In the present study, 3-methylcarbazole and 1-methoxy-3-methylcarbazole were isolated from the culture of Streptomyces sp. LJK109, endophyte of Alpinia galanga Swartz. 3-methylcarbazole, a carbazole derivative, has be...In the present study, 3-methylcarbazole and 1-methoxy-3-methylcarbazole were isolated from the culture of Streptomyces sp. LJK109, endophyte of Alpinia galanga Swartz. 3-methylcarbazole, a carbazole derivative, has been found to be highly potent as anti-inflammatory agent. The immunomodulatory activity of these agents in toll like receptor (TLR)-activated RAW 264.7 macrophages induced by lipopolysaccharide (LPS), Poly(I:C), and pam3CSK was investigated by assessing nitric oxide (NO) and pro-inflammatory cytokines. The 3-methylcarbazoles dose-dependently suppressed the release of NO, PGE2, TNF-α, IL-1β, IL-6 and IL-10 in LPS- and pam3CSK-activated macrophages but not in Poly(I:C)-activated macrophages. Our results suggest that 3-methylcarbazoles can be further developed as a promising anti-inflammatory remedy.展开更多
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
Geldanamycin (1) had been isolated as a major compound from Streptomyces zerumbet W14;an endophyte of Zingiber zerumbet (L.) Smith. Two new geldanamycin derivatives;17-(tryptamine)-17-demethoxygeldanamycin (2) and 17...Geldanamycin (1) had been isolated as a major compound from Streptomyces zerumbet W14;an endophyte of Zingiber zerumbet (L.) Smith. Two new geldanamycin derivatives;17-(tryptamine)-17-demethoxygeldanamycin (2) and 17-(5’-methoxytryptamine)-17-demethoxygeldanamycin (3) were synthe- sized and their anti-inflammatory activity was evaluated in LPS-induced macrophage RAW 264.7 cells by investigating their effects on the inhibition of production of NO, PGE2, TNF-α, IL-1β, IL-6 and IL-10. The data obtained were consistent with the modulation of TNF-α, IL-1β, IL-6, IL-10 production by these derivatives at concentration of 1 to 5 μg/ml. A similar effect was also observed when LPS-induced NO release and PGE2 production were tested. The inhibitory effects were shown in concentration-dependent manners. From the obtained results, it was concluded that two new gelda- namycin derivatives possess anti-inflammatory activity on LPS-induced RAW 264.7 cells. They could be useful for the management of inflammatory diseases.展开更多
We previously demonstrated that 2-hydroxypropyltrimethyl ammonium chloride chitosan(HACC)promoted the production of nitric oxide(NO)and proinflammatory cytokines by activating the mitogen-activated protein kinases(MAP...We previously demonstrated that 2-hydroxypropyltrimethyl ammonium chloride chitosan(HACC)promoted the production of nitric oxide(NO)and proinflammatory cytokines by activating the mitogen-activated protein kinases(MAPK)and Janus kinase(JAK)/STAT pathways in RAW 264.7 cells,indicating good immunomodulatory activity of HACC.In this study,to further investigate the immunomodulatory mechanisms of HACC,we determined the roles of phosphatidylinositol 3-kinase(PI3K)/Akt,activating protein(AP-1)and nuclear factor kappa B(NF-κB)in HACC-induced activation of RAW 264.7 cells by the western blotting.The results suggest that HACC promoted the phosphorylation of p85 and Akt.Furthermore,c-Jun and p65 were also increased after the treatment of RAW 264.7 cells with HACC,indicating the translocation of NF-κB and AP-1 from cytoplasm to nucleus.In addition,as scanning electron microscopy(SEM)analysis shows,the cell morphology changed after HACC treatment.These findings indicate that HACC activated MAPK,JAK/STAT,and PI3K/Akt signaling pathways dependent on AP-1 and NF-κB activation in RAW 264.7 cells,ultimately leading to the increase of NO and cytokines.展开更多
背景:前期研究表明葛根素干预后破骨细胞的分化被抑制,Notch1、HES1、Jagged1等Notch信号通路相关蛋白表达量下降,但Notch1信号通路对于葛根素抑制破骨细胞分化的作用机制尚不明确。目的:探究Notch信号通路对葛根素抑制小鼠巨噬细胞Raw2...背景:前期研究表明葛根素干预后破骨细胞的分化被抑制,Notch1、HES1、Jagged1等Notch信号通路相关蛋白表达量下降,但Notch1信号通路对于葛根素抑制破骨细胞分化的作用机制尚不明确。目的:探究Notch信号通路对葛根素抑制小鼠巨噬细胞Raw264.7分化为破骨细胞的影响。方法:将Raw264.7细胞分为7组干预培养,空白对照组采用DMEM高糖完全培养基培养,破骨细胞诱导组采用破骨诱导培养基培养,葛根素干预组在破骨诱导的同时加入50μmol/L葛根素培养,葛根素+Notch1 si RNA对照组、葛根素+Notch1 si RNA组、葛根素+Notch1过表达对照组、葛根素+Notch1过表达组分别采用Notch1 si RNA对照序列、Notch1 si RNA序列、Notch1过表达对照质粒、Notch1过表达质粒转染Raw264.7细胞后,加入破骨诱导培养基和葛根素进行培养。培养7 d后,采用抗酒石酸酸性磷酸酶染色观察破骨细胞的数量和大小,F-actin染色观察破骨细胞骨架形成情况,RT-PCR检测破骨细胞形成标志物的基因表达水平。结果与结论:(1)抗酒石酸酸性磷酸酶染色显示:葛根素干预可以抑制破骨细胞的生成,Notch1沉默会进一步减少破骨细胞的生成数量,Notch1过表达后破骨细胞生成数量明显增加;(2)F-actin染色显示:Raw264.7细胞经破骨诱导可以形成边界清晰的F-actin环,葛根素干预会抑制细胞骨架的形成,Notch1沉默会增强葛根素的抑制作用,而Notch1过表达则能减弱葛根素的抑制作用;(3)RT-PCR检测显示,葛根素可以抑制抗酒石酸酸性磷酸酶、组织蛋白酶K和c-Fos的m RNA表达,Notch1沉默后上述3个因子的m RNA表达进一步降低,Notch1过表达后上述3个因子的m RNA表达增加。结果表明:Notch信号通路在Raw264.7细胞分化为破骨细胞的过程中发挥作用,葛根素通过抑制Notch信号通路抑制Raw264.7细胞分化为破骨细胞。展开更多
文摘In the present study, 3-methylcarbazole and 1-methoxy-3-methylcarbazole were isolated from the culture of Streptomyces sp. LJK109, endophyte of Alpinia galanga Swartz. 3-methylcarbazole, a carbazole derivative, has been found to be highly potent as anti-inflammatory agent. The immunomodulatory activity of these agents in toll like receptor (TLR)-activated RAW 264.7 macrophages induced by lipopolysaccharide (LPS), Poly(I:C), and pam3CSK was investigated by assessing nitric oxide (NO) and pro-inflammatory cytokines. The 3-methylcarbazoles dose-dependently suppressed the release of NO, PGE2, TNF-α, IL-1β, IL-6 and IL-10 in LPS- and pam3CSK-activated macrophages but not in Poly(I:C)-activated macrophages. Our results suggest that 3-methylcarbazoles can be further developed as a promising anti-inflammatory remedy.
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Geldanamycin (1) had been isolated as a major compound from Streptomyces zerumbet W14;an endophyte of Zingiber zerumbet (L.) Smith. Two new geldanamycin derivatives;17-(tryptamine)-17-demethoxygeldanamycin (2) and 17-(5’-methoxytryptamine)-17-demethoxygeldanamycin (3) were synthe- sized and their anti-inflammatory activity was evaluated in LPS-induced macrophage RAW 264.7 cells by investigating their effects on the inhibition of production of NO, PGE2, TNF-α, IL-1β, IL-6 and IL-10. The data obtained were consistent with the modulation of TNF-α, IL-1β, IL-6, IL-10 production by these derivatives at concentration of 1 to 5 μg/ml. A similar effect was also observed when LPS-induced NO release and PGE2 production were tested. The inhibitory effects were shown in concentration-dependent manners. From the obtained results, it was concluded that two new gelda- namycin derivatives possess anti-inflammatory activity on LPS-induced RAW 264.7 cells. They could be useful for the management of inflammatory diseases.
基金Supported by the National Key R&D Program of China(No.2018YFC0311305)the Key Research and Development Program of Shandong Province(Nos.2019GHY112015,2019YYSP028)。
文摘We previously demonstrated that 2-hydroxypropyltrimethyl ammonium chloride chitosan(HACC)promoted the production of nitric oxide(NO)and proinflammatory cytokines by activating the mitogen-activated protein kinases(MAPK)and Janus kinase(JAK)/STAT pathways in RAW 264.7 cells,indicating good immunomodulatory activity of HACC.In this study,to further investigate the immunomodulatory mechanisms of HACC,we determined the roles of phosphatidylinositol 3-kinase(PI3K)/Akt,activating protein(AP-1)and nuclear factor kappa B(NF-κB)in HACC-induced activation of RAW 264.7 cells by the western blotting.The results suggest that HACC promoted the phosphorylation of p85 and Akt.Furthermore,c-Jun and p65 were also increased after the treatment of RAW 264.7 cells with HACC,indicating the translocation of NF-κB and AP-1 from cytoplasm to nucleus.In addition,as scanning electron microscopy(SEM)analysis shows,the cell morphology changed after HACC treatment.These findings indicate that HACC activated MAPK,JAK/STAT,and PI3K/Akt signaling pathways dependent on AP-1 and NF-κB activation in RAW 264.7 cells,ultimately leading to the increase of NO and cytokines.
文摘背景:前期研究表明葛根素干预后破骨细胞的分化被抑制,Notch1、HES1、Jagged1等Notch信号通路相关蛋白表达量下降,但Notch1信号通路对于葛根素抑制破骨细胞分化的作用机制尚不明确。目的:探究Notch信号通路对葛根素抑制小鼠巨噬细胞Raw264.7分化为破骨细胞的影响。方法:将Raw264.7细胞分为7组干预培养,空白对照组采用DMEM高糖完全培养基培养,破骨细胞诱导组采用破骨诱导培养基培养,葛根素干预组在破骨诱导的同时加入50μmol/L葛根素培养,葛根素+Notch1 si RNA对照组、葛根素+Notch1 si RNA组、葛根素+Notch1过表达对照组、葛根素+Notch1过表达组分别采用Notch1 si RNA对照序列、Notch1 si RNA序列、Notch1过表达对照质粒、Notch1过表达质粒转染Raw264.7细胞后,加入破骨诱导培养基和葛根素进行培养。培养7 d后,采用抗酒石酸酸性磷酸酶染色观察破骨细胞的数量和大小,F-actin染色观察破骨细胞骨架形成情况,RT-PCR检测破骨细胞形成标志物的基因表达水平。结果与结论:(1)抗酒石酸酸性磷酸酶染色显示:葛根素干预可以抑制破骨细胞的生成,Notch1沉默会进一步减少破骨细胞的生成数量,Notch1过表达后破骨细胞生成数量明显增加;(2)F-actin染色显示:Raw264.7细胞经破骨诱导可以形成边界清晰的F-actin环,葛根素干预会抑制细胞骨架的形成,Notch1沉默会增强葛根素的抑制作用,而Notch1过表达则能减弱葛根素的抑制作用;(3)RT-PCR检测显示,葛根素可以抑制抗酒石酸酸性磷酸酶、组织蛋白酶K和c-Fos的m RNA表达,Notch1沉默后上述3个因子的m RNA表达进一步降低,Notch1过表达后上述3个因子的m RNA表达增加。结果表明:Notch信号通路在Raw264.7细胞分化为破骨细胞的过程中发挥作用,葛根素通过抑制Notch信号通路抑制Raw264.7细胞分化为破骨细胞。
