• BMB Reports
• /
• v.52 no.6
• /
• pp.409-414
• /
• 2019

Natural compounds isolated from medicinal herbs and plants have immense significance in maintaining bone health. Hydrolysable tannins have been shown to possess a variety of medicinal properties including antiviral, anticancer, and anti-osteoclastogenic activities. As a part of a study on the discovery of alternative agent against skeletal diseases, we isolated a hydrolysable tannin, 2-O-digalloyl-1,3,4,6-tetra-O-galloyl-${\beta}$-D-glucose (DTOGG), from Galla Rhois and examined the effect on osteoclast formation and function. We found that DTOGG significantly inhibited receptor activator of nuclear factor-${\kappa}B$ ligand (RANKL)-induced osteoclast differentiation by downregulating the expression of the key regulator in osteoclastogenesis as well as osteoclast-related genes. Analysis of RANKL/RANK signaling revealed that DTOGG impaired activation of $I{\kappa}B{\alpha}$ and p65 in the nuclear factor kappa-lightchain-enhancer of activated B cells (NF-${\kappa}B$) signaling pathway. Furthermore, DTOGG reduced bone resorbing activity of osteoclasts, compared to the vehicle-treated control. These results suggest that DTOGG could be a useful natural compound to manage osteoclast-mediated skeletal diseases.

• Journal of Korean Traditional Oncology
• /
• v.16 no.2
• /
• pp.25-34
• /
• 2011

Objectives : Citrus is the fruit that is readily available around us. Therefore, we investigated the anti-inflammatory effects of fraction isolated from the Citrus hassaku pericarp in RAW264.7 macrophage cells. Methods : The effects of fraction from Citrus hassaku pericarp on cell viability on RAW264.7 cells were measured by the MTT assay. The mRNA levels of iNOS and COX-2, its protein level by fraction of Citrus hassaku pericarp treatment in RAW264.7 macrophage cells were investigated by RT-PCR and immunoblots. Nitrite accumulation in the culture was measured colorimetrically by the Griess reaction using a Griess reagent. The amount of IL-6 and TNF-${\alpha}$ production was determined using an enzyme-linked immunosorbent assay (ELISA) kit. Results : The results indicated that the fraction of Citrus hassaku pericarp concentration highly suppressed lipopolysaccharide (LPS)-induced nitric oxide (NO) and IL-6 productions without a cytotoxic effect on RAW264.7 cells. fraction of Citrus hassaku pericarp inhibited the expressions of LPS-induced iNOS and COX-2 protein and their mRNA in a dose-dependent manner. Particularly, fraction of Citrus hassaku pericarp suppressed the level of nuclear factor-${\kappa}B$ (NF-${\kappa}B$) activity, which was linked with the suppression of LPS-induced phosphorylation of p65 at serine 276 and p65 translocation into nuclei, but not MAPK signaling. In addition, treatment with fraction of Citrus hassaku pericarp inhibited the production of IL-6 and TNF-${\alpha}$ in LPS-stimulated RAW264.7 cells. Conclusion : Our results indicate that fraction of Citrus hassaku pericarp potentially inhibits the biomarkers related to inflammation through the blocking of NF-${\kappa}B$ p65 activation, and it may be a potential therapeutic candidate for the treatment of inflammatory diseases.

• Korean Journal of Food Science and Technology
• /
• v.49 no.4
• /
• pp.453-458
• /
• 2017

This study compared the immuno-modulatory effects of ethanol extracts (A. muricata L. ethanol extracts, ALE) and crude polysaccharide fraction (A. muricata L. crude polysaccharide fraction, ALP) from Annona muricata L. in macrophages. Immuno-modulatory activity was determined by assessing cell viability, nitric oxide (NO) production, inducible NO synthase (iNOS) expression and cytokine production in RAW 264.7 a macrophage cell line. Both ALE and ALP treatment did not affect cytotoxicity, and ALP treatment significantly increased NO production. Additionally, cytokine production [tumor necrosis factor ($TNF-{\alpha}$; $2909.04{\pm}4.1pg/mL$), interleukin (IL)-6; $662.84{\pm}5.3pg/mL$, and $IL-1{\beta}$; $852.37{\pm}2.2pg/mL$), was highly increased in the ALP ($250{\mu}g/mL$) treated group compared to the ALE ($250{\mu}g/mL$) treated group ($TNF-{\alpha}$; $1564.50{\pm}6.1pg/mL$, IL-6; $517.24{\pm}4.1pg/mL$ and $IL-1{\beta}$; $237.23{\pm}1.8pg/mL$). Moreover, ALP treatment considerably increased the expression of mitogen-activated protein kinase (MAPKs) and nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$) in the macrophages. Therefore, ALP can induce macrophage activation through MAPK and $NF-{\kappa}B$ signaling and this can be a potential candidate for development of nutraceuticals.

• Korean Journal of Plant Resources
• /
• v.31 no.3
• /
• pp.211-217
• /
• 2018

Hibiscus syriacus (H. syriacus) as the national flower of Korea has been used as the herbal medicine in Asia. In this study, we evaluated the anti-inflammatory effect of 70% ethanol extracts from the root of Hibiscus syriacus (RHS-E70) and elucidated the potential signaling pathway in LPS-stimulated RAW264.7 cells. RHS-E70 dose-dependently suppressed NO production by inhibiting iNOS and IL-${\beta}$ expression in LPS-stimulated RAW264.7 cells. RHS-E70 inhibited the phosphorylation and degradation of $I{\kappa}B-{\alpha}$, which contributed to the inhibition of p65 nuclear accumulation and NF-${\kappa}B$ activation. Furthermore, RHS-E70 suppressed the phosphorylation of ERK1/2 and p38, which results in the inhibition of ATF2 phosphorylation and subsequent nuclear accumulation. These results indicate that RHS-E70 may exert anti-inflammatory activity by inhibiting NF-${\kappa}B$ and MAPK/ATF2 signaling. From these findings, RHS-E70 has potential to be a candidate for the development of chemopreventive or therapeutic agents for the inflammatory diseases.

• Journal of Applied Biological Chemistry
• /
• v.61 no.3
• /
• pp.275-281
• /
• 2018

Pinus densiflora root (PDR) is used as a medicinal plant. In this study, we investigated whether the PDR extract has anti-inflammatory activities. Cell viability assays showed that the extract was not toxic toward RAW 264.7 cells at concentrations up to $10{\mu}g/mL$. At $10{\mu}g/mL$, the extract decreased nitric oxide (NO) content to 40% of the control level. The protein expression of inducible nitric oxide synthase (iNOS), which generates NO, decreased with increasing concentrations of the extract. Prostaglandin $E_2$ ($PGE_2$) levels were significantly inhibited by over 50% in the presence of $10{\mu}g/mL$ of the extract. The protein expression of cyclooxygenase-2 (COX-2), which generates $PGE_2$, decreased with increasing concentrations of the extract. Proinflammatory cytokines, such as tumor necrosis factor-alpha ($TNF-{\alpha}$), interleukin-6 (IL-6), and $IL-1{\beta}$, were detected in RAW 264.7 cells after lipopolysaccharide (LPS) treatment. The extract did not affect the levels of $TNF-{\alpha}$ and IL-6, but it significantly inhibited the level of $IL-1{\beta}$. It also completely inhibited the transcription of nuclear factor-kappaB ($NF-{\kappa}B$). These results indicate that the PDR extract reduces inflammatory response-related proteins, such as NO, $PGE_2$, iNOS, and COX-2, in LPS-induced RAW 264.7 cells via the regulation of $NF-{\kappa}B$. Consequently, we have provided a mechanism to explain the anti-inflammatory effect of the PDR extract; that is, it exerts such an effect by regulating $NF-{\kappa}B$. The PDR extract can therefore be considered as an effective anti-inflammatory agent.

• Journal of Pharmacopuncture
• /
• v.6 no.2
• /
• pp.45-56
• /
• 2003

Objective : The purpose of this study was to investigate the effects of Bee Venom on the lipopolysaccharide(LPS), sodium nitroprusside(SNP), hydrogen peroxide$(H_2O_2)$-induced expression inducilble nitric oxide synthetase(iNOS), tumor necrosis factor-${\alpha}$(TNF-${\alpha}$) and nuclear factor kappa B(NF-kB) in RAW 264.7 cells, a murine macrophage cell line. Method : The expressions of expression iNOS and TNF-${\alpha}$ were determined by western blotting with corresponding antibodies. The expressions of expression NF-kB was assayed by EMSA method. Results : 1. The 0.5, 1 and $5{\mu}g/mg$ of bee venom on LPS-induced expression of iNOS, the $5{\mu}g/mg$ of bee venom on SNP-induced expression of iNOS and the $1{\mu}g/mg$ of bee venom on $H_2O_2$-induced expression of iNOS compared with control were inhibited significantly. 2. The 0.5, 1 and $5{\mu}g/mg$ of bee venom inhibited significantly LPS and $H_2O_2$-induced expression of TNF-${\alpha}$ compared with control, respectively. The $0.5{\mu}g/mg$ of bee venom increased significantly SNP-induced expression of TNF-${\alpha}$ compared with control. 3. The $5{\mu}g/mg$ of bee venom on LPS-induced expression of NF-kB, the $0.5{\mu}g/mg$ of bee venom on SNP-induced expression of NF-kB and the 0.5, $5{\mu}g/mg$ of bee venom on $H_2O_2$-induced expression of NF-kB were inhibited significantly compared with control, respectively.

• Journal of Acupuncture Research
• /
• v.29 no.1
• /
• pp.75-87
• /
• 2012

Objectives : The purpose of this study is to inquire into the effect of different concentrations of bee venom pharmacopuncture to inhibit genesis of pro-inflammatory cytokines and to inhibit nuclear factor (NF)-${\kappa}B$ activation on type II collagen induced arthritis. Methods : The experiment was divided into category of the normal group (NOR)-no treated group, control group (CON)-CIA (collagen induced arthritis) induced group, and 4,000 : 1 bee venom group (BV-L)- 4000:1 bee venom pharmacopuncture treated group after CIA, and 2000:1 bee venom group (BV-H)- 2,000 : 1 Bee venom pharmacopuncture treated group after CIA. RA was induced in the mice via injecting $50{\mu}{\ell}$ C II mixed CFA. The bee venom pharmacopuncture was applied on $ST_{35}$ for 19 days from the 3rd day of RA inducement. To research the effect on the expression of IKK ($I{\kappa}B$ kinase), iNOS (inducible nitric oxide synthase) & COX-2 (cyclooxygenase-2) mRNA, RT-PCR was performed on synovial membrane cells from the knee joint of CIA mice. Results : The PMA-induced $I{\kappa}B$ kinase (IKK), inducible nitric oxide synthase (iNOS) and cyclooxygenase -2 (COX-2) mRNA expression were dose-dependantly decreased in bee venom treated with synoviocytes. In mice treated with bee venom pharmacopuncture, foot thickness and the damage of synovial membranes of the joint was lessened, and the activation of RA-related pro-inflammatory cytokines such as MIF, TNF-${\alpha}$ and MMP-9 was significantly decreased. The activation of iNOS and COX-2 was suppressed by the inhibition of NF-${\kappa}B$. In addition, each data was shown that 2,000 : 1 bee venom pharmacopuncture was more effective than 4,000 : 1 bee venom pharmacopuncture. Conclusions : It is speculated that bee venom pharmacopuncture has the therapeutic effect of palliating the damage of the synovial membrane and inflammation on RA by suppressing of NF-${\kappa}B$ activation.

• Proceedings of the Plant Resources Society of Korea Conference
• /
• 2019.10a
• /
• pp.67-67
• /
• 2019

Vaccinium oldhamii (V. oldhamii) has been reported to exert a variety of the pharmacological properties such as anti-oxidant activity, anti-cancer activity, and inhibitory activity of ${\alpha}$-amylase and acetylcholinesterase. However, the anti-inflammatory activity of V. oldhamii has not been studied. In this study, we aimed to investigate anti-inflammatory activity of the stem extracts from V. oldhamii, and to elucidate the potential mechanisms in LPS-stimulated RAW264.7 cells. Among VOS, VOL and VOF, the inhibitory effect of NO and PGE2 production induced by LPS was highest in VOS treatment. Thus, VOS was selected for the further study. VOS dose-dependently blocked LPS-induced NO and PGE2 production by inhibiting iNOS and COX-2 expression, respectively. VOS inhibited the expression of pro-inflammatory cytokines such as $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$. In addition, VOS suppressed TRAP activity and attenuated the expression of the osteoclast-specific genes such as NFATc1, c-FOS, TRAP, MMP-9, cathepsin K, CA2, OSCAR and ATPv06d2. VOS inhibited LPS-induced $NF-{\kappa}B$ signaling activation through blocking $I{\kappa}B-{\alpha}$ degradation and p65 nuclear accumulation. VOS inhibited MAPK signaling activation by attenuating the phosphorylation of ERK1/2, p38 and JNK. Furthermore, VOS inhibited ATF2 phosphorylation and blocked ATF2 nuclear accumulation. From these findings, VOS has potential to be a candidate for the development of chemopreventive or therapeutic agents for the inflammatory diseases.

• International Journal of Oral Biology
• /
• v.37 no.3
• /
• pp.137-145
• /
• 2012

Aggregatibacter actinomycetemcomitans is the most important etiologic agent of aggressive periodontitis and can interact with endothelial cells. Monocyte chemoattractant protein-1 (MCP-1) and interleukin-8 (IL-8) are chemokines, playing important roles in periodontal pathogenesis. In our current study, the effects of A. actinomycetemcomitans on the production of MCP-1 and IL-8 by human umbilical vein endothelial cells (HUVEC) were investigated. A. actinomycetemcomitans strongly induced the gene expression and protein release of both MCP-1 and IL-8 in a dose- and time-dependent manner. Dead A. actinomycetemcomitans cells were as effective as live bacteria in this induction. Treatment of HUVEC with cytochalasin D, an inhibitor of endocytosis, did not affect the mRNA up-regulation of MCP-1 and IL-8 by A. actinomycetemcomitans. However, genistein, an inhibitor of protein tyrosine kinases, substantially inhibited the MCP-1 and IL-8 production by A. actinomycetemcomitans, whereas pharmacological inhibition of each of three members of mitogen-activated protein (MAP) kinase family had little effect. Furthermore, gel shift assays showed that A. actinomycetemcomitans induces a biphasic activation (early at 1-2 h and late at 8-16 h) of nuclear factor-${\kappa}B$ (NF-${\kappa}B$) and an early brief activation (0.5-2 h) of activator protein-1 (AP-1). Activation of canonical NF-${\kappa}B$ pathway ($I{\kappa}B$ kinase activation and $I{\kappa}B-{\alpha}$ degradation) was also demonstrated in these experiments. Although lipopolysaccharide from A. actinomycetemcomitans also induced NF-${\kappa}B$ activation, this activation profile over time differed from that of live A. actinomycetemcomitans. These results suggest that the expression of MCP-1 and IL-8 is potently increased by A. actinomycetemcomitans in endothelial cells, and that the viability of A. actinomycetemcomitans and bacterial internalization are not required for this effect, whereas the activation of protein tyrosine kinase(s), NF-${\kappa}B$, and AP-1 appears to play important roles. The secretion of high levels of MCP-1 and IL-8 resulting from interactions of A. actinomycetemcomitans with endothelial cells may thus contribute to the pathogenesis of aggressive periodontitis.

• Korean Journal of Medicinal Crop Science
• /
• v.19 no.4
• /
• pp.217-226
• /
• 2011

The study was conducted to investigate candidate materials as anti-inflammation agent from plant resources. Activities of 33 plant parts extracts with the final concentration of 5${\mu}g/ml$ were evaluated on the several inflammation-related markers such as the release of proinflammatoty cytokine [tumor necrosis factor-alpha (TNF-${\alpha}$) & interleukin-6 (IL-6)], nitric oxide (NO), the expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) and inhibitor of nuclear factor kappa-B alpha ($I{\kappa}-B{\alpha}$) in lipopolysaccharide (LPS)-treated RAW 264.7 cells. The extracts in the final concentration of 10 ${\mu}g/ml$ were also screened on peroxynitrite (ONOO$^-$) scavenging activity. Eleven extracts selected from the screening assay were verified on the inhibition activity on peroxynitrite and total reactive species oxygen (ROS) in the several concentrations. As results, Alpinia officinarum Hance (rhizome), Inula britannica var. chinensis Regel (flower), Ulmus arvifolia Jacq (trunk peel) and Aster scaber Thunb. (aerial part) showed comparatively potent anti-inflammatory activities in vitro cells or chemical level systems, and then these four plant parts should be studied on the antiinflammatory mechanism by further studies.