Objective:To evaluate the effects of Capsosiphon fulvescens(C.fulvescens)ethanolic extract on inflammation in lipopolysaccharide(LPS)-induced RAW296.7 macrophages.Methods:The protective effects of C.fulvescens ethanol...Objective:To evaluate the effects of Capsosiphon fulvescens(C.fulvescens)ethanolic extract on inflammation in lipopolysaccharide(LPS)-induced RAW296.7 macrophages.Methods:The protective effects of C.fulvescens ethanolic extract on LPS-induced inflammation in RAW264.7 macrophages were assessed using biochemical analysis,including enzyme-linked immunosorbent assay,quantitative reverse transcription-polymerase chain reaction,and Western blot analysis.To examine reactive oxygen species(ROS)production,flow cytometry analysis,and immunofluorescence staining were used.Furthermore,the modulatory effect of C.fulvescens ethanolic extract on NF-κB activation was investigated.Results:C.fulvescens ethanolic extract significantly attenuated LPS-induced levels of pro-inflammatory cytokines and notably reduced the secretion and mRNA levels of LPS-mediated matrix metalloproteinases.In addition,C.fulvescens ethanolic extract decreased ROS production and suppressed the TLR4/NF-κB signaling pathway.Conclusions:C.fulvescens ethanolic extract alleviates inflammation as well as oxidative stress by modulating the TLR4/NF-κB signaling in LPS-induced RAW264.7 macrophages.C.fulvescens can be used as a potential therapeutic agent to suppress inflammation and oxidative stress-associated diseases.展开更多
Objective:To investigate the effects of an ethanol extract of Kalopanax septemlobus(Thunb.)Koidz.leaf(EEKS) on cell proliferation in human hepatocellular carcinoma cells and its mechanisms of action.Methods:Cells were...Objective:To investigate the effects of an ethanol extract of Kalopanax septemlobus(Thunb.)Koidz.leaf(EEKS) on cell proliferation in human hepatocellular carcinoma cells and its mechanisms of action.Methods:Cells were treated with EEKS and subsequently analyzed for cell proliferation and flow cytometry analysis.Expressions of cell cycle regulators were determined by reverse transcriptase polymerase chain reaction analysis and Western blotting,and activation of eyclin-associaled kinases studied using kinase assays.Results:The EEKS suppressed cell proliferation in both HepG2 and Hep3 B cells,but showed a more sensitive anli-proliferative activity in HepG2 cells.Flow cytometry analysis revealed an association between the growth inhibitory effect of EEKS and with G_1 phase cell cycle arrest in HepG2 cells,along with the dephosphorylation of retinoblastoma protein(pRB) and enhanced binding of pRB with the E2 F transcription factor family proteins.Treatment with EEKS also increased the expression of cyclin-dependent kinase(CDK) inhibitors,such as p21WAF1/CIP1 and p27KIP1.without any noticeable changes in G_1 cyclins and CDKs(except for a slight decrease in CDK4).Treatment of HepG2 cells with EEKS also increased the binding of p21 and p27 with CDK4 and CDK6.which was paralleled by a marked decrease in the cyclin D- and cyclin E-associated kinase activities.Conclusions:Overall,our findings suggest that EEKS may be an effective treatment for liver cancer through suppression of cancer cell proliferation via G_1,cell cycle arrest Further studies arc required to identify the active compounds in EEKS.展开更多
Objective:To investigate whether the ethanol extract of Chondracanthus tenellus(Harvey)Hommersand,a type of red algae,could exhibit anti-inflammatory potential in lipopolysaccharide(LPS)-stimulated macrophages.Methods...Objective:To investigate whether the ethanol extract of Chondracanthus tenellus(Harvey)Hommersand,a type of red algae,could exhibit anti-inflammatory potential in lipopolysaccharide(LPS)-stimulated macrophages.Methods:The ethanol extract of Chondracanthus tenellus was applied to 100 ng/mL LPS-stimulated RAW 264.7 cells,and cell viability,phagocytic ability,levels of pro-inflammatory factors,and the production of reactive oxygen species were measured.To identify the underlying mechanism of the ethanol extract of Chondracanthus tenellus,the expression of inflammation-regulated genes was estimated.Results:The ethanol extract of Chondracanthus tenellus had no cytotoxic effect at concentrations below 300μg/mL,and reduced the LPS-induced production of inflammatory mediators including nitric oxide(NO)and prostaglandin E2.Furthermore,the extract markedly suppressed the expression of inducible NO synthase and cyclooxygenase-2,as well as the production of reactive oxygen species.The LPS-induced up-regulation of pro-inflammatory cytokines was attenuated by treatment with the ethanol extract of Chondracanthus tenellus,reducing their extracellular secretion.The Chondracanthus tenellus extract also inhibited LPS-mediated activation of nuclear factor-kappa B(NF-κB).In addition,the phosphorylation of mitogen activated protein kinases(MAPKs)and phosphatidylinositol 3 kinase(PI3K)/Akt was markedly increased by LPS,which was significantly abolished by the Chondracanthus tenellus extract.Conclusions:Our findings indicate that the ethanol extract of Chondracanthus tenellus exhibited potential anti-inflammatory and antioxidant effects through downregulating the NF-κB,MAPKs,and PI3K/Akt signaling pathways in LPS stimulated RAW 264.7 macrophages.tenellus extract.Conclusions:Our findings indicate that the ethanol extract of Chondracanthus tenellus exhibited potential anti-inflammatory and antioxidant effects through downregulating the NF-κB,MAPKs,and PI3K/Akt signaling pathways in LPS stimulated RAW 264.7 macrophages.展开更多
Objective:To investigate the effect of honokiol on oxidative damage in HaCaT human keratinocytes.Methods:HaCaT cells were exposed to hydrogen peroxide(H_(2)O_(2)),following pretreatment with various concentrations of ...Objective:To investigate the effect of honokiol on oxidative damage in HaCaT human keratinocytes.Methods:HaCaT cells were exposed to hydrogen peroxide(H_(2)O_(2)),following pretreatment with various concentrations of honokiol.The alleviating effects of honokiol on HaCaT cell viability and cell death,reactive oxygen species(ROS)production,DNA damage,mitochondrial dynamics,and inhibition of adenosine triphoaphate production against H_(2)O_(2)were investigated.Western blotting analysis was used to analyze the expression levels of specific proteins.Results:Honokiol suppressed H_(2)O_(2)-induced cytotoxicity and DNA damage by blocking abnormal ROS accumulation.Honokiol also prevented apoptosis by inhibiting loss of mitochondrial membrane potential and release of cytochrome c from the mitochondria into the cytosol,decreasing the Bax/Bcl-2 ratio,and reducing the activity of caspase-3 in H_(2)O_(2)-stimulated HaCaT cells.In addition,honokiol attenuated H_(2)O_(2)-induced reduction of adenosine triphosphate content,and activation of AMP-activated protein kinase(AMPK)was markedly promoted by honokiol in H_(2)O_(2)-stimulated cells.Importantly,the anti-apoptosis and anti-proliferative activity of honokiol against H_(2)O_(2)was further enhanced by adding an activator of AMPK,indicating that honokiol activated AMPK in HaCaT keratinocytes to protect against oxidative damage.Conclusions:The present results indicate that honokiol may be useful as a potential therapeutic agent against various oxidative stress-related skin diseases.展开更多
Objective:To investigate whether ethanol extracts of Hizikia fusiforme could induce apoptosis in human prostate cancer PC3 cells.Methods:Cell viability was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetr...Objective:To investigate whether ethanol extracts of Hizikia fusiforme could induce apoptosis in human prostate cancer PC3 cells.Methods:Cell viability was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide.Apoptosis and mitochondrial membrane potential(MMP)were measured using flow cytometry in PC3 cells.DNA damage was assessed by nuclear staining and DNA fragmentation assay.Expressions of apoptosis-associated proteins were determined by Western blotting assays.Activities of caspase-3,-8,and-9 were determined by colorimetric assay.Moreover,intracellular reactive oxygen species(ROS)generation was detected using a flow cytometer and fluorescence microscope.Results:Treatment of PC3 cells with ethanol extracts of Hizikia fusiforme inhibited proliferation,which was associated with induction of apoptosis,and accompanied by increased expression of Fas,Fas-ligand(Fas L),Bax and t Bid,and decreased expression of Bcl-2.In addition,ethanol extracts of Hizikia fusiforme reduced c-Flip expression and activated caspase-8,-9 and-3,resulting in an increase in poly(ADP-ribose)polymerase(PARP)cleavage.However,in the presence of a pan-caspase inhibitor,ethanol extracts of Hizikia fusiforme-mediated growth inhibition and apoptosis were significantly attenuated.Ethanol extracts of Hizikia fusiforme also destroyed the integrity of mitochondria due to the loss of MMP,leading to cytosolic release of cytochrome c.Moreover,the levels of ROS were markedly increased by treatment with ethanol extracts of Hizikia fusiforme,which was significantly suppressed by the ROS scavenger N-acetyl-L-cysteine.Further investigation of whether ethanol extracts of Hizikia fusiforme-induced apoptosis was related to the generation of ROS was conducted and the results showed that N-acetyl-L-cysteine fully blocked ethanol extracts of Hizikia fusiforme-induced apoptotic events including loss of MMP,activation of caspase-3,the cytosolic release of cytochrome c and cytotoxicity.Conclusions:Ethanol extracts of Hizikia fusiforme have chemopreventive potential via induction of ROS-dependent apoptosis.Therefore,ethanol extracts of Hizikia fusiforme may be useful for developing effective and selective natural sources to inhibit cancer cell proliferation.展开更多
Experimental stroke research commonly employs focal cerebral ischemic rat models （Bederson et al., 1986a; Longa et al., 1989）. In human patients, ischemic stroke typically results from thrombotic or embolic occlusio...Experimental stroke research commonly employs focal cerebral ischemic rat models （Bederson et al., 1986a; Longa et al., 1989）. In human patients, ischemic stroke typically results from thrombotic or embolic occlusion of a major cerebral artery, usually the mid- dle cerebral artery （MCA）. Experimental focal cerebral ischemia models have been employed to mimic human stroke （Durukan and Tatlisumak, 2007）. Rodent models of focal cerebral ischemia that do not require craniotomy have been developed using intraluminal suture occlusion of the MCA （MCA occlusion, MCAO） （Rosamond et al., 2008）. Furthermore, mouse MCAO models have been wide- ly used and extended to genetic studies of cell death or recovery mechanisms （Liu and McCullough, 2011）. Genetically engineered mouse stroke models are particularly useful for evaluation of isch- emic pathophysiology and the design of new prophylactic, neuro- protective, and therapeutic agents and interventions （Armstead et al., 2010）. During the past two decades, MCAO surgical techniques have been developed that do not reveal surgical techniques for mouse MCAO model engineering. Therefore, we compared MCAO surgical methods in rats and mice.展开更多
Objective:To investigate whether ethanol extracts of Chondracanthus tenellus(EECT)could improve immunomodulatory property of murine monocyte/macrophage RAW 264.7 cells.Methods:Cell viability,phagocytic ability,and nit...Objective:To investigate whether ethanol extracts of Chondracanthus tenellus(EECT)could improve immunomodulatory property of murine monocyte/macrophage RAW 264.7 cells.Methods:Cell viability,phagocytic ability,and nitric oxide were measured.The levels of prostaglandin E2 and cytokines were determined using enzyme-linked immunosorbent assay kits.Expression of immunoregulatory response protein was detected by Western blotting assay.Results:As the concentration of EECT increased,the morphology of the cells changed to a typical active macrophage shape,and the phagocytic activity increased significantly.EECT also effectively enhanced the production and secretion of immunomodulatory mediators,such as nitric oxide and prostaglandin E2,and cytokines.In addition,compared with the control group,EECT markedly stimulated the expression of Toll-like receptor 4(TLR4)and myeloid differentiation factor 88,one of the TLR4 adapter molecules.Furthermore,EECT promoted the nucleus translocation of nuclear factor-kappa B(NF-κB)by increasing the phosphorylation and degradation of the inhibitor of NF-κB-α,indicating activation of the NF-κB signaling pathway.Meanwhile,similar trends were found in cells treated with lipopolysaccharide as a positive control.Conclusions:Taken together,the results indicate that EECT has an immunomodulatory effect by increasing the production of immunomodulatory mediators and cytokines through activation of the TLR4/NF-κB signaling pathway.EECT could be used as a potential candidate for medication or dietary supplements to increase immune activity.展开更多
基金funded by Korea Institute of Marine Science&Technology Promotion(KIMST)funded by the Ministry of Oceans and Fisheries,Korea(20220488).
文摘Objective:To evaluate the effects of Capsosiphon fulvescens(C.fulvescens)ethanolic extract on inflammation in lipopolysaccharide(LPS)-induced RAW296.7 macrophages.Methods:The protective effects of C.fulvescens ethanolic extract on LPS-induced inflammation in RAW264.7 macrophages were assessed using biochemical analysis,including enzyme-linked immunosorbent assay,quantitative reverse transcription-polymerase chain reaction,and Western blot analysis.To examine reactive oxygen species(ROS)production,flow cytometry analysis,and immunofluorescence staining were used.Furthermore,the modulatory effect of C.fulvescens ethanolic extract on NF-κB activation was investigated.Results:C.fulvescens ethanolic extract significantly attenuated LPS-induced levels of pro-inflammatory cytokines and notably reduced the secretion and mRNA levels of LPS-mediated matrix metalloproteinases.In addition,C.fulvescens ethanolic extract decreased ROS production and suppressed the TLR4/NF-κB signaling pathway.Conclusions:C.fulvescens ethanolic extract alleviates inflammation as well as oxidative stress by modulating the TLR4/NF-κB signaling in LPS-induced RAW264.7 macrophages.C.fulvescens can be used as a potential therapeutic agent to suppress inflammation and oxidative stress-associated diseases.
基金supported by Basic Science Research Program through the National Research Foundation of Korea grant funded by the Korea government(2015RLA2A2A01004633 and 2014RIAIA1008460)
文摘Objective:To investigate the effects of an ethanol extract of Kalopanax septemlobus(Thunb.)Koidz.leaf(EEKS) on cell proliferation in human hepatocellular carcinoma cells and its mechanisms of action.Methods:Cells were treated with EEKS and subsequently analyzed for cell proliferation and flow cytometry analysis.Expressions of cell cycle regulators were determined by reverse transcriptase polymerase chain reaction analysis and Western blotting,and activation of eyclin-associaled kinases studied using kinase assays.Results:The EEKS suppressed cell proliferation in both HepG2 and Hep3 B cells,but showed a more sensitive anli-proliferative activity in HepG2 cells.Flow cytometry analysis revealed an association between the growth inhibitory effect of EEKS and with G_1 phase cell cycle arrest in HepG2 cells,along with the dephosphorylation of retinoblastoma protein(pRB) and enhanced binding of pRB with the E2 F transcription factor family proteins.Treatment with EEKS also increased the expression of cyclin-dependent kinase(CDK) inhibitors,such as p21WAF1/CIP1 and p27KIP1.without any noticeable changes in G_1 cyclins and CDKs(except for a slight decrease in CDK4).Treatment of HepG2 cells with EEKS also increased the binding of p21 and p27 with CDK4 and CDK6.which was paralleled by a marked decrease in the cyclin D- and cyclin E-associated kinase activities.Conclusions:Overall,our findings suggest that EEKS may be an effective treatment for liver cancer through suppression of cancer cell proliferation via G_1,cell cycle arrest Further studies arc required to identify the active compounds in EEKS.
基金This research was a part of the project titled‘Omics based on fishery disease control technology development and industrialization(20150242)’,funded by the Ministry of Oceans and Fisheries,Republic of Korea。
文摘Objective:To investigate whether the ethanol extract of Chondracanthus tenellus(Harvey)Hommersand,a type of red algae,could exhibit anti-inflammatory potential in lipopolysaccharide(LPS)-stimulated macrophages.Methods:The ethanol extract of Chondracanthus tenellus was applied to 100 ng/mL LPS-stimulated RAW 264.7 cells,and cell viability,phagocytic ability,levels of pro-inflammatory factors,and the production of reactive oxygen species were measured.To identify the underlying mechanism of the ethanol extract of Chondracanthus tenellus,the expression of inflammation-regulated genes was estimated.Results:The ethanol extract of Chondracanthus tenellus had no cytotoxic effect at concentrations below 300μg/mL,and reduced the LPS-induced production of inflammatory mediators including nitric oxide(NO)and prostaglandin E2.Furthermore,the extract markedly suppressed the expression of inducible NO synthase and cyclooxygenase-2,as well as the production of reactive oxygen species.The LPS-induced up-regulation of pro-inflammatory cytokines was attenuated by treatment with the ethanol extract of Chondracanthus tenellus,reducing their extracellular secretion.The Chondracanthus tenellus extract also inhibited LPS-mediated activation of nuclear factor-kappa B(NF-κB).In addition,the phosphorylation of mitogen activated protein kinases(MAPKs)and phosphatidylinositol 3 kinase(PI3K)/Akt was markedly increased by LPS,which was significantly abolished by the Chondracanthus tenellus extract.Conclusions:Our findings indicate that the ethanol extract of Chondracanthus tenellus exhibited potential anti-inflammatory and antioxidant effects through downregulating the NF-κB,MAPKs,and PI3K/Akt signaling pathways in LPS stimulated RAW 264.7 macrophages.tenellus extract.Conclusions:Our findings indicate that the ethanol extract of Chondracanthus tenellus exhibited potential anti-inflammatory and antioxidant effects through downregulating the NF-κB,MAPKs,and PI3K/Akt signaling pathways in LPS stimulated RAW 264.7 macrophages.
文摘Objective:To investigate the effect of honokiol on oxidative damage in HaCaT human keratinocytes.Methods:HaCaT cells were exposed to hydrogen peroxide(H_(2)O_(2)),following pretreatment with various concentrations of honokiol.The alleviating effects of honokiol on HaCaT cell viability and cell death,reactive oxygen species(ROS)production,DNA damage,mitochondrial dynamics,and inhibition of adenosine triphoaphate production against H_(2)O_(2)were investigated.Western blotting analysis was used to analyze the expression levels of specific proteins.Results:Honokiol suppressed H_(2)O_(2)-induced cytotoxicity and DNA damage by blocking abnormal ROS accumulation.Honokiol also prevented apoptosis by inhibiting loss of mitochondrial membrane potential and release of cytochrome c from the mitochondria into the cytosol,decreasing the Bax/Bcl-2 ratio,and reducing the activity of caspase-3 in H_(2)O_(2)-stimulated HaCaT cells.In addition,honokiol attenuated H_(2)O_(2)-induced reduction of adenosine triphosphate content,and activation of AMP-activated protein kinase(AMPK)was markedly promoted by honokiol in H_(2)O_(2)-stimulated cells.Importantly,the anti-apoptosis and anti-proliferative activity of honokiol against H_(2)O_(2)was further enhanced by adding an activator of AMPK,indicating that honokiol activated AMPK in HaCaT keratinocytes to protect against oxidative damage.Conclusions:The present results indicate that honokiol may be useful as a potential therapeutic agent against various oxidative stress-related skin diseases.
基金a part of the project titled‘Omics based on fishery disease control technology development and industrialization(20150242)’‘Development of functional food products with natural materials derived from marine resources(2017-0377)’funded by the Ministry of Oceans and Fisheries,Republic of Korea.
文摘Objective:To investigate whether ethanol extracts of Hizikia fusiforme could induce apoptosis in human prostate cancer PC3 cells.Methods:Cell viability was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide.Apoptosis and mitochondrial membrane potential(MMP)were measured using flow cytometry in PC3 cells.DNA damage was assessed by nuclear staining and DNA fragmentation assay.Expressions of apoptosis-associated proteins were determined by Western blotting assays.Activities of caspase-3,-8,and-9 were determined by colorimetric assay.Moreover,intracellular reactive oxygen species(ROS)generation was detected using a flow cytometer and fluorescence microscope.Results:Treatment of PC3 cells with ethanol extracts of Hizikia fusiforme inhibited proliferation,which was associated with induction of apoptosis,and accompanied by increased expression of Fas,Fas-ligand(Fas L),Bax and t Bid,and decreased expression of Bcl-2.In addition,ethanol extracts of Hizikia fusiforme reduced c-Flip expression and activated caspase-8,-9 and-3,resulting in an increase in poly(ADP-ribose)polymerase(PARP)cleavage.However,in the presence of a pan-caspase inhibitor,ethanol extracts of Hizikia fusiforme-mediated growth inhibition and apoptosis were significantly attenuated.Ethanol extracts of Hizikia fusiforme also destroyed the integrity of mitochondria due to the loss of MMP,leading to cytosolic release of cytochrome c.Moreover,the levels of ROS were markedly increased by treatment with ethanol extracts of Hizikia fusiforme,which was significantly suppressed by the ROS scavenger N-acetyl-L-cysteine.Further investigation of whether ethanol extracts of Hizikia fusiforme-induced apoptosis was related to the generation of ROS was conducted and the results showed that N-acetyl-L-cysteine fully blocked ethanol extracts of Hizikia fusiforme-induced apoptotic events including loss of MMP,activation of caspase-3,the cytosolic release of cytochrome c and cytotoxicity.Conclusions:Ethanol extracts of Hizikia fusiforme have chemopreventive potential via induction of ROS-dependent apoptosis.Therefore,ethanol extracts of Hizikia fusiforme may be useful for developing effective and selective natural sources to inhibit cancer cell proliferation.
基金supported by the 2013 Inje University Research Grant
文摘Experimental stroke research commonly employs focal cerebral ischemic rat models （Bederson et al., 1986a; Longa et al., 1989）. In human patients, ischemic stroke typically results from thrombotic or embolic occlusion of a major cerebral artery, usually the mid- dle cerebral artery （MCA）. Experimental focal cerebral ischemia models have been employed to mimic human stroke （Durukan and Tatlisumak, 2007）. Rodent models of focal cerebral ischemia that do not require craniotomy have been developed using intraluminal suture occlusion of the MCA （MCA occlusion, MCAO） （Rosamond et al., 2008）. Furthermore, mouse MCAO models have been wide- ly used and extended to genetic studies of cell death or recovery mechanisms （Liu and McCullough, 2011）. Genetically engineered mouse stroke models are particularly useful for evaluation of isch- emic pathophysiology and the design of new prophylactic, neuro- protective, and therapeutic agents and interventions （Armstead et al., 2010）. During the past two decades, MCAO surgical techniques have been developed that do not reveal surgical techniques for mouse MCAO model engineering. Therefore, we compared MCAO surgical methods in rats and mice.
基金This research was a part of the project titled‘Omics based on fishery disease control technology development and industrialization(20150242)’funded by the Ministry of Oceans and Fisheries,Republic of Korea.
文摘Objective:To investigate whether ethanol extracts of Chondracanthus tenellus(EECT)could improve immunomodulatory property of murine monocyte/macrophage RAW 264.7 cells.Methods:Cell viability,phagocytic ability,and nitric oxide were measured.The levels of prostaglandin E2 and cytokines were determined using enzyme-linked immunosorbent assay kits.Expression of immunoregulatory response protein was detected by Western blotting assay.Results:As the concentration of EECT increased,the morphology of the cells changed to a typical active macrophage shape,and the phagocytic activity increased significantly.EECT also effectively enhanced the production and secretion of immunomodulatory mediators,such as nitric oxide and prostaglandin E2,and cytokines.In addition,compared with the control group,EECT markedly stimulated the expression of Toll-like receptor 4(TLR4)and myeloid differentiation factor 88,one of the TLR4 adapter molecules.Furthermore,EECT promoted the nucleus translocation of nuclear factor-kappa B(NF-κB)by increasing the phosphorylation and degradation of the inhibitor of NF-κB-α,indicating activation of the NF-κB signaling pathway.Meanwhile,similar trends were found in cells treated with lipopolysaccharide as a positive control.Conclusions:Taken together,the results indicate that EECT has an immunomodulatory effect by increasing the production of immunomodulatory mediators and cytokines through activation of the TLR4/NF-κB signaling pathway.EECT could be used as a potential candidate for medication or dietary supplements to increase immune activity.
