Objective To investigate the anti-oxidant and anti-inflammatory effects of ethanol extract of Polygala sibirica L.var megalopha Fr.(EEP)on RAW264.7 mouse macrophages.Methods RAW264.7 cells were pretreated with 0–200&...Objective To investigate the anti-oxidant and anti-inflammatory effects of ethanol extract of Polygala sibirica L.var megalopha Fr.(EEP)on RAW264.7 mouse macrophages.Methods RAW264.7 cells were pretreated with 0–200µg/mL EEP or vehicle for 2 h prior to exposure to 1µg/mL lipopolysaccharide(LPS)for 24 h.Nitric oxide(NO)and prostaglandin(PGE2)production were determined by Griess reagent and enzyme-linked immunosorbent assay(ELISA),respectively.The mRNA levels of inducible nitric oxide synthase(iNOS),cyclooxygenase-2(COX-2),tumor necrosis factorα(TNF-α),interleukin-1beta(IL-1β),and IL-6 were determined using reverse transcription polymerase chain reaction(RT-PCR).Western blot assay was used to determine the protein expressions of iNOS,COX-2,phosphorylation of extracellular regulated protein kinases(ERK1/2),c-Jun N-terminal kinase(JNK),inhibitory subunit of nuclear factor Kappa B alpha(IκB-α)and p38.Immunofluorescence was used to observe the nuclear expression of nuclear factor-κB p65(NF-κB p65).Additionally,the anti-oxidant potential of EEP was evaluated by reactive oxygen species(ROS)production and the activities of catalase(CAT)and superoxide dismutase(SOD).The 2,2-diphenyl-1-picrylhydrazyl(DPPH),hydroxyl(OH),superoxide anion(O2−)radical and nitrite scavenging activity were also measured.Results The total polyphenol and flavonoid contents of EEP were 23.50±2.16 mg gallic acid equivalent/100 g and 43.78±3.81 mg rutin equivalent/100 g.With EEP treatment(100 and 150µg/mL),there was a notable decrease in NO and PGE2 production induced by LPS in RAW264.7 cells by downregulation of iNOS and COX-2 mRNA and protein expressions(P<0.01 or P<0.05).Furthermore,with EEP treatment(150µg/mL),there was a decrease in the mRNA expression levels of TNF-α,IL-1βand IL-6,as well as in the phosphorylation of ERK,JNK and p38 mitogen-activated protein kinase(MAPK,P<0.01 or P<0.05),by blocking the nuclear translocation of NF-κB p65 in LPS-stimulated cells.In addition,EEP(100 and 150µg/mL)led to an increase in the anti-oxidant enzymes activity of SOD and CAT,with a concomitant decrease in ROS production(P<0.01 or P<0.05).EEP also indicated the DPPH,OH,O2−radical and nitrite scavenging activity.Conclusion EEP inhibited inflammatory responses in activated macrophages through blocking MAPK/NF-κB pathway and protected against oxidative stress.展开更多
AIM:To investigate the chemical constituents of the roots of Polygala sibirica L.(Polygalaceae) METHOD:The isolation was performed by solvent extraction and various chromatographic techniques,including silica gel,Seph...AIM:To investigate the chemical constituents of the roots of Polygala sibirica L.(Polygalaceae) METHOD:The isolation was performed by solvent extraction and various chromatographic techniques,including silica gel,Sephadex LH-20,ODS,semi-preparative HPLC,and preparative TLC.The chemical structures were elucidated based on extensive spectroscopic analysis,including HR-ESI-MS and 1D- and 2D-NMR spectroscopic data.RESULTS:A total of sixteen compounds,including five xanthones(5,7–10),five saccharide esters(1,3,4,12,13),two flavonoids(14,16),two triterpenoids(11,15),one phenylpropanoid(6),and one benzophenone glycoside(2) were isolated.Their structures were determined as sibiricose A7(1),sibiriphenone A(2),polygalatenoside A(3),polygalatenoside C(4),lancerin(5),3,4,5-trimethoxycinnamic acid(6),6-hydroxy-1,2,3,7-tetramethoxyxanthone(7),1,3,7-trihydroxy-2-methoxyxanthone(8),onjixanthone II(9),1,2,3,6,7-pentamethoxyxanthone(10),presenegenin(11),3'-O-3,4,5-trimethoxycinnamoyl-6-O-4-methoxy benzoyl sucrose(12),tenuifoliside C(13),5,3'-dihydroxy-7,4'-dimethoxyflavonol-3-O-β-D-glucopyranoside(14),tenuifolin(15),and rhamnetin 3-O-β-D-glucopyranoside(16).CONCLUSION:Compounds 1 and 2 are two new compounds from P.sibirica.展开更多
文摘Objective To investigate the anti-oxidant and anti-inflammatory effects of ethanol extract of Polygala sibirica L.var megalopha Fr.(EEP)on RAW264.7 mouse macrophages.Methods RAW264.7 cells were pretreated with 0–200µg/mL EEP or vehicle for 2 h prior to exposure to 1µg/mL lipopolysaccharide(LPS)for 24 h.Nitric oxide(NO)and prostaglandin(PGE2)production were determined by Griess reagent and enzyme-linked immunosorbent assay(ELISA),respectively.The mRNA levels of inducible nitric oxide synthase(iNOS),cyclooxygenase-2(COX-2),tumor necrosis factorα(TNF-α),interleukin-1beta(IL-1β),and IL-6 were determined using reverse transcription polymerase chain reaction(RT-PCR).Western blot assay was used to determine the protein expressions of iNOS,COX-2,phosphorylation of extracellular regulated protein kinases(ERK1/2),c-Jun N-terminal kinase(JNK),inhibitory subunit of nuclear factor Kappa B alpha(IκB-α)and p38.Immunofluorescence was used to observe the nuclear expression of nuclear factor-κB p65(NF-κB p65).Additionally,the anti-oxidant potential of EEP was evaluated by reactive oxygen species(ROS)production and the activities of catalase(CAT)and superoxide dismutase(SOD).The 2,2-diphenyl-1-picrylhydrazyl(DPPH),hydroxyl(OH),superoxide anion(O2−)radical and nitrite scavenging activity were also measured.Results The total polyphenol and flavonoid contents of EEP were 23.50±2.16 mg gallic acid equivalent/100 g and 43.78±3.81 mg rutin equivalent/100 g.With EEP treatment(100 and 150µg/mL),there was a notable decrease in NO and PGE2 production induced by LPS in RAW264.7 cells by downregulation of iNOS and COX-2 mRNA and protein expressions(P<0.01 or P<0.05).Furthermore,with EEP treatment(150µg/mL),there was a decrease in the mRNA expression levels of TNF-α,IL-1βand IL-6,as well as in the phosphorylation of ERK,JNK and p38 mitogen-activated protein kinase(MAPK,P<0.01 or P<0.05),by blocking the nuclear translocation of NF-κB p65 in LPS-stimulated cells.In addition,EEP(100 and 150µg/mL)led to an increase in the anti-oxidant enzymes activity of SOD and CAT,with a concomitant decrease in ROS production(P<0.01 or P<0.05).EEP also indicated the DPPH,OH,O2−radical and nitrite scavenging activity.Conclusion EEP inhibited inflammatory responses in activated macrophages through blocking MAPK/NF-κB pathway and protected against oxidative stress.
基金Program for Changjiang Scholar and Innovative Team in University(Grant No.985-2-063-112)Program for New Century Excellent Talents in University(Grant No.985-2-102-113).
基金financially supported by the National Key Technology R&D Program"New Drug Innovation"of China(Nos.2012ZX09301002-002-002,2012ZX09304-005)special funds for scientific research on traditional Chinese medicine(No.201307002)National Science Fund for Excellent Young Scholars(No.81222051)
文摘AIM:To investigate the chemical constituents of the roots of Polygala sibirica L.(Polygalaceae) METHOD:The isolation was performed by solvent extraction and various chromatographic techniques,including silica gel,Sephadex LH-20,ODS,semi-preparative HPLC,and preparative TLC.The chemical structures were elucidated based on extensive spectroscopic analysis,including HR-ESI-MS and 1D- and 2D-NMR spectroscopic data.RESULTS:A total of sixteen compounds,including five xanthones(5,7–10),five saccharide esters(1,3,4,12,13),two flavonoids(14,16),two triterpenoids(11,15),one phenylpropanoid(6),and one benzophenone glycoside(2) were isolated.Their structures were determined as sibiricose A7(1),sibiriphenone A(2),polygalatenoside A(3),polygalatenoside C(4),lancerin(5),3,4,5-trimethoxycinnamic acid(6),6-hydroxy-1,2,3,7-tetramethoxyxanthone(7),1,3,7-trihydroxy-2-methoxyxanthone(8),onjixanthone II(9),1,2,3,6,7-pentamethoxyxanthone(10),presenegenin(11),3'-O-3,4,5-trimethoxycinnamoyl-6-O-4-methoxy benzoyl sucrose(12),tenuifoliside C(13),5,3'-dihydroxy-7,4'-dimethoxyflavonol-3-O-β-D-glucopyranoside(14),tenuifolin(15),and rhamnetin 3-O-β-D-glucopyranoside(16).CONCLUSION:Compounds 1 and 2 are two new compounds from P.sibirica.
