Objective:To demonstrate the effect of dieckol from Eisenia bicyclis on osteoclastogenesis using RAW 264.7 cells.Methods:Murine macrophage RAW 264.7 cells were subjected to dieckol treatment,followed by treatment with...Objective:To demonstrate the effect of dieckol from Eisenia bicyclis on osteoclastogenesis using RAW 264.7 cells.Methods:Murine macrophage RAW 264.7 cells were subjected to dieckol treatment,followed by treatment with receptor activator of nuclear factor kappa-B ligand(RANKL)to induce osteoclastogenesis.Tartrate-resistant acid phosphatase(TRAP)activity was examined using a TRAP activity kit.Western blotting analysis was conducted to examine the level of osteoclast-related factors,including TRAP and calcitonin receptor(CTR),transcriptional factors,including c-Fos,c-Jun,and nuclear factor of activated T cells cytoplasmic 1(NFATc1),nuclear factor kappa-B(NF-κB),extracellular signal-regulated kinase(ERK),and c-Jun N-terminal kinase(JNK).Immunofluorescence staining was conducted to examine the expression of c-Fos,c-Jun,and NFATc1.Results:Among the four phlorotannin compounds present in Eisenia bicyclis,dieckol significantly hindered osteoclast differentiation and expression of RANKL-induced TRAP and CTR.In addition,dieckol downregulated the expression levels of c-Fos,c-Jun,NFATc1,ERK,and JNK,and suppressed NF-κB signaling.Conclusions:Dieckol can suppress RANKL-induced osteoclastogenesis.Therefore,it has therapeutic potential in treating osteoclastogenesis-associated diseases.展开更多
Curcumfa phaeocaulis Valeton is used in traditional Chinese medicines for the treatment of blood-related disorders such as blood stasis and inflammation.Our previous studies have found that sesquiterpenes were mainly ...Curcumfa phaeocaulis Valeton is used in traditional Chinese medicines for the treatment of blood-related disorders such as blood stasis and inflammation.Our previous studies have found that sesquiterpenes were mainly isolated from C.wenyujin and C.phaeocaulis,while diarylheptanoids were the major compounds from C.kwangsiensis.And most of the isolated compounds exhibited remarkable展开更多
Objective:To demonstrate the anti-inflammatory activity of Brassica napus L.hydrosols(BNH)in lipopolysaccharide(LPS)-stimulated RAW 264.7 cells.Methods:Composition analysis of BNH was conducted via gas chromatography-...Objective:To demonstrate the anti-inflammatory activity of Brassica napus L.hydrosols(BNH)in lipopolysaccharide(LPS)-stimulated RAW 264.7 cells.Methods:Composition analysis of BNH was conducted via gas chromatography-mass spectrometry after BNH were extracted.The nitric oxide(NO)production was measured using the Griess assay.Prostaglandin E2(PGE2)production was evaluated with enzyme-linked immunosorbent assay.The effects of BNH on LPS-induced pro-inflammatory enzymes including inducible nitric oxide synthase(iNOS)and cyclooxygenase-2(COX-2)were evaluated using Western blot analysis.Furthermore,phosphorylation of nuclear factor-kappa B(N F-k B)and nuclear translocation of N F-k B p65 were evaluated with Western blot analysis and immunofluorescence staining,respectively.Results:Compared with LPS-stimulated cells,BNH markedly decreased the generation of NO and PGE_(2)in LPS-stimulated RAW 264.7 cells(P<0.01 or P<0.05).Moreover,BNH inhibited protein levels of iNOS and COX-2(P<0.01).Phosphorylation of NF-k B and nuclear translocation of NF-k B p65 was significantly inhibited by BNH(P<0.01 or P<0.05).Conclusion:The anti-inflammatory activities of BNH were mediated via blockage of the N F-k B signaling pathways in LPS-stimulated RAW 264.7 cells.展开更多
Two immunomodulatory polysaccharides(Vp2a-Ⅱ and Vp3) were isolated and identified from Apocynum venetum L. flowers, and their innate immune-stimulating functions and working mechanisms were evaluated in RAW264.7 cell...Two immunomodulatory polysaccharides(Vp2a-Ⅱ and Vp3) were isolated and identified from Apocynum venetum L. flowers, and their innate immune-stimulating functions and working mechanisms were evaluated in RAW264.7 cells. Both the level of released nitric oxide(NO) and expression of inducible nitric oxide synthase(iNOS) m RNA were significantly enhanced in the RAW264.7 macrophages cells treated by Vp2a-Ⅱ and Vp3. Vp2a-Ⅱ(100–800 μg/m L) and Vp3(400 μg/mL) could significantly increase the phagocytic activity of RAW264.7 cells and the secretion and m RNA expression of TNF-α and IL-6 in a concentrationdependent manner through affecting mitogen-activated protein kinase(MAPK) activity and nuclear factor κB(NF-κB) nuclear translocation. Vp2a-Ⅱ might activate the MAPK signaling pathways and induce the nuclear translocation of NF-κB p65, whilst Vp3 likely activated the NF-κB and MAPK signaling pathways without influencing the p38 MAPK route.展开更多
OBJECTIVE The emerging role of chronic inflammation is the major degenerative diseases of modern society such as periodontitis,atherosclerosis,rheumatoid arthritis,Parkinson′s disease and even cancer.Eight components...OBJECTIVE The emerging role of chronic inflammation is the major degenerative diseases of modern society such as periodontitis,atherosclerosis,rheumatoid arthritis,Parkinson′s disease and even cancer.Eight components were isolated from Derris laxiflora Benth.,In this study,we found these compounds from Derris laxiflora Benth suppress lipopolysaccharide-induced inflammatory response in murine macrophage(RAW 264.7)cells.METHODS RAW 264.7cells were cultured in DMEM media supplemented with 10%(V/V)heated-inactivated FBS,penicillin 100U·mL-1 and streptomycin 100μg·mL-1.The cells were incubated at 37℃in a humidified atmosphere of 5%CO2in air.RAW264.7cells were seeded in a 24-well plate at a density of 2×105 mL-1 and then incubated with or without LPS(100ng·mL-1)in the absence or presence of compounds for 24 h.Effects of these isolates on NO production were measured indirectly by analysis of nitrite levels using the Griess reaction.Quercetin was used as a positive control.RESULTS ight components were isolated from Derris laxiflora Benth.,including three new pterocarpans 7,6′-dihydroxy-3′-methoxypterocarpan(1),derrispisatin(2),derriscoumaronochromone(3)and three new flavonoids cis-3,4′-dihydroxy-5,7-dimethoxyflavan(4),derriflavanone B(5),iso-lupinenol(6)as well as two known ones,lonchocarpol A(7)and lonchocarpol D(8).The structures of these new compounds were determined by analysis of their spectroscopic data.Raw264.7 cells were treated with the compounds from Derris laxiflora Benth for 24 h.Among them,compounds 5,7 and 8 significantly suppressed the NO production in LPS-treated RAW264.7 cells with IC50 values<10μg·mL-1.CONCLUSION In this study,we found that compounds from Derris laxiflora Benth suppresses lipopolysaccharide-induced inflammatory response in murine Raw264.7 cells.展开更多
Macrophages play an important role in peripheral nerve regeneration,but the specific mechanism of regeneration is still unclear.Our preliminary findings indicated that neutrophil peptide 1 is an innate immune peptide ...Macrophages play an important role in peripheral nerve regeneration,but the specific mechanism of regeneration is still unclear.Our preliminary findings indicated that neutrophil peptide 1 is an innate immune peptide closely involved in peripheral nerve regeneration.However,the mechanism by which neutrophil peptide 1 enhances nerve regeneration remains unclear.This study was designed to investigate the relationship between neutrophil peptide 1 and macrophages in vivo and in vitro in peripheral nerve crush injury.The functions of RAW 264.7 cells we re elucidated by Cell Counting Kit-8 assay,flow cytometry,migration assays,phagocytosis assays,immunohistochemistry and enzyme-linked immunosorbent assay.Axonal debris phagocytosis was observed using the CUBIC(Clear,Unobstructed Brain/Body Imaging Cocktails and Computational analysis)optical clearing technique during Wallerian degeneration.Macrophage inflammatory factor expression in different polarization states was detected using a protein chip.The results showed that neutrophil peptide 1 promoted the prolife ration,migration and phagocytosis of macrophages,and CD206 expression on the surfa ce of macrophages,indicating M2 polarization.The axonal debris clearance rate during Wallerian degeneration was enhanced after neutrophil peptide 1 intervention.Neutrophil peptide 1 also downregulated inflammatory factors interleukin-1α,-6,-12,and tumor necrosis factor-αin invo and in vitro.Thus,the results suggest that neutrophil peptide 1 activates macrophages and accelerates Wallerian degeneration,which may be one mechanism by which neutrophil peptide 1 enhances peripheral nerve regeneration.展开更多
Objective: The effect of Chuanzhi Fang (ZGC) on the whole genome expression profile of RAW264.7 cells activated by lipopolysaccharide (LPS) was analyzed, and to explore the possible mechanism of action and core target...Objective: The effect of Chuanzhi Fang (ZGC) on the whole genome expression profile of RAW264.7 cells activated by lipopolysaccharide (LPS) was analyzed, and to explore the possible mechanism of action and core target of this formula on macrophage inflammatory injury at the overall level. Methods: A model of LPS-induced inflammation in RAW264.7 cells was constructed, and the effect of ZGC intervention on the genome-wide expression of inflammatory macrophages 3was examined by gene microarray technology, GO/KEGG enrichment analysis was performed for significantly differentially expressed genes among each group. Results: The results of genome-wide expression profiling microarray analysis showed that the ZGC intervention group upregulated the expression of 5 genes including C4bp and inhibited the expression of 22 genes including Mgat3, Psma6, and Siglecg relative to the LPS model group. KEGG signaling pathway analysis results showed that ZGC mainly acted through cytokine receptor interaction and the C-type lectin receptor signaling pathway. Conclusion: ZGC can interfere with the abnormal expression of 27 genes in inflammatory macrophages, and the related genes may exert corresponding anti-inflammatory effects by affecting cytokine receptor interactions, C-type lectin receptor signaling pathway, and TLR4/ NF-κB signaling pathway.展开更多
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
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
This study was designed to elucidate the immunoregulation of Ganoderma lucidum.HPLC fingerprint and spectrum-effect relationship of G.lucidum were established to predict the active compounds and BP neural network mode...This study was designed to elucidate the immunoregulation of Ganoderma lucidum.HPLC fingerprint and spectrum-effect relationship of G.lucidum were established to predict the active compounds and BP neural network model was established to predict the efficacy.Then the target compounds were identified by high resolution mass spectrometry.The results indicated that there are both enhanced immunity and immunosuppressive components in G.lucidum.BP neural network was trained with the common peak area and immune effi cacy index of G.lucidum fi ngerprint as samples,and a combined evaluation system of G.lucidum fi ngerprint effi cacy was established.The correlation coeffi cient R of BP network model was 0.98643,and the error of pharmacodynamic prediction results was in the ideal range.Eight compounds were identifi ed by high resolution mass spectrometry.The compounds related to immune activity in G.lucidum were determined in this study.展开更多
[Objectives]To explore the anti-inflammatory effect and mechanisms of different polar parts of Argyreia acuta Lour.[Methods]The effect of different polar parts of A.acuta Lour.on the viability of mouse RAW264.7 cells ...[Objectives]To explore the anti-inflammatory effect and mechanisms of different polar parts of Argyreia acuta Lour.[Methods]The effect of different polar parts of A.acuta Lour.on the viability of mouse RAW264.7 cells was detected by the CCK-8 method.Lipopolysaccharide(LPS)was used to stimulate RAW264.7 cells to establish an in vitro inflammation model,and the NO,TNF-αand IL-6 contents were determined by Griess method and ELISA assays,respectively.[Results]Within the range of 12.5-200μg/mL,all polar parts of A.acuta Lour.showed a proliferation effect on RAW264.7 cells,without cytotoxicity.The total extract,petroleum ether extract,ethyl acetate extract,and n-butanol extract all could inhibit the secretion of NO;and the petroleum ether extract and ethyl acetate extract could inhibit the secretion of TNF-αand IL-6.[Conclusions]The petroleum ether extract and ethyl acetate extract of A.acuta Lour.have significant anti-inflammatory activity,and the mechanism of action may be related to inhibiting the release of TNF-αand IL-6.展开更多
We reported the discovery of six novel coumarins, toddasirins A–F ( 1 – 6 ), each endowed with modified isoprenyl or geranyl side chains, derived from the roots of Toddalia asiatica. Comprehensive structural elucida...We reported the discovery of six novel coumarins, toddasirins A–F ( 1 – 6 ), each endowed with modified isoprenyl or geranyl side chains, derived from the roots of Toddalia asiatica. Comprehensive structural elucidation was achieved through multispectroscopic analyses, single-crystal X-ray diffraction experiments, and advanced quantum mechanical electronic circular dichroism (ECD) calculations. Furthermore, the anti-inflammatory activity of these compounds was assessed. Notably, compounds 1 – 3 and 6 demonstrated notable inhibitory effects on nitric oxide (NO) production in lipopolysaccharide (LPS)-induced RAW 264.7 cells, with 50% inhibitory concentration (IC50) values of 3.22, 4.78, 8.90, and 4.31 μmol·L^(−1), respectively.展开更多
基金supported by a National Research Foundation of Korea (NRF) grant funded by the Korea government (MSITNo. NRF-2020R1A2C1008527)
文摘Objective:To demonstrate the effect of dieckol from Eisenia bicyclis on osteoclastogenesis using RAW 264.7 cells.Methods:Murine macrophage RAW 264.7 cells were subjected to dieckol treatment,followed by treatment with receptor activator of nuclear factor kappa-B ligand(RANKL)to induce osteoclastogenesis.Tartrate-resistant acid phosphatase(TRAP)activity was examined using a TRAP activity kit.Western blotting analysis was conducted to examine the level of osteoclast-related factors,including TRAP and calcitonin receptor(CTR),transcriptional factors,including c-Fos,c-Jun,and nuclear factor of activated T cells cytoplasmic 1(NFATc1),nuclear factor kappa-B(NF-κB),extracellular signal-regulated kinase(ERK),and c-Jun N-terminal kinase(JNK).Immunofluorescence staining was conducted to examine the expression of c-Fos,c-Jun,and NFATc1.Results:Among the four phlorotannin compounds present in Eisenia bicyclis,dieckol significantly hindered osteoclast differentiation and expression of RANKL-induced TRAP and CTR.In addition,dieckol downregulated the expression levels of c-Fos,c-Jun,NFATc1,ERK,and JNK,and suppressed NF-κB signaling.Conclusions:Dieckol can suppress RANKL-induced osteoclastogenesis.Therefore,it has therapeutic potential in treating osteoclastogenesis-associated diseases.
文摘Curcumfa phaeocaulis Valeton is used in traditional Chinese medicines for the treatment of blood-related disorders such as blood stasis and inflammation.Our previous studies have found that sesquiterpenes were mainly isolated from C.wenyujin and C.phaeocaulis,while diarylheptanoids were the major compounds from C.kwangsiensis.And most of the isolated compounds exhibited remarkable
基金Supported by the Korea Basic Science Institute(No.C38915 and C030360)。
文摘Objective:To demonstrate the anti-inflammatory activity of Brassica napus L.hydrosols(BNH)in lipopolysaccharide(LPS)-stimulated RAW 264.7 cells.Methods:Composition analysis of BNH was conducted via gas chromatography-mass spectrometry after BNH were extracted.The nitric oxide(NO)production was measured using the Griess assay.Prostaglandin E2(PGE2)production was evaluated with enzyme-linked immunosorbent assay.The effects of BNH on LPS-induced pro-inflammatory enzymes including inducible nitric oxide synthase(iNOS)and cyclooxygenase-2(COX-2)were evaluated using Western blot analysis.Furthermore,phosphorylation of nuclear factor-kappa B(N F-k B)and nuclear translocation of N F-k B p65 were evaluated with Western blot analysis and immunofluorescence staining,respectively.Results:Compared with LPS-stimulated cells,BNH markedly decreased the generation of NO and PGE_(2)in LPS-stimulated RAW 264.7 cells(P<0.01 or P<0.05).Moreover,BNH inhibited protein levels of iNOS and COX-2(P<0.01).Phosphorylation of NF-k B and nuclear translocation of NF-k B p65 was significantly inhibited by BNH(P<0.01 or P<0.05).Conclusion:The anti-inflammatory activities of BNH were mediated via blockage of the N F-k B signaling pathways in LPS-stimulated RAW 264.7 cells.
基金supported by Research on Precision Nutrition and Health Food,Department of Science and Technology of Henan Province(CXJD2021006)。
文摘Two immunomodulatory polysaccharides(Vp2a-Ⅱ and Vp3) were isolated and identified from Apocynum venetum L. flowers, and their innate immune-stimulating functions and working mechanisms were evaluated in RAW264.7 cells. Both the level of released nitric oxide(NO) and expression of inducible nitric oxide synthase(iNOS) m RNA were significantly enhanced in the RAW264.7 macrophages cells treated by Vp2a-Ⅱ and Vp3. Vp2a-Ⅱ(100–800 μg/m L) and Vp3(400 μg/mL) could significantly increase the phagocytic activity of RAW264.7 cells and the secretion and m RNA expression of TNF-α and IL-6 in a concentrationdependent manner through affecting mitogen-activated protein kinase(MAPK) activity and nuclear factor κB(NF-κB) nuclear translocation. Vp2a-Ⅱ might activate the MAPK signaling pathways and induce the nuclear translocation of NF-κB p65, whilst Vp3 likely activated the NF-κB and MAPK signaling pathways without influencing the p38 MAPK route.
基金The project supported by Department of Industrial Technology,Ministry of Economic Affairs,Chinese TaipeiMedical and Pharmaceutical Industry Technology and Development Center
文摘OBJECTIVE The emerging role of chronic inflammation is the major degenerative diseases of modern society such as periodontitis,atherosclerosis,rheumatoid arthritis,Parkinson′s disease and even cancer.Eight components were isolated from Derris laxiflora Benth.,In this study,we found these compounds from Derris laxiflora Benth suppress lipopolysaccharide-induced inflammatory response in murine macrophage(RAW 264.7)cells.METHODS RAW 264.7cells were cultured in DMEM media supplemented with 10%(V/V)heated-inactivated FBS,penicillin 100U·mL-1 and streptomycin 100μg·mL-1.The cells were incubated at 37℃in a humidified atmosphere of 5%CO2in air.RAW264.7cells were seeded in a 24-well plate at a density of 2×105 mL-1 and then incubated with or without LPS(100ng·mL-1)in the absence or presence of compounds for 24 h.Effects of these isolates on NO production were measured indirectly by analysis of nitrite levels using the Griess reaction.Quercetin was used as a positive control.RESULTS ight components were isolated from Derris laxiflora Benth.,including three new pterocarpans 7,6′-dihydroxy-3′-methoxypterocarpan(1),derrispisatin(2),derriscoumaronochromone(3)and three new flavonoids cis-3,4′-dihydroxy-5,7-dimethoxyflavan(4),derriflavanone B(5),iso-lupinenol(6)as well as two known ones,lonchocarpol A(7)and lonchocarpol D(8).The structures of these new compounds were determined by analysis of their spectroscopic data.Raw264.7 cells were treated with the compounds from Derris laxiflora Benth for 24 h.Among them,compounds 5,7 and 8 significantly suppressed the NO production in LPS-treated RAW264.7 cells with IC50 values<10μg·mL-1.CONCLUSION In this study,we found that compounds from Derris laxiflora Benth suppresses lipopolysaccharide-induced inflammatory response in murine Raw264.7 cells.
基金supported by the National Natural Science Foundation of China,No.32371048(to YK)the Peking University People’s Hospital Research and Development Funds,No.RDX2021-01(to YK)the Natural Science Foundation of Beijing,No.7222198(to NH)。
文摘Macrophages play an important role in peripheral nerve regeneration,but the specific mechanism of regeneration is still unclear.Our preliminary findings indicated that neutrophil peptide 1 is an innate immune peptide closely involved in peripheral nerve regeneration.However,the mechanism by which neutrophil peptide 1 enhances nerve regeneration remains unclear.This study was designed to investigate the relationship between neutrophil peptide 1 and macrophages in vivo and in vitro in peripheral nerve crush injury.The functions of RAW 264.7 cells we re elucidated by Cell Counting Kit-8 assay,flow cytometry,migration assays,phagocytosis assays,immunohistochemistry and enzyme-linked immunosorbent assay.Axonal debris phagocytosis was observed using the CUBIC(Clear,Unobstructed Brain/Body Imaging Cocktails and Computational analysis)optical clearing technique during Wallerian degeneration.Macrophage inflammatory factor expression in different polarization states was detected using a protein chip.The results showed that neutrophil peptide 1 promoted the prolife ration,migration and phagocytosis of macrophages,and CD206 expression on the surfa ce of macrophages,indicating M2 polarization.The axonal debris clearance rate during Wallerian degeneration was enhanced after neutrophil peptide 1 intervention.Neutrophil peptide 1 also downregulated inflammatory factors interleukin-1α,-6,-12,and tumor necrosis factor-αin invo and in vitro.Thus,the results suggest that neutrophil peptide 1 activates macrophages and accelerates Wallerian degeneration,which may be one mechanism by which neutrophil peptide 1 enhances peripheral nerve regeneration.
基金Chinese Academy of Traditional Chinese Medicine Autonomous Topic Selection Project(No.ZZ2018017)Research Development Fund Project of the Medical Experimental Center of the Chinese Academy of Traditional Chinese Medicine(No.FZ2023003)。
文摘Objective: The effect of Chuanzhi Fang (ZGC) on the whole genome expression profile of RAW264.7 cells activated by lipopolysaccharide (LPS) was analyzed, and to explore the possible mechanism of action and core target of this formula on macrophage inflammatory injury at the overall level. Methods: A model of LPS-induced inflammation in RAW264.7 cells was constructed, and the effect of ZGC intervention on the genome-wide expression of inflammatory macrophages 3was examined by gene microarray technology, GO/KEGG enrichment analysis was performed for significantly differentially expressed genes among each group. Results: The results of genome-wide expression profiling microarray analysis showed that the ZGC intervention group upregulated the expression of 5 genes including C4bp and inhibited the expression of 22 genes including Mgat3, Psma6, and Siglecg relative to the LPS model group. KEGG signaling pathway analysis results showed that ZGC mainly acted through cytokine receptor interaction and the C-type lectin receptor signaling pathway. Conclusion: ZGC can interfere with the abnormal expression of 27 genes in inflammatory macrophages, and the related genes may exert corresponding anti-inflammatory effects by affecting cytokine receptor interactions, C-type lectin receptor signaling pathway, and TLR4/ NF-κB signaling pathway.
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.
基金This work was funded by the National Key R&D Program of China(2018YFD0400200)Key Project in Science and Technology Agency of Kaifeng City(1906006)Major Public Welfare Projects in Henan Province(201300110200).
文摘This study was designed to elucidate the immunoregulation of Ganoderma lucidum.HPLC fingerprint and spectrum-effect relationship of G.lucidum were established to predict the active compounds and BP neural network model was established to predict the efficacy.Then the target compounds were identified by high resolution mass spectrometry.The results indicated that there are both enhanced immunity and immunosuppressive components in G.lucidum.BP neural network was trained with the common peak area and immune effi cacy index of G.lucidum fi ngerprint as samples,and a combined evaluation system of G.lucidum fi ngerprint effi cacy was established.The correlation coeffi cient R of BP network model was 0.98643,and the error of pharmacodynamic prediction results was in the ideal range.Eight compounds were identifi ed by high resolution mass spectrometry.The compounds related to immune activity in G.lucidum were determined in this study.
文摘[Objectives]To explore the anti-inflammatory effect and mechanisms of different polar parts of Argyreia acuta Lour.[Methods]The effect of different polar parts of A.acuta Lour.on the viability of mouse RAW264.7 cells was detected by the CCK-8 method.Lipopolysaccharide(LPS)was used to stimulate RAW264.7 cells to establish an in vitro inflammation model,and the NO,TNF-αand IL-6 contents were determined by Griess method and ELISA assays,respectively.[Results]Within the range of 12.5-200μg/mL,all polar parts of A.acuta Lour.showed a proliferation effect on RAW264.7 cells,without cytotoxicity.The total extract,petroleum ether extract,ethyl acetate extract,and n-butanol extract all could inhibit the secretion of NO;and the petroleum ether extract and ethyl acetate extract could inhibit the secretion of TNF-αand IL-6.[Conclusions]The petroleum ether extract and ethyl acetate extract of A.acuta Lour.have significant anti-inflammatory activity,and the mechanism of action may be related to inhibiting the release of TNF-αand IL-6.
基金supported financially by the National Natural Science Foundation of China(No.82104014 and 82003605)the Key Laboratory Project of Colleges and Universities in Guangdong Province(No.2019KSYS005)the Guangdong Province Science and Technology Plan International Cooperation Project(No.2020A0505100052)。
文摘We reported the discovery of six novel coumarins, toddasirins A–F ( 1 – 6 ), each endowed with modified isoprenyl or geranyl side chains, derived from the roots of Toddalia asiatica. Comprehensive structural elucidation was achieved through multispectroscopic analyses, single-crystal X-ray diffraction experiments, and advanced quantum mechanical electronic circular dichroism (ECD) calculations. Furthermore, the anti-inflammatory activity of these compounds was assessed. Notably, compounds 1 – 3 and 6 demonstrated notable inhibitory effects on nitric oxide (NO) production in lipopolysaccharide (LPS)-induced RAW 264.7 cells, with 50% inhibitory concentration (IC50) values of 3.22, 4.78, 8.90, and 4.31 μmol·L^(−1), respectively.
