AIM:To investigate the effects of dihydromyricetin(DHM)on the migration and invasion of human hepatic cancer cells.METHODS:The hepatoma cell lines SK-Hep-1 and MHCC97L were used in this study.The cells were cultured i...AIM:To investigate the effects of dihydromyricetin(DHM)on the migration and invasion of human hepatic cancer cells.METHODS:The hepatoma cell lines SK-Hep-1 and MHCC97L were used in this study.The cells were cultured in RPIM-1640 medium supplemented with 10%fetal bovine serum at 37℃in a humidified 5%CO2incubator.DHM was dissolved in dimethyl sulfoxide and diluted to various concentrations in medium before applying to cells.MTT assays were performed to measure the viability of the cells after DHM treatment.Wound healing and Boyden transwell assays were used to assess cancer cell motility.The invasive capacity of cancer cells was measured using Matrigel-coated transwell chambers.Matrix metalloproteinase(MMP)-2/9 activity was examined by fluorescence analysis.Western blot was carried out to analyze the expression of MMP-2,MMP-9,p-38,JNK,ERK1/2 and PKC-δproteins.All data were analyzed by Student’s t tests in GraphPad prism 5.0software and are presented as mean±SD.RESULTS:DHM was found to strongly inhibit the migration of the hepatoma cell lines SK-Hep-1(without DHM,24 h:120±8μmol/L vs 100μmol/L DHM,24h:65±10μmol/L,P<0.001)and MHCC97L(without DHM,24 h:126±7μmol/L vs 100μmol/L DHM,24h:74±6μmol/L,P<0.001).The invasive capacity of the cells was reduced by DHM treatment(SK-Hep-1cells without DHM,24 h:67±4μmol/L vs 100μmol/L DHM,24 h:9±3μmol/L,P<0.001;MHCC97L cells without DHM,24 h:117±8μmol/L vs 100μmol/L DHM,24 h:45±2μmol/L,P<0.001).MMP2/9 activity was also inhibited by DHM exposure(SK-Hep-1 cells without DHM,24 h:600±26μmol/L vs 100μmol/L DHM,24 h:100±6μmol/L,P<0.001;MHCC97L cells without DHM,24 h:504±32μmol/L vs 100μmol/L DHM 24 h:156±10μmol/L,P<0.001).Western blot analysis showed that DHM decreased the expression level of MMP-9 but had little effect on MMP-2.Further investigation indicated that DHM markedly reduced the phosphorylation levels of p38,ERK1/2 and JNK in a concentration-dependent manner but had no impact on the total protein levels.In addition,PKC-δprotein,a key protein in the regulation of MMP family protein expression,was up-regulated with DHM treatment.CONCLUSION:These findings demonstrate that DHM inhibits the migration and invasion of hepatoma cells and may serve as a potential candidate agent for the prevention of HCC metastasis.展开更多
AIM: To assess the effects of dihydromyricetin(DHM) as a hepatoprotective candidate in reducing hepatic injury and accelerating hepatocyte proliferation after carbon tetrachloride(CCl4) treatment.METHODS: C57 BL/6 mic...AIM: To assess the effects of dihydromyricetin(DHM) as a hepatoprotective candidate in reducing hepatic injury and accelerating hepatocyte proliferation after carbon tetrachloride(CCl4) treatment.METHODS: C57 BL/6 mice were used in this study. Mice were orally administered with DHM(150 mg/kg) for 4 d after CCl4 treatment. Serum and liver tissue samples were collected on days 1, 2, 3, 5 and 7 after CCl4 treatment. The anti-inflammatory effect of DHM was assessed directly by hepatic histology detection and indirectly by serum levels of aspartate aminotransferase(AST), alanine aminotransferase(ALT), albumin, and superoxide dismutase(SOD). Inflammatory cytokines, such as interleukin(IL)-1β, IL-6 and tumor necrosis factor-α(TNF-α), were detected using ELISA kits. Proliferating cell nuclear antigen(PCNA) staining was used to evaluate the role of DHM in promoting hepatocyte proliferation. Hepatocyte apoptosis wasmeasured by TUNEL assay.Furthermore,apoptosis proteins Caspases-3,6,8,and 9 were detected by Western blot.SP600125 were used to confirm whether DHM regulated liver regeneration through JNK/TNF-αpathways.RESULTS:DHM showed a strong anti-inflammatory effect on CCl4-induced liver injury in mice.DHM could significantly decrease serum ALT,AST,IL-1β,IL-6 and TNF-αand increase serum albumin,SOD and liver SOD compared to the control group after CCl4 treatment(P<0.05).PCNA results indicated that DHM could significantly increase the number of PCNA positive cells compared to the control(348.9±56.0 vs 107.1±31.4,P<0.01).TUNEL assay showed that DHM dramatically reduced the number of apoptotic cells after CCl4 treatment compared to the control(365.4±99.4 vs 90.5±13.8,P<0.01).Caspase activity detection showed that DHM could reduce the activities of Caspases-8,3,6 and 9 compared to the control(P<0.05).The results of Western blot showed that DHM increased the expression of JNK and decreased TNF-αexpression.However,DHM could not affect TNF-αexpression after SP600125 treatment.Furthermore,DHM could significantly improve the survival rate of acute liver failure(ALF)mice(73.3%vs 20.0%,P<0.0001),and SP600125 could inhibit the effect of DHM.CONCLUSION:These findings demonstrate that DHM alleviates CCl4-induced liver injury,suggesting that DHM is a promising candidate for reversing liver injury and ALF.展开更多
AIM To investigate whether Dihydromyricetin(DHM) inhibits cell proliferation and promotes apoptosis by downregulating Notch1 expression.METHODS The correlation between Notch1 and Hes1(a Notch1 target molecule) express...AIM To investigate whether Dihydromyricetin(DHM) inhibits cell proliferation and promotes apoptosis by downregulating Notch1 expression.METHODS The correlation between Notch1 and Hes1(a Notch1 target molecule) expression in hepatoma samples was confirmed by q RT-PCR. In addition, MTT assays, flow cytometry and TUNEL analysis showed that DHM possessed strong anti-tumor properties, evidenced not only by reduced cell proliferation but also by enhanced apoptosis in QGY7701 and Hep G2 hepatocellular carcinoma(HCC) cells. The expressions of Notch1, Hes1, Bcl-2 and Bax were determined by Western blot.RESULTS Among the tested samples(n = 64), the expression levels of Notch1(75% of patients) and Hes1(79.7% of patients) m RNA in tumor tissues were higher than in the normal liver tissues. There was a negative correlation between the expression of Notch1 and the degree of differentiation and positively correlated with the Alpha Fetal Protein concentration. The viability of HCC cells treated with DHM was significantly inhibited in a dose and time-dependent manner. Apoptosis was induced in Hep G2 and QGY7701 cell lines following 24 h of DHM treatment. After treatment with DHM, the protein expression of Notch1 was downregulated, the apoptosis-related protein Bax was upregulated and Bcl2 was downregulated. Notch1 si RNA further enhanced the anti-tumor properties of DHM. CONCLUSION Notch1 is involved in the development of HCC and DHM inhibits cell proliferation and promotes apoptosis by down-regulating the expression of Notch1.展开更多
Vine tea has been used as an herbal tea by several ethnic minorities for hundreds of years in China.Flavonoids,a kind of indispensable component in a variety of nutraceutical,pharmaceutical and cosmetic applications,a...Vine tea has been used as an herbal tea by several ethnic minorities for hundreds of years in China.Flavonoids,a kind of indispensable component in a variety of nutraceutical,pharmaceutical and cosmetic applications,are identified to be the major metabolites and bioactive ingredients in vine tea.Interestingly,vine tea exhibits a wide range of significant bioactivities including anti-oxidant,anti-inflammatory,anti-tumor,antidiabetic,neuroprotective and other activities,but no toxicity.These bioactivities,to some extent,enrich the understanding about the role of vine tea in disease prevention and therapy.The health benefits of vine tea,particularly dihydromyricetin and myricetin,are widely investigated.However,there is currently no comprehensive review available on vine tea.Therefore,this report summarizes the most recent studies investigating bioactive constituents,pharmacological effects and possible mechanisms of vine tea,which will provide a better understanding about the health benefits and preclinical assessment of novel application of vine tea.展开更多
BACKGROUND Chronic liver injury(CLI) is now a worldwide disease. However, there is no effective treatment. Pyroptosis plays an essential role in CLI. Dihydromyricetin(DHM) resists oxidation and protects the liver. We ...BACKGROUND Chronic liver injury(CLI) is now a worldwide disease. However, there is no effective treatment. Pyroptosis plays an essential role in CLI. Dihydromyricetin(DHM) resists oxidation and protects the liver. We hypothesize that the beneficial effect of DHM on CLI is related to its effect on the expression of pyroptosisrelated molecules. Therefore, we studied the influence of DHM on CLI and pyroptosis.AIM To study the role of pyroptosis in the pathogenesis of CLI and the therapeutic mechanism of DHM.METHODS Thirty-two mice were randomly divided into four groups: The control group was injected with olive oil, the carbon tetrachloride(CCl4) group was injected with CCl4, the vehicle group was injected with hydroxypropyl-β-cyclodextrin while injecting CCl4 and the DHM group was injected with DHM while injecting CCl4. After four weeks of treatment, liver tissues from the mice were stained with hematoxylin and eosin, and oil red O. Blood was collected from the angular vein for serological analysis. The severity of CLI was estimated. Some liver tissue was sampled for immunohistochemistry, Western blotting and quantitative reverse transcription PCR to observe the changes in pyroptosis-related molecules.RESULTS Serum total cholesterol, low density lipoprotein, aspartate aminotransferase(AST) and alanine aminotransferase(ALT) in the CCl4 group were higher than those in the control group, and serum total cholesterol, low density lipoprotein, AST and ALT in the DHM group were lower than those in the vehicle group. Hematoxylin and eosin and oil red O staining showed that there were more lipid droplets in the CCl4 group than in the control group, and there were fewer lipid droplets in the DHM group than in the vehicle group. Western blotting showed that the expression of the pyroptosis-related molecules caspase-1, NOD-, LRR-and pyrin domain-containing 3(NLRP3) and gasdermin D(GSDMD)-N in the CCl4 group was higher than that in the control group, while expression of these proteins in the DHM group was lower than that in the vehicle group. Quantitative reverse transcription PCR results showed that the expression of the pyroptosis-related genes caspase-1, NLRP3, GSDMD and interleukin-1β(IL-1β) in the CCl4 group was higher than that in the control group, while there was no significant change in NLRP3 and caspase-1 expression in the DHM group compared with that in the vehicle group, and the expression of GSDMD and IL-1β was decreased.CONCLUSION DHM improves CCl4-induced CLI and regulates the pyroptosis pathway in hepatocytes. DHM may be a potential therapeutic agent for CLI.展开更多
Background:Vine tea from fermented Ampelopsis grossedentata leaves has been used as a herbal tea and folk medicine in the southern region of China for hundreds of years.The aim of this investigation was to analyze the...Background:Vine tea from fermented Ampelopsis grossedentata leaves has been used as a herbal tea and folk medicine in the southern region of China for hundreds of years.The aim of this investigation was to analyze the total flavonoids found in vine tea,including three bioactive flavonoids,and the total phenolic contents in the aqueous methanol extracts of 10 vine tea samples.In addition,this study also aimed to examine the antioxidant activity of dihydromyricetin and vine tea’s flavonoid-rich extract.Methods:The total flavonoids and total phenolic content assay of extracts from vine tea were performed by ultraviolet-visible spectroscopy and epoch microplate spectrophotometer,respectively.Three bioactive flavonoids were quantified simultaneously using high performance liquid chromatography.The antioxidant activity of dihydromyricetin and vine tea’s flavonoid-rich extract was evaluated in vitro using six different methods.Results:Vine tea contained a large number of flavonoids,with dihydromyricetin as its main constituent.The flavonoid-rich extract exhibited a significant scavenging effect on superoxide anion radicals,and on 3-ethylbenzthiazoline-6-sulphonic acid and 1,1-diphenyl-2-picrylhydrazyl radicals.It also possessed definite activity in lipid peroxidation inhibition,ferric reduction,and the moderation of Fe2+ion chelation ability.There was a significant negative correlation between dihydromyricetin content and antioxidant activity in the vine tea samples,including superoxide anion radical scavenging activity(P=−0.754,P<0.05),lipid peroxidation inhibition activity(P=−0.759,P<0.05),ferric-reducing antioxidant power(P=−0.843,P<0.01),respectively.Dihydromyricetin played a dominant role in the antioxidant activities of the flavonoid-rich extract.Conclusion:Vine tea’s flavonoid-rich extract could be used as a new antioxidant source to safeguard against oxidative stress.展开更多
Ampelopsis grossedentata(Hand-Mazz) W. T. wang is a typical edible plant with important physiological health functions. Dihydromyricetin is the main active ingredient of A. grossedentata, which has good pharmacologica...Ampelopsis grossedentata(Hand-Mazz) W. T. wang is a typical edible plant with important physiological health functions. Dihydromyricetin is the main active ingredient of A. grossedentata, which has good pharmacological effects such as antibacterial, anti-inflammatory, anti-oxidation, anti-tumor and liver protecting effects. This paper summarized the research progress on the extraction materials, extraction methods and separation and purification processes of dihydromyricetin in A. grossedentata to understand the best experimental method of dihydromyricetin, aiming to provide an important theoretical basis for the comprehensive development and utilization of dihydromyricetin.展开更多
Cassia ferruginea (Schrad.) DC.(Fabaceae) is a native plant widely used as an ornamental tree and for the restoration of degraded areas in Brazil. Although no previous ethnopharmacological or chemical information was ...Cassia ferruginea (Schrad.) DC.(Fabaceae) is a native plant widely used as an ornamental tree and for the restoration of degraded areas in Brazil. Although no previous ethnopharmacological or chemical information was available, bioprospecting studies on this species found out a high concentration of dihydromyricetin (DHM) in different parts of the plant. DHM, also known as (+)-ampelopsin, is a flavonoid that presents important pharmacological and biological activities, including anticancer, cardioprotective and hepatoprotective effects. The aim of this study was to develop and validate a simple method for the determination of DHM in hydroethanolic extracts of C. ferruginea by quantitative 1H NMR (qHNMR) using the internal standard approach. The parameters as linearity, specificity and selectivity, accuracy, precision (repeatability and intermediate) and robustness showed satisfactory results. Five ethanolic extracts from different stages and organs of C. ferruginea were investigated, the DHM concentration ranges from 52.08 ± 3.95 to 201.83 ± 4.71 mg/g. The proposed method appears to be a suitable tool for fast quality control of herbal extracts, with no need of tedious sample preparation and chromatographic separations increasing the efficiency of natural products analysis. This is the first study reporting the presence and determination of DHM in Cassia ferruginea (Schrad.) DC.展开更多
The geometry of myricetin and dihydromyricetin was optimized by DMol3 program based on DFT. The optimized structures have been proved to be stable by real frequencies obtained.The parameters of geometry optimization,v...The geometry of myricetin and dihydromyricetin was optimized by DMol3 program based on DFT. The optimized structures have been proved to be stable by real frequencies obtained.The parameters of geometry optimization,vibration frequencies,atomic charges,thermodynamics,Fukui functions,and frontier molecular orbital had been obtained. The thermodynamic properties of 2 molecules were fitted and the relationship between the thermodynamic function and temperature was obtained. The correlation coefficient of the obtained equations is larger than 0.99,indicating that the function is approximately linear in the whole temperature range. The calculated results showed the O–H molecular structures were the main group affecting the antioxidant activity and the oxygen in the benzene ring is the electrophilic reaction site. Ortho-OH on the benzene formed intramolecular hydrogen bonds,which contributed to the stability of the benzene ring. It may be the active site for the occurrence of the reaction.展开更多
To provide a reference basis for reasonable development and utilization of Ampelopsis grossedentata resource and its application in production,we added 0.025%,0.05% and 0.1% dihydromyricetin(DMY)extracted from A.gross...To provide a reference basis for reasonable development and utilization of Ampelopsis grossedentata resource and its application in production,we added 0.025%,0.05% and 0.1% dihydromyricetin(DMY)extracted from A.grossedentata into the basal diets of Yaoshan chickens,and studied the effects of DMY on visceral organ indexes and digestive enzyme activities of 40-day-old healthy Yaoshan chickens.Supplementation of DMY in basal diets influenced visceral organ weight,organ index,length and digestive enzyme activity of small intestine of chickens.Among them,0.05% DMY was the most appropriate volume for supplementation.Compared with control group(without DMY),0.05% DMY reduced lung weight and index(P>0.05),significantly reduced liver weight(P<0.05),extremely reduced stomach weight and stomach index(P<0.01),increased activity of amylase in small intestine(P>0.05),significantly increased length of small intestine and activity of protease(P<0.05).Supplementation of 0.05% DMY reduced the visceral organ weight and organ index,enhanced digestion and absorption ability of gastrointestinal tract,thereby improving feed utilization rate and promoting the growth of chickens.展开更多
Objective: To investigate the effect and potential mechanism of dihydromyricetin(Dmy) on H9C2 cell proliferation, apoptosis, and autophagy. Methods: H9C2 cells were randomly divided into 7 groups, namely control, mode...Objective: To investigate the effect and potential mechanism of dihydromyricetin(Dmy) on H9C2 cell proliferation, apoptosis, and autophagy. Methods: H9C2 cells were randomly divided into 7 groups, namely control, model, EV(empty p CDH-CMV-MCS-EF1-Cop GFP-T2A-Puro vector), IV(circHIPK3 interference), Dmy(50 μmol/L), Dmy+IV, and Dmy+EV groups. Cell proliferation and apoptosis were detected by cell counting kit-8 assay and flow cytometry, respectivley. Western blot was used to evaluate the levels of light chain 3 Ⅱ/Ⅰ(LC3Ⅱ/Ⅰ), phospho-phosphoinositide 3-kinase(p-PI3K), protein kinase B(p-AKT), and phospho-mammalian target of rapamycin(p-mTOR). The level of circHIPK3 was determined using reverse transcriptase polymerase chain reaction. Electron microscopy was used to observe autophagosomes in H9C2 cells. Results: Compared to H9C2 cells, the expression of circHIPK in H9C2 hypoxia model cells increased significantly(P<0.05). Compared to the control group, the cell apoptosis and autophagosomes increased, cell proliferation rate decreased significantly, and the expression of LC3Ⅱ/Ⅰ significantly increased(all P<0.05). Compared to the model group, the rate of apoptosis and autophagosomes in IV, Dmy, and Dmy+IV group decreased, the cell proliferation rate increased, and the expression of LC3Ⅱ/Ⅰ decreased significantly(all P<0.05). Compared to the control group, the expressions of p-PI3K, p-AKT, and p-mTOR in the model group significantly reduced(P<0.05), whereas after treatment with Dmy and sh-circHIPK3, the above situation was reversed(P<0.05). Conclusion: Dmy plays a protective role in H9C2 cells by inhibiting circHIPK expression and cell apoptosis and autophagy, and the mechanism may be related to PI3K/AKT/mTOR pathway.展开更多
OBJECTIVE: To investigate the efficacy of dihydromyricetin(DMY) on nasopharyngeal carcinoma(NPC) cell proliferation, apoptosis and to reveal the underlying mechanism in vitro experiments.METHODS: The CNE-2 cell line w...OBJECTIVE: To investigate the efficacy of dihydromyricetin(DMY) on nasopharyngeal carcinoma(NPC) cell proliferation, apoptosis and to reveal the underlying mechanism in vitro experiments.METHODS: The CNE-2 cell line was treated with different concentrations of DMY and the effects of DMY on cell viability and proliferation were evaluated using cell counting kit-8(CCK-8) assay and plate colony formation assay. Cellular apoptosis was detected by flow cytometry following Annexin V fluorescein isothiocyanate/propidine iodide staining.Nuclei morphology was observed under a fluorescence microscope following Hoechst 333258 staining. The expression of phosphorylated inhibitor of nuclear factor kappa-B kinase subunit beta(p-IKKβ), phosphorylated inhibitor of nuclear factor kappa-B kinase subunit alpha(p-IKKα), inhibitor of nuclear factor kappa-B alpha(IκB-α), nuclear factor kappa-B(NF-κB)/p65 was examined by Western blot analysis and the nuclear translocation of NF-κB/p65 was observed using a confocal laser scanning microscopy.RESULTS: DMY inhibited the proliferative capability and colony formation of NPC CNE-2 cells. Meanwhile, DMY induced apoptosis of CNE-2 cells in a dose and time-dependent manner via upregulating B-cell lymphoma-2 associated X, but downregulating B-cell lymphoma-2 and pro-caspase-3. Importantly, we found that DMY suppressed tumor necrosis factor alpha(TNF-α)-mediated NF-κB activation via inhibiting p-IKKβ, p-IKKα and blocking NF-κB subunit p65.CONCLUSION: Our experiments demonstrated that DMY had significant antiproliferative and apoptosisinducing effects on CNE-2 cells. Additionally, DMY promoted inactivation of p-IKKβ, p-IKKα,and blocked the nuclear translocation of NF-κB subunit p65. These results suggest that DMY may be an important therapeutic approach for NPC.展开更多
The harsh microenvironment in wound(HMW)remains a major obstacle to chronic wound healing.Although a series of bioactive materials have been developed,few of them are multi-functional and able to accelerate wound heal...The harsh microenvironment in wound(HMW)remains a major obstacle to chronic wound healing.Although a series of bioactive materials have been developed,few of them are multi-functional and able to accelerate wound healing via precisely remodeling the HMW.Herein,a series of dihydromyricetin(DHM)-incorporated multilayer nanofibers(termed DQHP-n,n=0,2,6 and 10)are fabricated using a layer-by-layer(LBL)self-assembly technique.The average diameters of DQHP-n significantly increase from 0.30±0.16μm to 0.84±0.28μm(P<0.05)along with the n value increased from 0 to 10,the tensile strength of that is also significantly improved from 1.12±0.15 MPa to 2.16±0.30 MPa(P<0.05),and the water contact angle of that significantly decreases from 129.1±1.5°to 76.6±3.9°(P<0.05).The DQHP-n are found to be biocompatible,in which DQHP-6 promoted cell migration through activation of the epithelial–mesenchymal transformation(EMT)pathway and reconstruction of the HMW by stopping bleeding,killing bacteria,eliminating inflammation,and scavenging reactive oxygen species(ROS).The in vivo evaluation is carried out via an E.coli-infected rat skin regeneration model.The DQHP-6 group demonstrates the best effect,as it healed up to 98.5±1.0%of the wound area at day 15.DQHP-6 differentially regulates the mRNA expressions of several cytokines(FGF2,PDGF,IL-1α,IL-6,IL10,and TGF-β),which ends to reductions of total inflammatory cells(CD45^(+) cells)and M1 macrophages(CD80^(+) and CD86^(+) cells),proliferation of host cell(Ki67^(+) cells),and enhancement of collagen synthesis.In conclusion,DQHP-6 exhibits multifunctional properties for HMW,and can serve as a promising wound dressing for clinical transformation.展开更多
基金Supported by The National Natural Science Foundation of China,No.81041099Natural Science Foundation of Guangdong Province,China,No.S2011010003750
文摘AIM:To investigate the effects of dihydromyricetin(DHM)on the migration and invasion of human hepatic cancer cells.METHODS:The hepatoma cell lines SK-Hep-1 and MHCC97L were used in this study.The cells were cultured in RPIM-1640 medium supplemented with 10%fetal bovine serum at 37℃in a humidified 5%CO2incubator.DHM was dissolved in dimethyl sulfoxide and diluted to various concentrations in medium before applying to cells.MTT assays were performed to measure the viability of the cells after DHM treatment.Wound healing and Boyden transwell assays were used to assess cancer cell motility.The invasive capacity of cancer cells was measured using Matrigel-coated transwell chambers.Matrix metalloproteinase(MMP)-2/9 activity was examined by fluorescence analysis.Western blot was carried out to analyze the expression of MMP-2,MMP-9,p-38,JNK,ERK1/2 and PKC-δproteins.All data were analyzed by Student’s t tests in GraphPad prism 5.0software and are presented as mean±SD.RESULTS:DHM was found to strongly inhibit the migration of the hepatoma cell lines SK-Hep-1(without DHM,24 h:120±8μmol/L vs 100μmol/L DHM,24h:65±10μmol/L,P<0.001)and MHCC97L(without DHM,24 h:126±7μmol/L vs 100μmol/L DHM,24h:74±6μmol/L,P<0.001).The invasive capacity of the cells was reduced by DHM treatment(SK-Hep-1cells without DHM,24 h:67±4μmol/L vs 100μmol/L DHM,24 h:9±3μmol/L,P<0.001;MHCC97L cells without DHM,24 h:117±8μmol/L vs 100μmol/L DHM,24 h:45±2μmol/L,P<0.001).MMP2/9 activity was also inhibited by DHM exposure(SK-Hep-1 cells without DHM,24 h:600±26μmol/L vs 100μmol/L DHM,24 h:100±6μmol/L,P<0.001;MHCC97L cells without DHM,24 h:504±32μmol/L vs 100μmol/L DHM 24 h:156±10μmol/L,P<0.001).Western blot analysis showed that DHM decreased the expression level of MMP-9 but had little effect on MMP-2.Further investigation indicated that DHM markedly reduced the phosphorylation levels of p38,ERK1/2 and JNK in a concentration-dependent manner but had no impact on the total protein levels.In addition,PKC-δprotein,a key protein in the regulation of MMP family protein expression,was up-regulated with DHM treatment.CONCLUSION:These findings demonstrate that DHM inhibits the migration and invasion of hepatoma cells and may serve as a potential candidate agent for the prevention of HCC metastasis.
基金Supported by Initial Fund of Guangdong Medical College,No.XB1338the Medical Research Fund of Guangdong Province,No.B2014306the Research Fund of Guangdong Medical College,No.M2013024
文摘AIM: To assess the effects of dihydromyricetin(DHM) as a hepatoprotective candidate in reducing hepatic injury and accelerating hepatocyte proliferation after carbon tetrachloride(CCl4) treatment.METHODS: C57 BL/6 mice were used in this study. Mice were orally administered with DHM(150 mg/kg) for 4 d after CCl4 treatment. Serum and liver tissue samples were collected on days 1, 2, 3, 5 and 7 after CCl4 treatment. The anti-inflammatory effect of DHM was assessed directly by hepatic histology detection and indirectly by serum levels of aspartate aminotransferase(AST), alanine aminotransferase(ALT), albumin, and superoxide dismutase(SOD). Inflammatory cytokines, such as interleukin(IL)-1β, IL-6 and tumor necrosis factor-α(TNF-α), were detected using ELISA kits. Proliferating cell nuclear antigen(PCNA) staining was used to evaluate the role of DHM in promoting hepatocyte proliferation. Hepatocyte apoptosis wasmeasured by TUNEL assay.Furthermore,apoptosis proteins Caspases-3,6,8,and 9 were detected by Western blot.SP600125 were used to confirm whether DHM regulated liver regeneration through JNK/TNF-αpathways.RESULTS:DHM showed a strong anti-inflammatory effect on CCl4-induced liver injury in mice.DHM could significantly decrease serum ALT,AST,IL-1β,IL-6 and TNF-αand increase serum albumin,SOD and liver SOD compared to the control group after CCl4 treatment(P<0.05).PCNA results indicated that DHM could significantly increase the number of PCNA positive cells compared to the control(348.9±56.0 vs 107.1±31.4,P<0.01).TUNEL assay showed that DHM dramatically reduced the number of apoptotic cells after CCl4 treatment compared to the control(365.4±99.4 vs 90.5±13.8,P<0.01).Caspase activity detection showed that DHM could reduce the activities of Caspases-8,3,6 and 9 compared to the control(P<0.05).The results of Western blot showed that DHM increased the expression of JNK and decreased TNF-αexpression.However,DHM could not affect TNF-αexpression after SP600125 treatment.Furthermore,DHM could significantly improve the survival rate of acute liver failure(ALF)mice(73.3%vs 20.0%,P<0.0001),and SP600125 could inhibit the effect of DHM.CONCLUSION:These findings demonstrate that DHM alleviates CCl4-induced liver injury,suggesting that DHM is a promising candidate for reversing liver injury and ALF.
基金Supported by The National Natural Science Foundation of China,No.81041099Natural Science Foundation of Guangdong Province,China,No.S2011010003750
文摘AIM To investigate whether Dihydromyricetin(DHM) inhibits cell proliferation and promotes apoptosis by downregulating Notch1 expression.METHODS The correlation between Notch1 and Hes1(a Notch1 target molecule) expression in hepatoma samples was confirmed by q RT-PCR. In addition, MTT assays, flow cytometry and TUNEL analysis showed that DHM possessed strong anti-tumor properties, evidenced not only by reduced cell proliferation but also by enhanced apoptosis in QGY7701 and Hep G2 hepatocellular carcinoma(HCC) cells. The expressions of Notch1, Hes1, Bcl-2 and Bax were determined by Western blot.RESULTS Among the tested samples(n = 64), the expression levels of Notch1(75% of patients) and Hes1(79.7% of patients) m RNA in tumor tissues were higher than in the normal liver tissues. There was a negative correlation between the expression of Notch1 and the degree of differentiation and positively correlated with the Alpha Fetal Protein concentration. The viability of HCC cells treated with DHM was significantly inhibited in a dose and time-dependent manner. Apoptosis was induced in Hep G2 and QGY7701 cell lines following 24 h of DHM treatment. After treatment with DHM, the protein expression of Notch1 was downregulated, the apoptosis-related protein Bax was upregulated and Bcl2 was downregulated. Notch1 si RNA further enhanced the anti-tumor properties of DHM. CONCLUSION Notch1 is involved in the development of HCC and DHM inhibits cell proliferation and promotes apoptosis by down-regulating the expression of Notch1.
文摘Vine tea has been used as an herbal tea by several ethnic minorities for hundreds of years in China.Flavonoids,a kind of indispensable component in a variety of nutraceutical,pharmaceutical and cosmetic applications,are identified to be the major metabolites and bioactive ingredients in vine tea.Interestingly,vine tea exhibits a wide range of significant bioactivities including anti-oxidant,anti-inflammatory,anti-tumor,antidiabetic,neuroprotective and other activities,but no toxicity.These bioactivities,to some extent,enrich the understanding about the role of vine tea in disease prevention and therapy.The health benefits of vine tea,particularly dihydromyricetin and myricetin,are widely investigated.However,there is currently no comprehensive review available on vine tea.Therefore,this report summarizes the most recent studies investigating bioactive constituents,pharmacological effects and possible mechanisms of vine tea,which will provide a better understanding about the health benefits and preclinical assessment of novel application of vine tea.
基金Supported by National Natural Science Foundation of China,No. 81873769Beijing Municipal Natural Science Foundation,No. 7162098。
文摘BACKGROUND Chronic liver injury(CLI) is now a worldwide disease. However, there is no effective treatment. Pyroptosis plays an essential role in CLI. Dihydromyricetin(DHM) resists oxidation and protects the liver. We hypothesize that the beneficial effect of DHM on CLI is related to its effect on the expression of pyroptosisrelated molecules. Therefore, we studied the influence of DHM on CLI and pyroptosis.AIM To study the role of pyroptosis in the pathogenesis of CLI and the therapeutic mechanism of DHM.METHODS Thirty-two mice were randomly divided into four groups: The control group was injected with olive oil, the carbon tetrachloride(CCl4) group was injected with CCl4, the vehicle group was injected with hydroxypropyl-β-cyclodextrin while injecting CCl4 and the DHM group was injected with DHM while injecting CCl4. After four weeks of treatment, liver tissues from the mice were stained with hematoxylin and eosin, and oil red O. Blood was collected from the angular vein for serological analysis. The severity of CLI was estimated. Some liver tissue was sampled for immunohistochemistry, Western blotting and quantitative reverse transcription PCR to observe the changes in pyroptosis-related molecules.RESULTS Serum total cholesterol, low density lipoprotein, aspartate aminotransferase(AST) and alanine aminotransferase(ALT) in the CCl4 group were higher than those in the control group, and serum total cholesterol, low density lipoprotein, AST and ALT in the DHM group were lower than those in the vehicle group. Hematoxylin and eosin and oil red O staining showed that there were more lipid droplets in the CCl4 group than in the control group, and there were fewer lipid droplets in the DHM group than in the vehicle group. Western blotting showed that the expression of the pyroptosis-related molecules caspase-1, NOD-, LRR-and pyrin domain-containing 3(NLRP3) and gasdermin D(GSDMD)-N in the CCl4 group was higher than that in the control group, while expression of these proteins in the DHM group was lower than that in the vehicle group. Quantitative reverse transcription PCR results showed that the expression of the pyroptosis-related genes caspase-1, NLRP3, GSDMD and interleukin-1β(IL-1β) in the CCl4 group was higher than that in the control group, while there was no significant change in NLRP3 and caspase-1 expression in the DHM group compared with that in the vehicle group, and the expression of GSDMD and IL-1β was decreased.CONCLUSION DHM improves CCl4-induced CLI and regulates the pyroptosis pathway in hepatocytes. DHM may be a potential therapeutic agent for CLI.
基金This work was supported by Traditional Chinese Medicine Science and Technology Development Project of Shandong Province(No.2019-0649)Fundamental Research Funds for the Central Universities(No.2018-BUCMXJKY001).
文摘Background:Vine tea from fermented Ampelopsis grossedentata leaves has been used as a herbal tea and folk medicine in the southern region of China for hundreds of years.The aim of this investigation was to analyze the total flavonoids found in vine tea,including three bioactive flavonoids,and the total phenolic contents in the aqueous methanol extracts of 10 vine tea samples.In addition,this study also aimed to examine the antioxidant activity of dihydromyricetin and vine tea’s flavonoid-rich extract.Methods:The total flavonoids and total phenolic content assay of extracts from vine tea were performed by ultraviolet-visible spectroscopy and epoch microplate spectrophotometer,respectively.Three bioactive flavonoids were quantified simultaneously using high performance liquid chromatography.The antioxidant activity of dihydromyricetin and vine tea’s flavonoid-rich extract was evaluated in vitro using six different methods.Results:Vine tea contained a large number of flavonoids,with dihydromyricetin as its main constituent.The flavonoid-rich extract exhibited a significant scavenging effect on superoxide anion radicals,and on 3-ethylbenzthiazoline-6-sulphonic acid and 1,1-diphenyl-2-picrylhydrazyl radicals.It also possessed definite activity in lipid peroxidation inhibition,ferric reduction,and the moderation of Fe2+ion chelation ability.There was a significant negative correlation between dihydromyricetin content and antioxidant activity in the vine tea samples,including superoxide anion radical scavenging activity(P=−0.754,P<0.05),lipid peroxidation inhibition activity(P=−0.759,P<0.05),ferric-reducing antioxidant power(P=−0.843,P<0.01),respectively.Dihydromyricetin played a dominant role in the antioxidant activities of the flavonoid-rich extract.Conclusion:Vine tea’s flavonoid-rich extract could be used as a new antioxidant source to safeguard against oxidative stress.
基金Supported by Project of Guilin Science and Technology Bureau(20140105-12)
文摘Ampelopsis grossedentata(Hand-Mazz) W. T. wang is a typical edible plant with important physiological health functions. Dihydromyricetin is the main active ingredient of A. grossedentata, which has good pharmacological effects such as antibacterial, anti-inflammatory, anti-oxidation, anti-tumor and liver protecting effects. This paper summarized the research progress on the extraction materials, extraction methods and separation and purification processes of dihydromyricetin in A. grossedentata to understand the best experimental method of dihydromyricetin, aiming to provide an important theoretical basis for the comprehensive development and utilization of dihydromyricetin.
文摘Cassia ferruginea (Schrad.) DC.(Fabaceae) is a native plant widely used as an ornamental tree and for the restoration of degraded areas in Brazil. Although no previous ethnopharmacological or chemical information was available, bioprospecting studies on this species found out a high concentration of dihydromyricetin (DHM) in different parts of the plant. DHM, also known as (+)-ampelopsin, is a flavonoid that presents important pharmacological and biological activities, including anticancer, cardioprotective and hepatoprotective effects. The aim of this study was to develop and validate a simple method for the determination of DHM in hydroethanolic extracts of C. ferruginea by quantitative 1H NMR (qHNMR) using the internal standard approach. The parameters as linearity, specificity and selectivity, accuracy, precision (repeatability and intermediate) and robustness showed satisfactory results. Five ethanolic extracts from different stages and organs of C. ferruginea were investigated, the DHM concentration ranges from 52.08 ± 3.95 to 201.83 ± 4.71 mg/g. The proposed method appears to be a suitable tool for fast quality control of herbal extracts, with no need of tedious sample preparation and chromatographic separations increasing the efficiency of natural products analysis. This is the first study reporting the presence and determination of DHM in Cassia ferruginea (Schrad.) DC.
基金supported by the Science and Technology Project of Guangdong Province(2016B010122032)Natural Science Foundation of Guangdong Province(05300953)
文摘The geometry of myricetin and dihydromyricetin was optimized by DMol3 program based on DFT. The optimized structures have been proved to be stable by real frequencies obtained.The parameters of geometry optimization,vibration frequencies,atomic charges,thermodynamics,Fukui functions,and frontier molecular orbital had been obtained. The thermodynamic properties of 2 molecules were fitted and the relationship between the thermodynamic function and temperature was obtained. The correlation coefficient of the obtained equations is larger than 0.99,indicating that the function is approximately linear in the whole temperature range. The calculated results showed the O–H molecular structures were the main group affecting the antioxidant activity and the oxygen in the benzene ring is the electrophilic reaction site. Ortho-OH on the benzene formed intramolecular hydrogen bonds,which contributed to the stability of the benzene ring. It may be the active site for the occurrence of the reaction.
文摘To provide a reference basis for reasonable development and utilization of Ampelopsis grossedentata resource and its application in production,we added 0.025%,0.05% and 0.1% dihydromyricetin(DMY)extracted from A.grossedentata into the basal diets of Yaoshan chickens,and studied the effects of DMY on visceral organ indexes and digestive enzyme activities of 40-day-old healthy Yaoshan chickens.Supplementation of DMY in basal diets influenced visceral organ weight,organ index,length and digestive enzyme activity of small intestine of chickens.Among them,0.05% DMY was the most appropriate volume for supplementation.Compared with control group(without DMY),0.05% DMY reduced lung weight and index(P>0.05),significantly reduced liver weight(P<0.05),extremely reduced stomach weight and stomach index(P<0.01),increased activity of amylase in small intestine(P>0.05),significantly increased length of small intestine and activity of protease(P<0.05).Supplementation of 0.05% DMY reduced the visceral organ weight and organ index,enhanced digestion and absorption ability of gastrointestinal tract,thereby improving feed utilization rate and promoting the growth of chickens.
基金Supported by Science Foundation of Education Department of Shaanxi Provincial Government (No.19JK0890)Natural Science Foundation of Xizang (Tibet) Autonomous Region (No.XZ202001ZR0089G)+1 种基金Major Training Project of Xizang Minzu University (Nos.18MDZ03 and 20MDT03)Funded Project of Young Scholar Cultivation Program of Xizang Minzu University (No.21MDX04)。
文摘Objective: To investigate the effect and potential mechanism of dihydromyricetin(Dmy) on H9C2 cell proliferation, apoptosis, and autophagy. Methods: H9C2 cells were randomly divided into 7 groups, namely control, model, EV(empty p CDH-CMV-MCS-EF1-Cop GFP-T2A-Puro vector), IV(circHIPK3 interference), Dmy(50 μmol/L), Dmy+IV, and Dmy+EV groups. Cell proliferation and apoptosis were detected by cell counting kit-8 assay and flow cytometry, respectivley. Western blot was used to evaluate the levels of light chain 3 Ⅱ/Ⅰ(LC3Ⅱ/Ⅰ), phospho-phosphoinositide 3-kinase(p-PI3K), protein kinase B(p-AKT), and phospho-mammalian target of rapamycin(p-mTOR). The level of circHIPK3 was determined using reverse transcriptase polymerase chain reaction. Electron microscopy was used to observe autophagosomes in H9C2 cells. Results: Compared to H9C2 cells, the expression of circHIPK in H9C2 hypoxia model cells increased significantly(P<0.05). Compared to the control group, the cell apoptosis and autophagosomes increased, cell proliferation rate decreased significantly, and the expression of LC3Ⅱ/Ⅰ significantly increased(all P<0.05). Compared to the model group, the rate of apoptosis and autophagosomes in IV, Dmy, and Dmy+IV group decreased, the cell proliferation rate increased, and the expression of LC3Ⅱ/Ⅰ decreased significantly(all P<0.05). Compared to the control group, the expressions of p-PI3K, p-AKT, and p-mTOR in the model group significantly reduced(P<0.05), whereas after treatment with Dmy and sh-circHIPK3, the above situation was reversed(P<0.05). Conclusion: Dmy plays a protective role in H9C2 cells by inhibiting circHIPK expression and cell apoptosis and autophagy, and the mechanism may be related to PI3K/AKT/mTOR pathway.
基金Supported by the National Natural Science Foundation of China (Regulation and Mechanism of Bone Marrow Mesenchymal Stem Cells on the Characteristics of Nasopharyngeal Carcinoma Stem Cells, No.81272434)the Medical Research Fund of Guangdong Province (Effect of Berberine on Growth of Transplanted Tumor of Nasopharyngeal Carcinoma in Mice Based on JAK/STAT3 Signaling Pathway, No.A2016431)。
文摘OBJECTIVE: To investigate the efficacy of dihydromyricetin(DMY) on nasopharyngeal carcinoma(NPC) cell proliferation, apoptosis and to reveal the underlying mechanism in vitro experiments.METHODS: The CNE-2 cell line was treated with different concentrations of DMY and the effects of DMY on cell viability and proliferation were evaluated using cell counting kit-8(CCK-8) assay and plate colony formation assay. Cellular apoptosis was detected by flow cytometry following Annexin V fluorescein isothiocyanate/propidine iodide staining.Nuclei morphology was observed under a fluorescence microscope following Hoechst 333258 staining. The expression of phosphorylated inhibitor of nuclear factor kappa-B kinase subunit beta(p-IKKβ), phosphorylated inhibitor of nuclear factor kappa-B kinase subunit alpha(p-IKKα), inhibitor of nuclear factor kappa-B alpha(IκB-α), nuclear factor kappa-B(NF-κB)/p65 was examined by Western blot analysis and the nuclear translocation of NF-κB/p65 was observed using a confocal laser scanning microscopy.RESULTS: DMY inhibited the proliferative capability and colony formation of NPC CNE-2 cells. Meanwhile, DMY induced apoptosis of CNE-2 cells in a dose and time-dependent manner via upregulating B-cell lymphoma-2 associated X, but downregulating B-cell lymphoma-2 and pro-caspase-3. Importantly, we found that DMY suppressed tumor necrosis factor alpha(TNF-α)-mediated NF-κB activation via inhibiting p-IKKβ, p-IKKα and blocking NF-κB subunit p65.CONCLUSION: Our experiments demonstrated that DMY had significant antiproliferative and apoptosisinducing effects on CNE-2 cells. Additionally, DMY promoted inactivation of p-IKKβ, p-IKKα,and blocked the nuclear translocation of NF-κB subunit p65. These results suggest that DMY may be an important therapeutic approach for NPC.
基金This work was financially supported by the Fellowship of China National Postdoctoral Program for Innovative Talants(BX20220240)the Improvement Project for Theranostic Ability on Difficulty Miscellaneous Disease(Tumor)from National Health Commission of China(ZLYNXM202006)+1 种基金the Chinese Central Special Fund for Local Science and Technology Development of Hubei Province(2018ZYYD023)the Science and Technology Department of Hubei Province Key Project(2018ACA159).
文摘The harsh microenvironment in wound(HMW)remains a major obstacle to chronic wound healing.Although a series of bioactive materials have been developed,few of them are multi-functional and able to accelerate wound healing via precisely remodeling the HMW.Herein,a series of dihydromyricetin(DHM)-incorporated multilayer nanofibers(termed DQHP-n,n=0,2,6 and 10)are fabricated using a layer-by-layer(LBL)self-assembly technique.The average diameters of DQHP-n significantly increase from 0.30±0.16μm to 0.84±0.28μm(P<0.05)along with the n value increased from 0 to 10,the tensile strength of that is also significantly improved from 1.12±0.15 MPa to 2.16±0.30 MPa(P<0.05),and the water contact angle of that significantly decreases from 129.1±1.5°to 76.6±3.9°(P<0.05).The DQHP-n are found to be biocompatible,in which DQHP-6 promoted cell migration through activation of the epithelial–mesenchymal transformation(EMT)pathway and reconstruction of the HMW by stopping bleeding,killing bacteria,eliminating inflammation,and scavenging reactive oxygen species(ROS).The in vivo evaluation is carried out via an E.coli-infected rat skin regeneration model.The DQHP-6 group demonstrates the best effect,as it healed up to 98.5±1.0%of the wound area at day 15.DQHP-6 differentially regulates the mRNA expressions of several cytokines(FGF2,PDGF,IL-1α,IL-6,IL10,and TGF-β),which ends to reductions of total inflammatory cells(CD45^(+) cells)and M1 macrophages(CD80^(+) and CD86^(+) cells),proliferation of host cell(Ki67^(+) cells),and enhancement of collagen synthesis.In conclusion,DQHP-6 exhibits multifunctional properties for HMW,and can serve as a promising wound dressing for clinical transformation.
