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Antiaging Activity of Mixed Extracts from Korean Medicinal Herbs on HS68 Skin Fibroblast (한약재복합 추출물의 인간피부섬유아세포 HS68에 대한 항노화 효과)

  • Shin, Dong-Chul;Kim, Gwui-Cheol;Song, Si-Young;Kim, Hee-Jin;Yang, Jae-Chan;Lee, Yong-Hwa;Kim, Bo-Ae
    • The Korea Journal of Herbology
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    • v.29 no.2
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    • pp.39-45
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    • 2014
  • Objectives : The purpose of this study was to investigate antiaging and antioxidant effects on cultured human skin fibroblast with 80% ethanol extracts of plants including of stem of Dendropanax morbifera, Corni fructus and Lycii Fructus. Methods : An ethanol extract of three medicinal plants including stem of Dendropanax morbifera, Corni fructus and Lycii Fructus. Extracts were assessed to determine the mechanism of antioxidant and antiaging activities. Antioxidant activity of extract was evaluated by two different assays as 2,2-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging and super oxide dismutase (SOD) like activities. These extracts were tested for cell viability on HS68 skin fibroblast by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. We investigated the effects of Ultraviolet-B irradiation on cytotoxicity, type 1 collagen, elastin level and oxidative damage in cultured human skin fibroblast (HS68). Recently, many studies have reported that elastin is also involved in inhibiting or repairing wrinkle formation, although collagen is a major factor in the skin wrinkle formation. Results : The extracts obtained dose-dependently increased the scavenging activity on DPPH radical scavenging activity and SOD like activity. The extracts of complex herbal medicine showed low cytotoxicity as more than 100% cell viability in 100ppm/ml concentration. HS68 fibroblasts were survived 70% at 120 $mJ/cm^2$ UVB irradiation and treated tumor necrosis factor (TNF)-alpha. The levels of aging factors and cytotoxicity were decreased by ethanol extract of complex herbal medicine. Conclusions : These results suggest that ethanol extracts of complex medicinal plants of including of stem of Dendropanax morbifera, Corni fructus and Lycii Fructus may have value as the potential antioxidant and antiaging medicinal plant.

Thuja orientalis leaves extract protects dopaminergic neurons against MPTP-induced neurotoxicity via inhibiting inflammatory action (MPTP로 유도된 Parkinson's disease 동물 모델에서 항염증효과를 통한 측백엽의 도파민신경보호 효과)

  • Park, Gunhyuk;Kim, Hyo Geun;Ju, Mi Sun;Kim, Ae-Jung;Oh, Myung Sook
    • The Korea Journal of Herbology
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    • v.29 no.3
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    • pp.27-33
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    • 2014
  • Objectives : The aim of this study was to investigate the protective effect of extract of Thuja orientalis leaves (TOFE) against 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced neurotoxicity by inhibition of inflammation in in vitro and in vivo models of Parkinson's disease (PD). Methods : We evaluated the effect of TOFE against lipopolysaccharide (LPS)/1-methyl-4-phenylpyridinium ($MPP^+$) toxicity using nitric oxide (NO) assay, inducible NO synthase and cyclooxygenase 2 western blot, tyrosine hydroxylase and microglia activation immunohistochemistry (IHC) in BV2 cell, primary rat mesencephalic neurons, or C57BL/6 mice. We also evaluated the effect of TOFE in mice PD model induced by MPTP. C57BL/6 mice were treated with TOFE 50 mg/kg for 5 days and were injected intraperitoneally with four administrations of MPTP on the last day. We conducted behavioral tests and IHC analysis to see how TOFE affect MPTP-induced neuronal loss of dopaminergic neurons in substantia nigra pars compacta (SNpc) and striatum (ST) of mice. To assess the anti-inflammation effects, we carried out glial fibrillary acidic protein and macrophage-1 antigen integrin alpha M in IHC in SNpc and ST of mice. Results : In an in vitro system, TOFE decreasesd NO generations in BV2 cells. TOFE protected dopaminergic cells against LPS or $MPP^+$-induced toxicity in primary mesencephalic dopaminergic neurons. In vivo system, TOFE at 50 mg/kg treated group showed improved motor deteriorations than the MPTP only treated group and TOFE significantly protected striatal dopaminergic damage from MPTP-induced neurotoxicity in mice. Moreover, TOFE inhibited activation of astrocyte and microglia in SNpc and ST of the mice. Conclusions : We concluded that TOFE showed anti-parkinsonian effect by protection of dopaminergic neurons against MPTP toxicity through anti-inflammatory actions.

Effect of Gami-Chunggisan on Antioxidant and Pro-Inflammatory Cytokine (가미청기산(加味淸肌散)이 항산화와 염증성 사이토카인에 미치는 영향)

  • Lee, Youn-Jeong;Sim, Boo-Yong;Lee, Hae-Jin;Bak, Ji-Won;Kim, Dong-Hee
    • The Korea Journal of Herbology
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    • v.29 no.4
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    • pp.69-76
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    • 2014
  • Objectives : Gami-Chunggisan extract (GCE) is one of the oriental traditional medicine. We investigated the antioxidant effect and reduction of pro-inflammatory cytokine as a functional ingredient for cosmetic products from the GCE. Methods : GCE was prepared by extracting with 80% ethanol. We analyzed total polyphenol and antioxidant activities. To evaluate antioxidant activity, we measured 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 1,1-diphenyl-2-picryl-hydrazyl (DPPH) free radical scavenging assay. Also we measured the production of reactive oxygen species (ROS) and nitric oxide (NO) on Raw264.7 cells. We researched reduction of anti-inflammatory cytokines from concentration of GCE on Raw264.7 cells. Results : Total polyphenol quantity of GCE was included 46.6 mg/g. The GCE showed ABTS free radical scavenging ability with more than 89% at $1000{\mu}g/m{\ell}$. In addition the DPPH free radical scavenging ability from the GCE was activated over 93% at $1000{\mu}g/m{\ell}$. Production of the ROS was decreased by approximately 26%, upon the GCE treatment at concentration of $100{\mu}g/m{\ell}$. The GCE at $100{\mu}g/m{\ell}$ concentration showed inhibitory effect on NO production by 38%. Production of IL-$1{\beta}$ and IL-6 were decreased by approximately 56% and 36%, respectively upon GCE treatment at $100{\mu}g/m{\ell}$. Also, production of TNF-${\alpha}$ was decreased by approximately 79% at $100{\mu}g/m{\ell}$. Moreover, the GCE showed inhibitory effects on the expression of the IL-$1{\beta}$, IL-6 and TNF-${\alpha}$ genes in LPS-induced Raw 264.7 cells. Conclusions : From the results above, we conclude that the GCE indicated significant antioxidant effects and induced reduction of pro-inflammatory cytokine.

Vasorelaxant Effect of Prunus yedoensis leaf on Rat Aortic Rings (앵엽(櫻葉) 에탄올 추출물의 혈관이완 효능 및 작용기전에 대한 연구)

  • Lee, Kyungjin;Kim, Kwang-Woo;Heo, Heeseung;Ham, Inhye;Lee, Mi-Hwa;Kim, Bumjung;Bu, Youngmin;Kim, Hocheol;Choi, Ho-Young
    • The Korea Journal of Herbology
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    • v.28 no.4
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    • pp.63-69
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    • 2013
  • Objectives : The purpose of present study was to investigate the vasorelaxant activities and mechanisms of action of the ethanol extract of P. yedoensis leaf (PYL) on isolated rat aortic rings. Methods : Dried P. yedoensis leaves were extracted 3 times with 100% ethanol for 3 h in a reflux apparatus. Isolated rat aortic rings were suspended in organ chambers containing 10 ml Krebs-Henseleit (K-H) solution. The rings were maintained at $37^{\circ}C$ and aerated with a mixture of 95% $O_2$ and 5% $CO_2$. Changes in their tension were recorded via isometric transducers connected to a data acquisition system. Results : PYL relaxed the contraction of aortic rings induced by phenylephrine (PE, 1 ${\mu}M$) or KCl (60 mM) in a concentration dependent manner. However, the vasorelaxant effects of PYL on endothelium-denuded aortic rings were lower than endothelium-intact aortic rings. And the vasorelaxant effects of PYL on endothelium-intact aortic rings were reduced by pre-treatment with $N{\omega}$-Nitro-L-arginine methyl ester (10 ${\mu}M$), methylene blue (10 ${\mu}M$), 1-H-[1,2,4]-oxadiazolo-[4,3-${\alpha}$]-quinoxalin-1-one (10 ${\mu}M$), tetraethylammonium (5 mM). In addition, PYL inhibited the contraction induced by extracellular $Ca^{2+}$ in endothelium-denuded aortic rings pre-contracted by PE or KCl in $Ca^{2+}$-free K-H solution. Conclusions : These results suggest that PYL exerts its vasorelaxant effects via the activation of Nitric Oxide (NO) formation by means of L-arginine and NO-cGMP pathways and via the blockage of receptor operated calcium channels, voltage dependent calcium channels and calcium-activated potassium channels.

The anti-inflammatory effect of Lithospermum Erythrorhizon on lipopolysaccharide - induced inflammatory response in RAW 264.7 cells (LPS로 유도한 RAW 264.7 세포의 염증반응에서 자초(紫草)의 항염증 효과)

  • Choi, Sun-Bok;Bae, Gi-Sang;Jo, Il-Joo;Park, Kyoung-Chel;Seo, Seung-Hee;Kim, Dong-Goo;Shin, Joon-Yeon;Gwak, Tae-Sin;Lee, Jung-Hyun;Lee, Guem-San;Park, Sung-Joo;Song, Ho-Joon
    • The Korea Journal of Herbology
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    • v.28 no.2
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    • pp.67-73
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    • 2013
  • Objective : Lithospermum Erythrorhizon (LE) has been used as an anti-bacterial and anti-inflammatory agent. However, it is unclear that LE aqueous extract could show the anti-inflammatory effects in RAW 264.7cells. The purpose of this study was to investigate the anti-inflammatory effect of aqueous extract from LE on lipopolysaccharide (LPS) - induced inflammatory response. Methods : To measure out the cytotoxicity of LE, we performed the MTT assay. To evaluate the anti-inflammatory effects of LE, we examined the inflammatory mediators such as nitric oxide (NO), prostaglandin E2 ($PGE_2$) and pro-inflammatory cytokines (tumor necrosis factor (TNF)-${\alpha}$, interleukin, (IL)-$1{\beta}$ and (IL)-6) on RAW 264.7 cells. We also examined molecular mechanisms such as mitogen-activated protein kinases (MAPKs) and nuclear factor-B (NF-${\kappa}B$) activation by western blot. Results : Aqueous Extract from LE itself did not have any cytotoxic effect in RAW 264.7 cells. Aqueous extract from LE inhibited LPS-induced productions of inflammatory mediators such as NO, $PGE_2$, and pro-inflammatory cytokines including TNF-${\alpha}$, IL-$1{\beta}$ and IL-6 in RAW 264.7cells. In addition, LE inhibited the phosphorylation of p38 kinases (p38), c-Jun $NH_2$-terminal kinase (JNK), and NF-${\kappa}B$ activation in RAW 264.7 cells. Conclusion : LE down-regulated LPS-induced production of inflammatory mediators through the inhibition of p38, JNK and NF-${\kappa}B$ activation. Taken together, these results could provide the evidence for the anti-inflammatory effects of LE. Therefore, LE may be a novel target in the management of inflammation and help to support a potential strategy for prevention and therapy of inflammatory diseases.

Inhibitory Effect of Water Extract from Dojuksan on LPS-induced Proinflammatory Cytokines Production in RAW 264.7 Cells (LPS로 자극한 RAW 264.7 세포에서 염증성세포활성물질 생산에 미치는 도적산(導赤散) 물 추출의 억제 효과)

  • Kim, Ji-Eun;Kim, Sung-Bae;Kang, Ok-Hwa;Shin, In-Sik;Kang, Suk-Hoon;Lee, Seung-Ho;Kwon, Dong-Yeul
    • The Korea Journal of Herbology
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    • v.28 no.3
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    • pp.53-60
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    • 2013
  • Objectives : DojukSan is known to be effective for treating a urinary diseases and stomatitis. However, there has been a lack of studies regarding the effects of Dojuksan on the inflammatory activities and effector inflammatory disease mechanism about macrophage before is not known. To elucidate the molecular mechanisms of Dojuksan water extract (DJS) on pharmacological and biochemical actions in inflammation, we examined the effect of DJS on pro-inflammatory mediators in lipopolysaccharide (LPS)-stimulated macrophages. Methods : In the present study, pro-inflammatory cytokine production was determined by performing enzyme-linked immunosorbent assay, reverse transcription polymerase chain reaction, and western blot analysis to measure the activation of MAPKs. Cells were treated with 200 ng/mL of LPS 1 h prior to the addition of DJS. Cell viability was measured by MTS assay. The investigation focused on whether DJS inhibited nitric oxide (NO) and prostaglandin E2 ($PGE_2$) productions, as well as the expressions of inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), interleukin-6 (IL-6) and mitogen-activated protein kinases (MAPKs) in LPS-stimulated RAW 264.7 cells. Results : We found that DJS inhibited LPS-induced NO, $PGE_2$ and IL-6 productions as well as the expressions of iNOS and COX-2. Furthermore, DJS suppressed the LPS-induced phosphorylation of p38 MAPK and c-Jun NH2-protein kinase (JNK). Conclusions : These results suggest that DJS has inhibitory effects on LPS-induced $PGE_2$, NO, and IL-6 production, as well as the expressions of iNOS and COX-2 in the murine macrophage. These inhibitory effects occur through blockades on the MAPKs phosphorylation.

The Study of Anti-inflammatory Effect of Hwanggeumjakyak-tang Extract in RAW 264.7 Macrophage (황금작약탕(黃芩芍藥湯)의 RAW 264.7 대식 세포에서의 항염증 효과에 관한 연구)

  • Kim, Ma-Ryong;Kang, Ok-Hwa;Kim, Sung-Bae;Kang, Hee-Jung;Kim, Ji-Eun;Hwang, Hyeong-Chil;Kim, In-Won;Kwon, Dong-Yeul
    • The Korea Journal of Herbology
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    • v.28 no.1
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    • pp.43-50
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    • 2013
  • Objectives : Hwanggeumjakyak-tang (huangqin shaoyao tang, HJT) has been used to treat acute enteritis in traditional oriental medicine. However, there has been a lack of studies regarding the effects of HJT on the inflammatory activities and effector inflammatory disease mechanism about macrophage before is not known. So we examined the effect of HJT water extract on pro-inflammatory mediators in lipopolysaccharide (LPS) - stimulated mouse macrophage, RAW 264.7 cells. Methods : Cells were treated with 2 ug/mL of LPS 1 h prior to the addition of HJT. Cell viability was measured by MTS assay. The production of nitric oxide (NO) was determined by reacting cultured medium with Griess reagent. The expression of cyclooxygenase-2 (COX-2), inducible NO synthase (iNOS) and mitogen-activated protein kinases (MAPKs) was investigated by Western blot, RT-PCR. The content of level of cytokines (prostaglandin (PG) $E_2$, interleukin (IL)-6, IL-12, Tumor necrosis factor-alpha (TNF-${\alpha}$) and monocyte chemoattractant protein-1 (MCP-1)) in media from LPS-stimulated Raw 264.7 cells was analyed by ELISA kit. Results : HJT inhibited the production of NO, $PGE_2$, IL-6 as well as the expressions of iNOS, COX-2 but did not inhibit the production of IL-12, TNF-${\alpha}$, MCP-1 in the murine macrophage, RAW 264.7 cells. HJT also had suppression effects of LPS-induced MAPKs activation Conclusion : These results suggest that HJT has an anti-inflammatory therapeutic potential, which may result from inhibition of MAPK phosphorylation, thereby decreasing the expression of pro-inflammatory genes.

Effect of Polygoni Multiflori Radix Water Extract on the Proinflammatory Mediators in RAW 264.7 Cells Induced by LPS (하수오(何首烏) 물추출물이 LPS로 유발된 RAW 264.7 Cells의 염증인자에 미치는 영향)

  • Jeong, Seong-Yong;Han, Hyo-Sang;Lee, Young-Jong
    • The Korea Journal of Herbology
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    • v.28 no.6
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    • pp.101-109
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    • 2013
  • Objectives : The purpose of this study was to investigate the effects of Polygoni Multiflori Radix Water Extract (PM) on the production of inflammatory mediators in RAW 264.7 mouse macrophages induced by lipopolysaccharide (LPS). Method : We examined effect of PM Extract on the cell viability of RAW 264.7 cells. Futhermore, we investigated anti-inflammatory effect of PM Extract by the production of proinflammatory cytokines such as NO, intracellular calcium, interleukin(IL)-$1{\alpha}$, IL-3, IL-$1{\beta}$, IL-6, interferon inducible protein-10(IP-10), keratinocyte-derived chemokine(KC) and vascular endothelial growth factor(VEGF). Result : No significant changes have been found in the mouse macrophge cell viability by the PM Extract at the concentration of 25, 50, 100 and $200{\mu}g/mL$. The water extract of PM significantly inhibited the production of NO and intracellular calcium in the LPS-induced macrophages at the concentration of 25, 50, 100 and $200{\mu}g/mL$. The water extract of PM significantly inhibited the production of IL-$1{\alpha}$, IL-${\beta}$, IL-3, IP-10, KC, VEGF in the LPS-induced macrophages at the concentration of 50, 100, and $200{\mu}g/mL$; IL-6 at the concentration of 100 and $200{\mu}g/mL$ ; and IL-17 at $200{\mu}g/mL$. Conclusion : The water extract of PM significantly inhibited the production of NO, intracellular calcium, IL-$1{\alpha}$, IL-3, IL-${\beta}$, IP-10, KC, VEGF at the concentration of 50 ㎍/mL or higher in the LPS-induced macrophages with no changes in the cell viability of them. These results suggest that water extract of Polygoni Multiflori Radix has anti-inflammatory effect regulating the production of proinflammatory cytokines in the LPS-induced macrophages.

Inhibition Effects on Oxidative DNA Damage and Anti-inflammatory Effects of Nelumbinis Flos (연꽃의 산화적 DNA 손상 억제 활성 및 항염증 효과)

  • Jeong, Hyung Jin;Park, Yeon Gyeong;Jang, Tae Won;Kim, Do Wan;Jeong, Jin Boo;Park, Jae Ho
    • The Korea Journal of Herbology
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    • v.32 no.3
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    • pp.45-53
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    • 2017
  • Objective : Nelumbo nucifera, its rhizome and semen have been used as a traditional medicine which was studied on antioxidant, hepatoprotective effect, anti-obesity and the others. However, Nelumbinis Flos have not studied. We investigated protective effects on oxidative DNA damage and anti-inflammatory effects of Nelumbinis Flos. Methods : The antioxidant activity was conducted by 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical, 2, 2'-Azino-bis (3-ethylbenzothiazoline-6 sulfonic acid) diammonium salt (ABTS) radical scavenging assay and reducing power assay. Total phenolic content was analyzed. Also, phenolic compounds were detected by HPLC/UV. The inhibitory effect on oxidative DNA damage was determined using ${\Phi}X-174$ RF I plasmid DNA cleavage assay. The anti-inflammatory effect of Nelumbinis Flos was measured by the amount of nitric-oxide (NO) produced and protein levels of iNOS, and COX-2 in LPS induced RAW 264.7 cells. Results : The results of DPPH and ABTS radical scavenging activity at $200{\mu}g/m{\ell}$ of extraction were $97.02{\pm}0.88%$ and $96.42{\pm}0.25%$. Reducing power (fold of L-ascorbic acid as control) was $100.14{\pm}0.31$ at $200{\mu}g/m{\ell}$. Total phenol content was $8.70{\pm}0.02mg/g$. Chlorogenic acid, catechin and epicatechin were found by HPLC. Nelumbinis Flos has inhibitory effect in dose-manner against oxidative DNA damage. In addition, it showed the anti-inflammatory effect by suppression of NO production as well as protein levels of iNOS, and COX-2. Conclusion : This study suggested that Nelumbinis Flos showed potential antioxidant and suppression activities of various factors were related in NO produced. Therefore, Nelumbinis Flos as natural plant resources that may help reduce inflammation and alleviate DNA damage.

Protective Effect of Dioscoreae Rhizoma Extracts in MIA-induced Rat. (MIA로 유발된 골관절염 동물모델에서 산약 추출물의 골관절염 개선 효과)

  • Kim, Min Ju;Park, Hae-Jin;Kim, Kyeong Jo;Lee, Jin A;Shin, Mi-Rae;Roh, Seong-Soo
    • The Korea Journal of Herbology
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    • v.34 no.4
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    • pp.27-35
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    • 2019
  • Objectives : Osteoarthritis is characterized by degeneration of articular cartilage, which is characterized by chronic pain, stiffness and decrease range of motion. The present study was designed to compare the therapeutic effect of Dioscoreae Rhizoma water extract (DRW) and Dioscoreae Rhizoma 30% ethanol extract (DRE) on the monosodium iodoacetate (MIA)-induced osteoarthritis rats. Methods : Osteoarthritis was induced by injection of MIA ($50{\mu}{\ell}$ with $80mg/m{\ell}$) into the knee joint cavity of rats. After adaptation period for seven days, rats were divided by 5 groups (n=10/group): normal group, control group, positive control (indomethacin 5 mg/kg), DRW 200 mg/kg treated group, DRE 200 mg/kg treated group (n=10/group). The hind paw weight distribution was measured with the changes of reactive oxygen species (ROS), peroxynitrite ($ONOO^-$) in articulation tissue. Also, the cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), tumor necrosis factoralpha ($TNF{\alpha}$), interleukin-6 (IL-6), matrix metalloproteinase-1 (MMP-1), and tissue inhibitor of metalloproteinases-1 (TIMP-1) were investigated by western blot analysis. Results : The administration of DRW and DRE significantly decreased the hind paw weight distribution. The ROS and $ONOO^-$ levels of cartilaginous tissue were significantly decreased in DRW and DRE compared to control group. The results showed that DRE decreased inflammatory cytokines such as iNOS and $TNF{\alpha}$. Also DRE decreased MMP-1 and increased TIMP-1. Conclusions : Based on the above results, Dioscoreae Rhizoma extract seems to have the therapeutic effect on osteoarthritis via suppression of inflammation.