• 제목/요약/키워드: MTT method.

검색결과 420건 처리시간 0.031초

Effects of Salviae Miltiorrhizae Radix Hot Aqueous Extract on Nitric Oxide and Prostaglandin E2 Production and on 1,1-diphenyl-2-picryl hydrazyl Radical Scavenging in Macrophages

  • Yeo, In Ho;Lee, Cham Kyul;Lee, Eun Yong
    • 대한약침학회지
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    • 제17권1호
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    • pp.7-12
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    • 2014
  • Objectives: The objective of this study is to investigate the effects of Salviae Miltiorrhizae Radix hot aqueous extract on nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) production and on 1,1-diphenyl-2-picryl hydrazyl (DPPH) free-radical scavenging in macrophages. Methods: Salviae Miltiorrhizae Radix (300 g) was heated at $100^{\circ}C$ with distilled water (2 L) for 4 hours. The extract was filtered and concentrated to 100 mL by using a rotary evaporator, was frozen at $-80^{\circ}C$, and was then freeze-dried by using a freezing-drying system. The RAW 264.7 macrophage was subcultured by using $10-{\mu}g/mL$ lipopolysaccharide (LPS). In order to evaluate cytotoxicity, we performed 3-(4,5-dimrthylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays and measured the cell viability. The NO production was measured by using Griess assays, and the $PGE_2$ production was measured by using enzyme immunoassays. The antioxidant activity, the 1,1-diphenyl-2-picryl hydrazyl (DPPH) free-radical scavenging capability, was measured by using the DPPH method. Results: Cell viability with the 1-, 5-, 25-, 125- and $625-{\mu}g/mL$ Salviae Miltiorrhizae Radix hot aqueous extract was not significantly decreased compared to the cell viability without the extract. When 125 and $625{\mu}g/mL$ of Salviae Miltiorrhizae Radix hot aqueous extract were used, nitric oxide (NO) production in LPS-stimulated RAW 264.7 macrophages was significantly inhibited compared to that in the control group. When 25, 125, and $625{\mu}g/mL$ of Salviae Miltiorrhizae Radix hot aqueous extract were used, $PGE_2$ production in LPS-stimulated RAW 264.7 macrophages was significantly inhibited compared to that in the control group. The 125- and $625-{\mu}g/mL$ Salviae Miltiorrhizae Radix hot aqueous extracts had high DPPH free-radical scavenging capabilities in RAW 264.7 macrophages. Conclusion: This study indicates that Salviae Miltiorrhizae Radix hot aqueous extract suppresses NO and $PGE_2$ production and improves DPPH free-radical scavenging capability. Thus, it seems that Salviae Miltiorrhizae Radix hot aqueous extract may have an anti-inflammation effect and antioxidant activity.

사람 Mast cell에서의 케모카인에 대한 백연치효탕(白蓮治哮湯)의 효과(效果) (Effect of Baekryunchihyo-tang(白蓮治哮湯) on Expression of Chemokines in Human Mast Cell Line (HMC-I))

  • 정희재;이형구;정승기;이형철
    • 대한한방내과학회지
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    • 제25권4호
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    • pp.289-299
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    • 2004
  • Background : Although the pathophysiology of asthma has been reported, its mechanism has not been fully elucidated. The mast cell is an effector cells in allergic inflammation and secretes a number of chemokines. Chemokines are important for the recruitment of leukocytes to sites of infection, which is essential in host defense. Chemokines also contribute to the pathogenesis of several disorders such as asthma, chronic bronchitis, atopic dermatitis, allergic rhinitis, and rheumatoid arthritis. Objective : In this study, the aim was to identify the effect of Baekryunchihyo-tang(白蓮治哮湯) on expression of chemokines. This was examined by RT-PCR using the human mast cell line (HMC-l) Materials and Methods : HMC-l cells were used, which is known to secrete and express chemokines. In order to investigate the protective effect of Baekryunchihyo-tang(白蓮治哮湯), HMC-l cells were incubated with pretreatment of Baekryunchihyo-tang(白蓮治哮湯) for 24 hrs. RT-PCR analyses of chemokine genes of cells pretreated with Baekryunchihyo-tang(白蓮治哮湯) showed that expressions of IL-8, $MIP-l{\beta}$, and RANTES genes in these cells were lower and $MIP-l{\alpha}$ showed a similar pattern compared to the calcium ionophore-treated group. In addition, cell cytotoxicity concentration measurements were performed by MTT assay method. Results : After stimulation with 1 uM calcium ionophore A23178 for 2 hrs, IL-8, major one of CXC chemokines, was highly expressed, and expression of $MIP-l{\beta}$ and RANTES (CC chemokines) increased, while expression of $MIP-l{\alpha}$ did not change. The cell cytotoxicity of Baekryunchihyo-tang(白蓮治哮湯) with treatments at various concentrations and times was not observed, respectively. Conclusion : This study suggests that Baekryunchihyo-tang(白蓮治哮湯) has dose-dependent effects on mRNA expression of IL-8(CXC chemokines), $MIP-l{\beta}$ and RANTES(CC chemokines) in human mast cellline(HMC-l). So these herbal medicines may inhibit the inflammatory process of asthma. Advanced studies are required to investigate the mechanism of inhibition by herbal medicine in the asthma model. This study provides basic data on the possibility of the clinical treatment of Baekryunchihyo-tang(白蓮治哮湯) for allergic disorders.

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련화향(蓮花香) 정유액이 glioma cell에 미치는 효과 (Effects of the Essential Oil of Nelumbo nucifera Flower on Glioma Cells)

  • 김인자;이주연;최방섭;김근우;구병수
    • 동의신경정신과학회지
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    • 제19권2호
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    • pp.111-122
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    • 2008
  • Objective : Herb medicines are potential sources of useful edible and medicinal plants. They are used as a drug because of their various biological activities such as immunomodulatory, antiviral, and antitumor functions. Nelumbo nucifera have been applied in Chinese herbal prescriptions to improve tissue inflammation. However, it has not been elucidated on the effect of the flower of Nelumbo nucifera in cells. Method : In the present study, to examine the effect of that on glioma cells, U87, the essential oil was extracted from the flower of Nelumbo nucifera (NN essential oil). U87 cells were exposed to different concentrations of 2-40 ug/ml of NN essential oil in ethanol. Cell viability was measured by MTT assay at 24 h. To find out the intracellular target signal molecule(s) for this antiproliferative activity of NN essential oil, phosphorylation of Akt, ERM, MAPK or p38 proteins were examined by Western blot analysis. To study long term effect of NN essential oil in U87 cells, the image of cells treated with NN essential oil for 4 days were obtained. Results and Conclusion : NN essential oil was shown to exhibit antitumor activity in glioma cells, at a broad range of concentrations of 10-40 ug/ml. The phosphorylation of Akt and Endoplasmic Reticulum Matrix (ERM) proteins which known to be involved in the cell death, were gradually decreased to 2 hours after addition 20 ug/ml of NN essential oil. However, the phosphorylation of mitogen-activated protein (MAPK) and p38 was found to increase in NN essential oil treated cells. NN essential oil treated cells showed decreased glioma cell number. These results provide a possible NN essential oil-induced inhibitory signal for tumor cell proliferation that is initiated by the decrease in Akt activity. Moreover, it is likely that the activation of p38 is required for the NN essential oil-induced inhibition of tumor proliferation.

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티트리 에센셜오일의 생물활성 및 주요 성분 분석 (Bioactivity and Chemical Composition of the Essential oil of Tea Tree (Melaleuca alternifolia))

  • 양선아;전상경;이은정;임남경;정지영;이인선
    • 생명과학회지
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    • 제18권12호
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    • pp.1644-1650
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    • 2008
  • 티트리 에센셜 오일은 호주 원주민들의 전통적인 피부 소독제나 치료제로 널리 사용되어 왔으며, 항균효과와 주요성분 등 많은 보고가 있으나 추출 방법이나 사용 부위 등에 따라 효능의 차이를 보인다. 본 연구에서는 아로마테라피 등에 현재 많이 이용되고 있는 시판 티트리 오일의 성분과 효능을 평가하여, 다른 에센셜 오일과의 비교 이용을 용이하게 하고자 하였다. 티트리 오일의 주요성분은 GC-MS 분석에 의하여 ${\beta}$-terpinene (20.87%), ${\alpha}$-pinene (17.60%), p-cymene (11.23%), 3-carene (10.40%), trans-anethole (8.47%), limonene (4.65%)으로 밝혀졌으며, 5% 이하의 농도에서 3시간 미만까지는 피부세포에 독성이 없었다. 오일의 라디컬 소거능을 알아본 결과, DPPH와 ABTS의 양라디컬에 대하여 강한 소거능을 나타내어 강한 항산화능을 시사했다. 또한, 오일의 direct contact와 vapor-phase의 항균활성을 disc diffusion법으로 스크리닝 한 결과, direct contact 활성의 경우 그람음성균에 대하여 높은 활성을 나타내었으며, vapor는 S. aureus에 대하여 강한 효과를 나타내었다. 본 연구에서 실제 많이 사용되는 티트리 오일의 성분과 생물활성을 측정함으로써 허브 오일들의 정확한 선택과 활용을 위한 기본적인 결과를 얻었다.

군소내장 분획물의 in vitro에서의 암세포 성장억제 및 quinone reductase 유도 활성 증가 효과 (Growth Inhibitory and Quinone Reductase Activity Stimulating Effects of Internal Organs of Aplysia kurodai Fractions on Cancer Cell Lines In vitro)

  • 신미옥
    • 생명과학회지
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    • 제20권6호
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    • pp.877-884
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    • 2010
  • 본 연구에서는 군소내장을 각 용매별로 분획하여 암세포 성장 억제 효과와 quinone reductase (QR) 유도 활성증가 효과를 알아보았다. 간암 세포인 HepG2, 유방암 세포주인 MCF-7, 대장암 세포주인 HT-29 그리고 피부암 세포주인 B16-F10 를 이용하여 암세포 성장 억제 효과를 실험한 결과 모든 세포주에서 AKMM층에서 농도 의존적으로 가장 높은 암세포 성장 억제 효과를 나타내었다. 그리고 다음으로 AKMB층, AKMH층 및 AKMA층의 순이었다. 그리고 4종의 암세포주 중에서 B16-F10 세포주가 가장 높은 암세포 성장 억제 효과를 나타내어 특히 피부암에 대한 예방효과가 기대되어진다. 또한 암 예방 효과를 알아보기 위하여 4종의 암세포 중 유일하게 quinone reductase를 가지고 있는 인체 간암세포주인 HepG2를 이용하여 QR 유도 활성 증가 효과를 측정한 결과, 암세포 성장 억제 효과에 있어서 가장 높은 효과를 나타낸 AKMM층에서 가장 높은 QR 유도 활성 증가 효과를 나타내었다. 그러나 다른 분획층에서는 QR 유도 활성 증가 효과를 거의 볼 수 없었다. 이상으로 암세포 성장 억제 효과와 QR 유도 활성 효과에서 모두 methanol 분획층인 AKMM층에서 가장 높게 나타났으므로 이 분획층에 유효한 생리활성 물질이 함유되어 있을 가능성이 추정되어진다. 따라서 폐기되어지는 군소부산물인 내장을 이용한 암 예방 관련 기능성 식품의 개발 가능성이 기대되어지며, 이를 위한 AKMM분획층에 대한 더욱더 심도 높은 집중적인 연구가 요구되어진다.

대조 추출물분획이 치은 섬유아세포의 생물학적 활성화에 미치는 영향 (EFFECT OF ZIZYPHI FRUCTUS EXTRACT ON THE BIOLOGICAL ACTIVITY OF GINGIVAL FIBROBLAST)

  • 양창호;이용무;조기영;배기환;정종평
    • Journal of Periodontal and Implant Science
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    • 제24권1호
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    • pp.144-154
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    • 1994
  • Final goal of periodontal treatment is to reconstruct the destructed periodontal tissue as well as to remove the necrotic pathologic elements. The purpose of this study is to investigate on the effect of Zizyphi extract to the inhibitory ability on collagenolytic activity of P gingivalis, biologic activity of gingival fibroblasts, and on the collagen and protein synthesis of gingival fibroblasts. Gingival fibroblast from giniva of first bicuspids from patient for orthodontic treatment were used and cultured. For the measurement of inhibitory ability of collagenolytic activity, crude enzyme was extracted and used on the basis of modified Ono's method. On the inhibition of collagenolytic enzyme from herbal extracts, collagenokit CLN-100 were used. The cellular activity of gingival fibroblast, were studied using MTT solution and measured optical density on 570mm by ELISA reader. To measure the effects on the ability of whole protein and collagen synthesis, cell membrane was destructed with ultrasonic grinder after culturing, centrifuged and counted by liquid scintilation counter. The inhibitory effects on producing of $IL-l{\beta}$ by monocyte, after promotion of producing $IL-l{\beta}$ by LPS, were compared with the mixture of herbal extracts and other drugs using thymocyte stimulation assay. About inhibitory effects of $PGF_2$. by gingival fibroblasts, herbal extract was compared with the addition of the other control groups using enzyme imunoassay. On the inhibition of collagenolytic activity by P. gingivalis, benzene extracts showed the most efficient inhibitory effects among the $19{\mu}g/ml$ of the compared extracts and 40.5% by Tetracycline. On the cellular activity promoting effects, compared extracts showed a bit of more effects than PDGF of $100{\mu}g/ml$ concentration and IGF of $20{\mu}g/ml$ concentration. All of the PDGF, IGF, Zizyphi Fructus extract should increase in collagen synthesis, but especially 70% ethylalcohol extracts of Zizyphi Fructus showed comparably high effects among the compared extracts. Effects on whole protein synthesis were slightly increased on every extract but especially 70% ethylalcohol extract showed significantly effective than any other estract. On the inhibitory effects of Zizyphi Fructus $IL-l{\beta}$ production by monocyte, compared extracts showed 70% of highly inhibitory effect than that of 60% inhibition effects on controlled group and each extracts showed no significant difference. In $PGF_2$ production inhibitroy effect of Zizyphi Fructus gingival fibroblasts, Herbal extracts showed 70% of inhibition comparing with tat of 90.2% of controlled group, but each extracts showed similar effects excluding the $H_2O$ extracts. These results suggested that Zizyphi Fructus might be useful medicine for inhibition of inflammatory mediator including $IL-l{\beta}$ and $PGF_2$.

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후박 및 대조추출혼합물이 골조직 재생에 미치는 영향 (Tissue regenerative activity of Magnolia and Zizyphi fructus extract mixtures)

  • 이용무;구영;배기환;정종평
    • Journal of Periodontal and Implant Science
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    • 제27권1호
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    • pp.165-177
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    • 1997
  • The purpose of this study was to perform on the biological activity of Magnolia and Zizyphi fructus extract mixtures on the wound healing of defected rat calvaria. For the determination of the mixture ratio of two extracts for oral administration, preliminary experiments were performed with the mixture combination of 2000 and $3000{\mu}g/ml$ of Magnolia extract, and also 20, 30, 200, 300, 2000 and $3000{\mu}g/ml$ of Zizyphi fructus extract, respectively and divided into 6 groups. The combination of extracts mixture were tested on the enhancing effect of cellular activity. The effect of the extracts mixture on the cellular activity was evaluated using MTT method and measured on the results with optical density by ELISA reader. The ability to tissue regeneration of the extracts mixture was performed by measuring new bone and new connective tissue regeneration on the 5mm defected rat calvaria for 1, 2 and 3 weeks after oral administration of 2 different dosages groups : 10:1(0.1g/kg) and 10:1(0.5g/kg). It was employed the same dosages of unsaponifiable fraction of Zea Mays L as positive controls. Each group of rat was sacrificed and en bloc section for histological examination. The effect on the cellular activity of each mixture ratio showed significantly higher in $2000{\mu}g/ml$ of Magnolia extract and $200{\mu}g/ml$ of Zizyphi fructus extract group to compare with other groups. These preliminary results showed that appropriate mixture ratio of two extracts was 10:1 of Magnolia and Zizyphi fructus extract. Histological examination on the activity of tissue regeneration of each group showed that 2weeks and 3weeks specimens of 0.5g/kg of 10:1 extract mixture of Magnolia and Ziziphi fructus administrated rat calvaria revealed significantly more osteoid and new bone formation of defected calvaria with unification of defected area than the specimens of any other negative and positive controls. Even though the specimen administrated the same dosages of unsaponifiable fraction of Zea Mays L, positive controls, showed the trend that they promote significantly the repair of calvarial defect, their bone reparative activities were less inductive than the same dosages of Magnolia and Ziziphi fructus extract mixture. These results implicated that the mixture of Magnolia and Zizyphi fructus extracts should be highly effective on the wound healing of bony defected site and might have potential possibilities as an useful drug to promote periodontal tissue regeneration.

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시판 중인 구강청결티슈의 세포 독성 관찰 (The Cytotoxic Effect of Oral Wet Wipes on Gingival Cells)

  • 정임희;박지현;이민경;황영선
    • 치위생과학회지
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    • 제18권2호
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    • pp.76-84
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    • 2018
  • 구강위생을 위한 간편성과 편리성 때문에 영유아에서 건강취약자 및 일반인에 이르기까지 구강청결티슈 사용이 증가되고 있다. 또한 기본 성분인 정제수 외에도 구강위생에 도움이 될 목적으로 다양한 기능 성분이 첨가된 구강청결티슈들이 시판되고 있다. 하지만 함유 성분에 대한 제공정보가 부족하고 함유량 기준이 마련되지 못하고 있어 구강 환경이 예민한 영유아 등에게 적용하기 위해서는 이들 제품에 대한 연구자료가 필요할 것으로 보인다. 본 연구에서는 구매도가 높은 시판 구강청결티슈 5종에 대한 구강세포 안전성을 확인함으로써 제품사용 시 유의해야 할 정보를 제공하고자 하였다. 시험 결과 피셔프라이스와 닥터케네디 제품 성분은 구강미생물 S. mutans 및 A. actinomycetemcomitans에 대한 항균작용을 나타냈지만 구강상피세포 및 구강섬유세포에도 독성을 나타냈다. 항균작용이 제한적인 궁중비책, 마이비, 아이수 제품 성분은 구강상피세포 및 구강섬유세포에 대한 독성도 낮았다. 피셔프라이스와 닥터케네디 제품 성분에 의한 구강세포 독성은 G2/M phase에서 세포주기 진행 억제와 세포자멸 유도에 의한 것으로 분석되었다. 따라서 구강 환경이 예민한 영유아 등을 대상으로 한 반복적이고 빈번한 사용은 구강세포 독성의 가능성을 높일 수 있다.

천연 암반 심해수의 항산화 활성 및 면역반응에 대한 연구 (Study of Deep Ground Sea-Like Water on Antioxidant Activity and the Immune Response in RAW264.7 Macrophages.)

  • 김유정;정일선;송효주;최은영;최인순;최영주
    • 생명과학회지
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    • 제18권3호
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    • pp.329-335
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    • 2008
  • 해양성 심층수(KDSW)는 해양심층수(DSW)와 유사한 mineral 조성을 가지고 있어 식품 및 의약분야에서 그 유용성에 대한 관심이 증대되고 있다. 본 연구에서는 KDSW 및 탈염한 Danasoo의 항산화력, 면역활성, 함암 및 당뇨에 대한 효과를 연구하였다. KDSW와 Danasoo의 항산화 활성은 DPPH radical scavenging 활성, SOD-like 활성 및 PCL 법에 의하여 항산화 활성을 조사하였다. KDSW와 Danasoo의 항산화 활성은 첨가된 농도에 의존적으로 증가하였으며, 특히 PCL법에 의한 KDSW와 Danasoo의 항산화능은 85.32과 14.02(nmol of ascorbic acid equivalent/ml)로 높은 항산화 활성을 나타내었다. Macrophage RAW 264.7 cell에서 LPS에 의하여 유도된 NO 활성은 Danasoo (20%)를 첨가에 의하여 약 30%정도의 NO 합성이 저해되었으며, 이 농도에서 세포 독성이 없는 것으로 MTT assay에 의하여 확인하였다. NO는 nitric oxide synthase에 의하여 합성되며, tumor성장 및 angiogenesis에서 중요한 역할을 하는 것으로 밝혀져 있으며, Danasoon (20%)는 위암세포와 폐암세포의 iNOS 발현을 현저히 감소시켰다. STZ에 의하여 유도된 당뇨 쥐의 혈당량은 Danasoo (20%) 식이에 의하여 대조구와 유사한 당뇨효과를 나타내었다. 이러한 결과들은 KDSW가 우수한 biological 활성을 가지고 있으며, 또한 천연 기능성 제품생산을 위한 소재로 사용될 수 있는 높은 가능성올 시사하고 있다.

Anti-oxidant and Anti-inflammatory Effects of Acanthopanacia Cortex Hot Aqueous Extract on Lipopolysaccharide(LPS) Simulated Macrophages

  • Jo, Na Young;Roh, Jeong Du
    • Journal of Acupuncture Research
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    • 제31권1호
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    • pp.131-137
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    • 2014
  • Objectives : This study is to investigate the effects of Acanthopanacis Cortex hot aqueous extract on nitric oxide(NO), prostaglandin E2(PGE2) production and DPPH(1,1-diphenyl-2-picryl hydrazyl) radical scavenging activity in macrophages. Methods : Acanthopanacis Cortex(200 g) was heated at $100^{\circ}C$ with distilled water(2 L) for 4hrs. The extract was filtered and concentrated to 100 ml using a rotary evaporator and was frozen at $-80^{\circ}C$, then was freeze-dried. The RAW 264.7 macrophages were subcultured. In order to evaluate cytotoxicity, MTT assay was performed. Experimental groups were divided into five(control, AC 25, 50, 100 and 200 ${\mu}g/ml$) and we measured cytotoxicity. The concentrations of NO were preprocessed by Griess assay. The RAW 264.7 macrophages was pretreated by 10 ${\mu}g/ml$ LPS and experimental groups were divided into five and we measured NO production. The concentrations of $PGE_2$ were measured by enzyme immunoassay. The RAW 264.7 macrophages was pretreated by 10 ${\mu}g/ml$ LPS. Experimental groups were divided into five and we measured $PGE_2$ production. Antioxidant activity was measured by the DPPH method. experimental groups were divided into four(AC 25, 50, 100 and 200 ${\mu}g/ml$) and we measured DPPH radical scavenging activity. Results : 1. Viability of RAW 264.7 macrophages did not significantly decrease in 25, 50 and 100 ${\mu}g/ml$ Acanthopanacis Cortex hot aqueous extract compared to control group. 2. NO production in LPS-stimulated RAW 264.7 macrophages significantly inhibited in 100, 200 ${\mu}g/ml$ Acanthopanacis Cortex hot aqueous extract compared to control group. 3. $PGE_2$ production in LPS-stimulated RAW 264.7 macrophages significantly inhibited in 100, 200 ${\mu}g/ml$ Acanthopanacis Cortex hot aqueous extract compared to control group. 4. DPPH radical scavenging capability of Acanthopanacis Cortex hot aqueous extract in RAW 264.7 macrophages had the high level in 100, 200 ${\mu}g/ml$. Conclusion : According to the results, Acanthopanacis Cortexx hot aqueous extract has ability to suppress NO, $PGE_2$ production and improve DPPH free radical scavenging activity. So Acanthopanacis Cortex hot aqueous extract may have an anti-inflammation effect and antioxidant activity.