This study provides a new way to improve the added value of Bentong ginger. It also provides certain experimental data on BGO as a kind of the functional food ingredient. The current work also provides new ideas for the improvement and treatment of NAFLD.


Subject(s)
Gastrointestinal Microbiome , Non-alcoholic Fatty Liver Disease , Zingiber officinale , Animals , Diet, High-Fat/adverse effects , Liver/metabolism , Mice , Mice, Inbred C57BL , Non-alcoholic Fatty Liver Disease/drug therapy , Non-alcoholic Fatty Liver Disease/metabolism , Plant Extracts
10.
Food Chem ; 326: 127028, 2020 Oct 01.
Article in English | MEDLINE | ID: mdl-32428857

ABSTRACT

One interesting phenomenon of Chrysanthemum morifolium tea is its formation of a green or dark green color after hours of brewing, and it is important to study the compounds that form this color and whether they might be involved in health benefits. Non-targeted metabolomics could clearly distinguish between green and non-green species by sparse partial least squares discriminant analysis (sPLS-DA) and heat-mapping. We found that the pH was the primary factor in the formation of the green color. Two green precursors (GP1 and GP2) were separated and purified with preparative HPLC. FT-IR analysis and the saccharide content analysis showed that GP1 had the typical characteristics of saccharides. GP2 was identified as chlorogenic acid by ESI-Q-TOF/MS and NMR. We raised the formation process of green substances was caused by the hydrolysates of chlorogenic acid reacting with glycosides or groups attached to the saccharides, which suggests a new mechanism for color-forming reactions.


Subject(s)
Chrysanthemum/chemistry , Tea/chemistry , Chromatography, High Pressure Liquid , Color , Flowers/chemistry , Glycosides/analysis , Least-Squares Analysis , Plant Extracts/chemistry , Spectroscopy, Fourier Transform Infrared
11.
Planta ; 251(5): 95, 2020 Apr 09.
Article in English | MEDLINE | ID: mdl-32274590

ABSTRACT

MAIN CONCLUSION: Formation of specific ultrastructural chromoplastidal elements during ripening of fruits of three different colored Physalis spp. is closely related to their distinct carotenoid profiles. The accumulation of color-determining carotenoids within the chromoplasts of ripening yellow, orange, and red fruit of Physalis pubescens L., Physalis peruviana L., and Physalis alkekengi L., respectively, was monitored by high-performance liquid chromatography/diode array detector/tandem mass spectrometry (HPLC-DAD-MS/MS) as well as light and transmission electron microscopy. Both yellow and orange fruit gradually accumulated mainly ß-carotene and lutein esters at variable levels, explaining their different colors at full ripeness. Upon commencing ß-carotene biosynthesis, large crystals appeared in their chromoplasts, while large filaments protruding from plastoglobules were characteristic elements of chromoplasts of orange fruit. In contrast to yellow and orange fruit, fully ripe red fruit contained almost no ß-carotene, but esters of both ß-cryptoxanthin and zeaxanthin at very high levels. Tubule bundles and unusual disc-like crystallites were predominant carotenoid-bearing elements in red fruit. Our study supports the earlier hypothesis that the predominant carotenoid type might shape the ultrastructural carotenoid deposition form, which is considered important for color, stability and bioavailability of the contained carotenoids.


Subject(s)
Carotenoids/analysis , Fruit/growth & development , Physalis/growth & development , Chromatography, High Pressure Liquid , Color , Fruit/physiology , Fruit/ultrastructure , Lutein/analysis , Physalis/physiology , Physalis/ultrastructure , Pigmentation , Plastids/ultrastructure , Tandem Mass Spectrometry , Zeaxanthins/analysis , beta Carotene/analysis
12.
Chemosphere ; 244: 125495, 2020 Apr.
Article in English | MEDLINE | ID: mdl-31837563

ABSTRACT

Dysregulated activation of inflammation plays an important role in the development and progression of neuronal damage, and limiting the production of reactive oxygen species (ROS) can suppress the inflammatory signals. Nuclear factor erythroid 2-related factor 2 (Nrf2) is a redox-sensing transcription factor that drives an adaptive cellular defense in response to oxidative stress. However, the implications of Nrf2 in sodium fluoride (NaF)-stimulated microglia and the underlying mechanisms remain obscure. In this study, we demonstrated that NaF activated the Nrf2 signaling and enhanced the downstream antioxidant protein levels, including heme oxygenase-1 and quinine oxidoreductase 1. NaF induced oxidative stress, as indicated by increased ROS level and malondialdehyde content, and reduced superoxide dismutase activity. Moreover, NaF promoted the nuclear translocation of NF-κB, thus increased the production of the pro-inflammatory cytokines tumor necrosis factor-α, interleukin (IL)-6, and IL-1ß. However, these effects were relieved by overexpression of Nrf2. Meanwhile, knockdown of Nrf2 by shRNA exacerbated NaF-induced oxidative stress and inflammation in BV-2 cells and primary cultured microglia. Mechanistically, NaF-induced Nrf2 activation is AMPK/p38 dependent, as deletion of AMPK using siRNA blocked the activating effect of NaF on p38 and Nrf2. Notably, treatment of N-Acety-l-Cysteine attenuated AMPK/p38-dependent Nrf2 activation in microglia exposed to NaF. In conclusion, these data demonstrated for the first time that Nrf2 activation exerts a neuroprotective effect on NaF-stimulated redox imbalance and inflammation that is dependent on the AMPK/p38 pathway.


Subject(s)
AMP-Activated Protein Kinases/metabolism , Microglia/physiology , NF-E2-Related Factor 2/metabolism , Oxidative Stress/physiology , Sodium Fluoride/toxicity , Animals , Antioxidants/metabolism , Cytokines/metabolism , Heme Oxygenase-1/genetics , Inflammation/metabolism , Interleukin-6/metabolism , Lipopolysaccharides/metabolism , NF-kappa B/metabolism , Neuroprotective Agents/metabolism , Oxidative Stress/drug effects , Reactive Oxygen Species/metabolism , Signal Transduction/drug effects , Tumor Necrosis Factor-alpha/metabolism
13.
Food Chem ; 289: 657-663, 2019 Aug 15.
Article in English | MEDLINE | ID: mdl-30955661

ABSTRACT

Polyphenol oxidase from Granny Smith apples was purified and characterized in both its soluble form (sPPO) and its membrane-bound form (mPPO). Both forms were purified by temperature-induced phase partitioning, precipitation with ammonium sulfate, and ion exchange chromatography. The specific activity of mPPO was 19.17 times that of sPPO. The optimum pH and temperature for both forms were 7.0 and 35 °C when catechol was the substrate. The Michaelis constant and maximum reaction rate for sPPO were 34.1 mM and 500 U/mL/min, whereas those for mPPO were 53 mM and 10,000 U/mL/min, respectively. The enzymes exhibited diphenolase activity, and their affinity was highest for catechol (sPPO) and 4-methylcatechol (mPPO). Inhibitors of sPPO and mPPO included ascorbic acid, glutathione, and l-cysteine. However, ethylenediaminetetraacetic acid increased the activity of mPPO. Purified sPPO was dimeric with a molecular weight of 31 kDa, whereas mPPO was monomeric with an estimated molecular weight of 65 kDa.


Subject(s)
Catechol Oxidase/metabolism , Fruit/enzymology , Malus/enzymology , Ascorbic Acid/metabolism , Catechols/chemistry , Cysteine/metabolism , Edetic Acid/metabolism , Glutathione/metabolism , Hydrogen-Ion Concentration , Molecular Weight , Plant Proteins/metabolism , Substrate Specificity , Temperature
14.
Food Res Int ; 120: 389-398, 2019 06.
Article in English | MEDLINE | ID: mdl-31000254

ABSTRACT

Physicochemical characteristics and phytochemical profiles of red (Physalis alkekengi L., RP) and yellow (P. pubescens L., YP) Physalis fruits cultivated in three provinces of China were characterized. YP fruits showed significantly lower levels of total organic acids and elevated total sugars than those of RP. A total of 18 compounds was tentatively identified in hydromethanolic extracts of Physalis fruits applying HPLC-DAD-ESI-MSn and HR-ESI-MS. Cinnamoyl and hydroxycinnamoyl conjugates prevailed in both fruits. Diverse mono- and dihexosides of cinnamic, coumaric, caffeic, ferulic, and sinapic acid were found in YP, while RP merely contained feruloyl and sinapoyl hexosides. N,N´-dicaffeoylspermidine isomers were found in YP fruits, whereas N,N´-bis(dihydrocaffeoyl)spermine was exclusively detected in two of the RP samples. Additionally, two HDMF (4-hydroxy-2,5-dimethyl-3(2H)-furanone) hexosides were tentatively identified for the first time in YP. Both RP and YP fruits collected from three different provinces in China showed a significant intraspecific variability regarding their phytochemical profiles, despite their similar morphological fruit traits.


Subject(s)
Fruit/chemistry , Physalis/chemistry , Phytochemicals/analysis , Amino Acids/analysis , Ascorbic Acid/analysis , China , Chromatography, High Pressure Liquid , Citric Acid/analysis , Color , Consumer Behavior , Coumaric Acids/analysis , Food Analysis , Food Quality , Fructose/analysis , Glucose/analysis , Hardness , Hydrogen-Ion Concentration , Malates/analysis , Mass Spectrometry , Quinic Acid/analysis , Sucrose/analysis , Tartrates/analysis , Taste
15.
Biomed Pharmacother ; 111: 527-536, 2019 Mar.
Article in English | MEDLINE | ID: mdl-30597306

ABSTRACT

Acute liver injury (ALI) is a life-threatening syndrome accompanied by overwhelming inflammation. Amygdalin (AGD) has been reported to possess various biological activities, particularly anti-inflammatory activity. The current study was designed to assess the protective effects and underlying mechanisms of AGD against ALI induced by d-galactosamine (GalN) and lipopolysaccharide (LPS) in mice. The results indicated that AGD treatment effectively reduced the lethality, ameliorated the histopathological liver changes, reduced the malondialdehyde (MDA) and myeloperoxidase (MPO) levels, and decreased the alanine transaminase (ALT) and aspartate aminotransferase (AST) levels resulting from LPS/GalN challenge. Moreover, AGD significantly inhibited LPS/GalN-induced inflammatory responses in mice with ALI by reducing not only the secretion of tumour necrosis factor (TNF)-α, interleukin (IL)-1ß, and IL-6 but also the protein expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). Additionally, our results demonstrated that the inhibitory effect of AGD was due to the suppressed activation of nuclear factor-kappa B (NF-κB) and nucleotide-binding domain (NOD-)like receptor protein 3 (NLRP3) inflammasome activity. Furthermore, AGD treatment substantially increased nuclear factor erythroid 2-related factor 2 (Nrf2) nuclear translocation and enhanced NAD (P) H: quinoneoxidoreductase 1 protein expression, which was reversed by a Nrf2 inhibitor, in HepG2 cells. In summary, our investigations suggested that the ability of AGD to ameliorate LPS/GalN-induced ALI may involve the inhibition of the NLRP3 inflammasome and NF-κB signalling pathways and the upregulation of the Nrf2/NQO1 signalling pathway.


Subject(s)
Amygdalin/therapeutic use , Chemical and Drug Induced Liver Injury/metabolism , Galactosamine/toxicity , NF-E2-Related Factor 2/physiology , NF-kappa B/physiology , NLR Family, Pyrin Domain-Containing 3 Protein/physiology , Amygdalin/pharmacology , Animals , Antineoplastic Agents, Phytogenic/toxicity , Chemical and Drug Induced Liver Injury/prevention & control , Dose-Response Relationship, Drug , Female , Hep G2 Cells , Humans , Lipopolysaccharides/toxicity , Mice , Mice, Inbred BALB C , NAD(P)H Dehydrogenase (Quinone)/physiology , NF-kappa B/antagonists & inhibitors , NLR Family, Pyrin Domain-Containing 3 Protein/antagonists & inhibitors , Signal Transduction/drug effects , Signal Transduction/physiology
16.
Int J Biol Macromol ; 114: 1056-1063, 2018 Jul 15.
Article in English | MEDLINE | ID: mdl-29625219

ABSTRACT

This study determined the optimal conditions for ultrasound-assisted extraction of a water-soluble polysaccharide, Raw Okra Polysaccharide, from the fruit of okra using response surface methodology. The optimal extraction temperature, extraction time and ultrasonic power were 59°C, 30min and 522W, respectively, giving a yield of 10.35±0.11%. ROP was further isolated, lyophilized and purified using a DEAE-Sepharose Fast Flow column and Sepharose CL-6B column, revealing three elution peaks subsequently designated ROP -1, -2, and -3, respectively. Of these, ROP-2 showed the highest yield, and was therefore selected for physicochemical analysis and evaluation of antioxidant activity. Gas chromatography, fourier transform infrared spectroscopy, and high-performance liquid chromatography were used to characterize the primary structural features and molecular weight, revealing that ROP-2 is composed of glucose, mannose, galactose, arabinose, xylose, fructose, and rhamnose (molar percentages: 28.8, 12.5, 13.1, 15.9, 9.2, 13.7, and 6.8%, respectively) and has an average molecular weight of 1.92×105Da. A superoxide radical scavenging assay and DPPH radical scavenging assay further revealed the significant in vitro antioxidant activity of ROP-2. These findings present an effective technique for extraction of the natural antioxidant ROP-2, warranting further analysis of its potential application in the food industry.


Subject(s)
Abelmoschus/chemistry , Antioxidants , Polysaccharides , Ultrasonic Waves , Antioxidants/chemistry , Antioxidants/isolation & purification , Polysaccharides/chemistry , Polysaccharides/isolation & purification
17.
J Pharmacol Sci ; 136(4): 203-211, 2018 Apr.
Article in English | MEDLINE | ID: mdl-29551284

ABSTRACT

Acute lung injury (ALI) arises from uncontrolled pulmonary inflammation with high mortality rates. Atractylodin (Atr) is a polyethylene alkynes and has been reported to possess anti-inflammation effect. Thus, we aimed to investigate the protective effect of Atr on lipopolysaccharide (LPS)-induced inflammatory responses ALI. The results indicated that Atr treatment not only significantly attenuated LPS-stimulated histopathological changes but also lessened the myeloperoxidase (MPO) activity, the wet-to-dry weight ratio of the lungs, protein leakage and infiltration of inflammatory cells. Moreover, Atr inhibited the tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-1ß and monocyte chemoattractant protein (MCP)-1 secretion in BALF. Further study demonstrated that such inhibitory effects of Atr were due to suppression of nucleotide-binding domain-(NOD-) like receptor protein 3 (NLRP3) inflammasome and toll like receptor 4 (TLR4) activation, likely contributing to its anti-inflammatory effects. Collectively, these findings suggest that Atr may be an effective candidate for alleviating LPS-induced inflammatory responses.


Subject(s)
Acute Lung Injury/chemically induced , Acute Lung Injury/drug therapy , Furans/pharmacology , Furans/therapeutic use , Lipopolysaccharides/adverse effects , Signal Transduction/drug effects , Acute Lung Injury/genetics , Acute Lung Injury/pathology , Animals , Anti-Inflammatory Agents , Bronchoalveolar Lavage Fluid , Cytokines/metabolism , Disease Models, Animal , Inflammasomes , Male , Mice, Inbred BALB C , NLR Family, Pyrin Domain-Containing 3 Protein , Peroxidase/metabolism , Phytotherapy , Toll-Like Receptor 4
19.
PLoS One ; 11(5): e0156260, 2016.
Article in English | MEDLINE | ID: mdl-27228340

ABSTRACT

Chronic myeloid leukemia (CML) is a myeloproliferative disease. Imatinib (IM), the first line treatment for CML, is excessively expensive and induces various side effects in CML patients. Therefore, it is essential to investigate a new strategy for improving CML therapy. Our immunoblot data revealed that RanGTPase activating protein 1 (RanGAP1) protein levels increased by approximately 30-fold in K562 cells compared with those in normal cells. RanGAP1 is one of the important components of RanGTPase system, which regulates the export of nuclear protein. However, whether RanGAP1 level variation influences BCR-ABL nuclear export is still unknown. In this report, using shRNA to downregulate RanGAP1 expression level augmented K562 cell apoptosis by approximately 40% after treatment with 250 nM IM. Immunofluorescence assay also indicated that three-fold of nuclear BCR-ABL was detected. These data suggest that BCR-ABL nuclear entrapment induced by RanGAP1 downregulation can be used to improve IM efficacy. Moreover, our qRT-PCR data indicated a trend of inverse correlation between the RanGAP1 and microRNA (miR)-1301 levels in CML patients. MiR-1301, targeting the RanGAP1 3' untranslated region, decreased by approximately 100-fold in K562 cells compared with that in normal cells. RanGAP1 downregulation by miR-1301 transfection impairs BCR-ABL nuclear export to increase approximately 60% of cell death after treatment of 250 nM IM. This result was almost the same as treatment with 1000 nM IM alone. Furthermore, immunofluorescence assay demonstrated that Tyr-99 of nuclear P73 was phosphorylated accompanied with nuclear entrapment of BCR-ABL after transfection with RanGAP1 shRNA or miR-1301 in IM-treated K562 cells. Altogether, we demonstrated that RanGAP1 downregulation can mediate BCR-ABL nuclear entrapment to activate P73-dependent apoptosis pathway which is a novel strategy for improving current IM treatment for CML.


Subject(s)
Cell Nucleus/metabolism , Drug Resistance, Neoplasm/genetics , Fusion Proteins, bcr-abl/metabolism , GTPase-Activating Proteins/metabolism , Imatinib Mesylate/pharmacology , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/pathology , MicroRNAs/genetics , Antineoplastic Agents/pharmacology , Apoptosis/drug effects , Cell Nucleus/genetics , Cell Proliferation/drug effects , Fusion Proteins, bcr-abl/genetics , GTPase-Activating Proteins/genetics , Gene Expression Regulation, Neoplastic/drug effects , Humans , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/drug therapy , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/genetics , Phosphorylation , RNA, Small Interfering/genetics , Tumor Cells, Cultured
20.
Biosens Bioelectron ; 26(8): 3511-6, 2011 Apr 15.
Article in English | MEDLINE | ID: mdl-21398106

ABSTRACT

Nitroreductase (NTR) is a flavin-containing enzyme that uses NADH as the electron source to reduce nitroaromatic compounds to the corresponding amines. Previous studies have shown that nitroreductase-targeted latent fluorophores exhibit low solubility in the aqueous media and fluoresce at lower wavelengths upon uncloaking, thus limiting their effective applications. Here, we have prepared a new switch-on long-wavelength latent fluorogenic substrate, NTRLF (4), for NTR. In the presence of NADH, NTR catalyzes the reduction of the nitroaromatic moiety in NTRLF (4), followed by the cascade reaction, 1,6-rearrangement-elimination reaction, cyclic urea formation, and concomitant ejects a long-wavelength fluorescence coumarin (8). However, this reaction was inhibited in the presence of nitroaromatic analogues. The fluorescence signal generated by the cascade reaction was specific and insensitive to various reductants. Accordingly, we propose that NTRLF and NTR in the presences of NADH constitute a useful switch-off high-throughput fluorescence sensor for screening nitroaromatic compounds. Furthermore, NTRLF in the NTR-coupled 3-hydroxybutyrate dehydrogenase and aldehyde dehydrogenase assay reactions was a sensitive fluorimetric indicator for the quantitatively measurement of 3-hydroxybutyrate and propionaldehyde, respectively within micromolar range. Our novel NTRLF and NTR-coupled dehydrogenase assay platform may thus be effectively applied for the quantitative estimation of a broad range of analytes.


Subject(s)
3-Hydroxybutyric Acid/analysis , Aldehydes/analysis , Biosensing Techniques/methods , Fluorescent Dyes , Fluorometry/methods , Nitroreductases/chemistry , Aldehyde Dehydrogenase/chemistry , Fluorescence , Hydroxybutyrate Dehydrogenase/chemistry , NAD/chemistry , Nitro Compounds/analysis
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