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
Objective To investigate in vitro cytotoxicity and oxidative stress response induced by multiwalled carbon nanotubes （MWCNTs）. Methods Cultured macrophages （murine RAW264.7 cells） and alveolar epithelium cells typ...Objective To investigate in vitro cytotoxicity and oxidative stress response induced by multiwalled carbon nanotubes （MWCNTs）. Methods Cultured macrophages （murine RAW264.7 cells） and alveolar epithelium cells type II （human A549 lung cells） were exposed to the blank control, DNA salt control, and the MWCNTs suspensions at 2.5, 10, 25, and 100 ug/mL for 24 h. Each treatment was evaluated by cell viability, cytotoxicity and oxidative stress. Results Overall, both cell lines had similar patterns in response to the cytotoxicity and oxidative stress of MWCNTs. DNA salt treatment showed no change compared to the blank control. In both cell lines, significant changes at the doses of 25 and 100 ug/mL treatments were found in cell viabilities, cytotoxicity, and oxidative stress indexes. The reactive oxygen species （ROS） generation was also found to be significantly higher at the dose of 10 ug/mL treatment, whereas no change was seen in most of the indexes. The ROS generation in both cell lines went up in minutes, reached the climax within an hour and faded down after several hours. Conclusion Exposure to MWCNTs resulted in a dose-dependent cytotoxicity in cultured RAW264.7 cells and A549 cells, that was closely correlated to the increased oxidative stress.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。
基金supported partly by a grant from Shanghai Science and Technology Committee International Collaboration Program (Project No. 055207078)
文摘Objective To investigate in vitro cytotoxicity and oxidative stress response induced by multiwalled carbon nanotubes （MWCNTs）. Methods Cultured macrophages （murine RAW264.7 cells） and alveolar epithelium cells type II （human A549 lung cells） were exposed to the blank control, DNA salt control, and the MWCNTs suspensions at 2.5, 10, 25, and 100 ug/mL for 24 h. Each treatment was evaluated by cell viability, cytotoxicity and oxidative stress. Results Overall, both cell lines had similar patterns in response to the cytotoxicity and oxidative stress of MWCNTs. DNA salt treatment showed no change compared to the blank control. In both cell lines, significant changes at the doses of 25 and 100 ug/mL treatments were found in cell viabilities, cytotoxicity, and oxidative stress indexes. The reactive oxygen species （ROS） generation was also found to be significantly higher at the dose of 10 ug/mL treatment, whereas no change was seen in most of the indexes. The ROS generation in both cell lines went up in minutes, reached the climax within an hour and faded down after several hours. Conclusion Exposure to MWCNTs resulted in a dose-dependent cytotoxicity in cultured RAW264.7 cells and A549 cells, that was closely correlated to the increased oxidative stress.
