• Title/Summary/Keyword: RAW 264.7 Cell Line

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Purification and Identification of Cytotoxic Compounds from the Root of Rumex crispus L. (소리쟁이 뿌리로부터 세포독성물질 분리 및 정제)

  • Cho, Yong Beom;Kim, Jae Yeon;Kwon, Nam Woo;Hwang, Bang Yeon;Kim, Jun Gu;Woo, Sun Hee;Lee, Moon Soon
    • Korean Journal of Medicinal Crop Science
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    • v.27 no.3
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    • pp.208-217
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    • 2019
  • Background: In the present study, we identified two cytotoxic compounds from the root of Rumex crispus L. using a bioassay-based method. Methods and Results: Compared with the other fractions, the diethyl ether ($Et_2O$) fraction of R. crispus root extract exhibited the strongest of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging effect [scavenging concentration 50% $(SC_{50})=63.8{\pm}1.47{\mu}g/m{\ell}$], nitric oxide (NO) production inhibitory effect on the mouse macrophage cell line RAW264.7 [inhibitory concentration 50% $(IC_{50})=60.9{\pm}7.52{\mu}g/m{\ell}$] and cytotoxicity effect on the human hepatoma cell line, HepG2 [lethal concentration 50% $(LC_{50})=115.4{\pm}1.86{\mu}g/m{\ell}$]. According to the bioassay-based method, two cytotoxic compounds were purified from the $Et_2O$ fraction by using column chromatography and preparative high performance liquid chromatography (prep-HPLC). These two compounds were identified as parietin and chrysophanol by using nuclear magnetic resonance (NMR) and liquid chromatography quadruple time of flight mass spectrometry (LC-QTOF-MS). In addition, both parietin and chrysophanol exhibited a cytotoxicity effect on HepG2 cells, their $LC_{50}$ values were $169.1{\pm}17.67{\mu}M$ and $111.5{\pm}6.62{\mu}M$, respectively. Conclusions: Parietin and chrysophanol isolated from the $Et_2O$ fraction of the R. crispus root extract showed cytotoxicity in HepG2 cell.

The Anti-inflammatory Effect of Angelica gigas Nakai Supercritical Carbon Dioxide Selective Extracts (참당귀 초임계 이산화탄소 선택 추출물의 항염 효능)

  • Park, So Hyeon;Lee, Kwang Won;Park, Shinsung;Shin, Moon Sam;Park, Su In
    • The Journal of the Convergence on Culture Technology
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    • v.8 no.6
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    • pp.849-854
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    • 2022
  • In this study, the anti-inflammatory effect of supercritical carbon dioxide selective extracts, which extract decursin and decursinol angelate, the vital active ingredients of Angelica gigas Nakai, in high yield, was measured compared to that of ethanol extracts. To measure the anti-inflammatory effect, the production of nitric oxide(NO), an inflammatory mediator, and interleukin(IL)-6 and IL-8, inflammatory cytokines, was measured. NO production was measured by Griess assay on Raw 264.7 cells induced inflammatory response by lipopolysaccharide(LPS), and IL-6 and IL-8 production was measured by enzyme-linked immunoadsorbent assay(ELISA) on human keratinocyte cell line(HaCaT) cells induced inflammatory response by tumor necrosis factor(TNF)-α. The amount of NO production was suppressed outstandingly by the supercritical carbon dioxide extracts compared to the ethanol extracts. The amount of IL-6 and IL-8 production was increased by the ethanol extracts, whereas statistically significantly inhibited by supercritical carbon dioxide extracts at the concentration of 6.25 ㎍/mL(P<0.01). Through these results, we confirmed that the supercritical carbon dioxide selective extracts of Angelica gigas Nakai could be used as an anti-inflammatory cosmeceutical material to alleviate atopic dermatitis.

Ethanol Extracts from Astilbe chinensis (Maxim.) Franch. Et Savat. Exhibit Inhibitory Activities on Oxidative Stress Generation and Viability of Human Colorectal Cancer Cells (노루오줌 에탄올 추출물의 산화스트레스 및 대장암 세포 억제활성)

  • Nho, Jong Hyun;Jang, Ji Hun;Jung, Ho Kyung;Lee, Mu Jin;Sim, Mi Ok;Jeong, Da Eun;Cho, Hyun Woo
    • Korean Journal of Medicinal Crop Science
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    • v.26 no.2
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    • pp.141-147
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    • 2018
  • Background: Astilbe chinensis (Maxim.) Franch. Et Savat. is a plant belonging to Saxifragaceae family and contains various active ingredients including astilbin and bergenin. It has been used as a traditional Korean medicine to improve fever, pain, and cough. Recently, a number of Korean medical resources have been studied for cancer and inflammation treatment, but A. chinensis (Maxim.) Franch. Et Savat. has not yet been investigated. Consequently, this study investigated the inhibitory effect of ethanol extracts from A. chinensis (Maxim.) Franch. Et Savat. (ARE) on oxidative stress and colorectal cancer using RAW264.7 and the human colorectal cancer cell line HCT-116. Methods and Results: In total, $500{\mu}g/m{\ell}$ ARE reduced cell viability by $38.96{\pm}1.32%$, and increased caspase-3 activity by $133.08{\pm}3.41%$ in HCT-116 cells. Moreover, TUNEL signaling and the early apoptosis ratio ($34.56{\pm}1.67%$) increased by $500{\mu}g/m{\ell}$ ARE treatment. $H_2O_2$-induced oxidative stress and cell death were diminished by $500{\mu}g/m{\ell}$ ARE treatment through decreasing ROS (reactive oxygen species). Conclusions: The inhibitory effects of ARE against human colorectal cancer cells is mediated by apoptosis and caspase-3 activation, and $H_2O_2$-induced ROS generation and cell death are decreased by ARE treatment in RAW264.7 cells. However, further study is required to explore how ARE treatment is involved in the signaling pathway to decrease ROS.

Inhibitory Effect of an Ethanol Extract of Inulae Flos on Nitric Oxide Production, Oxidative Stress and Human Colorectal Cancer Cell Lines (선복화 에탄올 추출물의 Nitric Oxide 생성, 산화스트레스 및 대장암 세포 억제효과)

  • Nho, Jong Hyun;Jung, Da Eun;Jung, Ho Kyung;Lee, Mu Jin;Jang, Ji Hun;Sim, Mi Ok;Jung, Ja Kyun;Cho, Hyun Woo
    • Korean Journal of Medicinal Crop Science
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    • v.26 no.1
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    • pp.19-25
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    • 2018
  • Background: Inula japonica Thunb. is a plant belonging to the family compositae. Inulae flos (flower of I. britannica var. chinensis Regal.) is the dried flower of I. japonica Thunb. and contains various flavonoids (patulitrin, nepitrin and kaempferol), which have been utilized in traditional oriental medicine to treat nausea, phlegm, and coughs. However, ethanol extract of I. britannica (IJE) has not been previously studied for its use in cancer treatment, and its effects on oxidative stress, or inflammation. Thus, the present study investigated the anti-oxidant, anti-inflammatory, and anti-colorectal cancer effects of IJE using RAW264.7 and HCT-116 cells, which are human colorectal cancer cell line. Methods and Results: IJE contained flavonoids ($80.95{\pm}5.3mg/g$) and polyphenols ($310.53{\pm}10.6mg/g$). Moreover, it reduced lipopolysaccharide (LPS)-induced nitric oxide (NO) production and $H_2O_2$-induced oxidative stress by decreasing reactive oxygen species (ROS) levels. Additionally, the $500{\mu}g/m{\ell}$ IJE treatment increased caspase-3 activity and apoptotic cell death in HCT-116 cells. Conclusions: These results demonstrate that the anti-cancer effect of IJE against human colorectal cancer cells involves caspase-3 activation and apoptotic cell death. IJE also inhibited LPS-induced NO production, and $H_2O_2$-induced oxidative stress in RAW264.7 cells. However, further studies are required to explore how IJE treatment regulates signal transduction in NO and ROS production.

Anti-colorectal Cancer and Anti-oxidant Activities of Rubiae radix Ethanol Extract in vitro (천초근 에탄올 추출물의 항산화 효능 및 대장암 세포 억제 효과)

  • Nho, Jong Hyun;Sim, Mi Ok;Jung, Ho Kyung;Lee, Mu Jin;Jang, Ji Hun;Jung, Da Eun;Sung, Tae Kyoung;An, Byeong Kwan;Cho, Hyun Woo
    • Korean Journal of Plant Resources
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    • v.31 no.2
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    • pp.102-108
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    • 2018
  • Rubiae radix is root of Runia akane Nakai, it has been used to hemostasis and blood stasis in Korean and China. This study investigated that anti-oxidant and anti-colorectal cancer effect of ERA (ethanol extract of Rubiae radix) and WRA (water extract of Rubiae radix) using RAW 264.7 (murine macrophage from blood) and HCT-116 cells (human colorectal cancer cell line). ERA contained polyphenol ($45.77{\pm}2.03mg/g$) and flavonoid ($22.82{\pm}1.33mg/g$). $500{\mu}M$ $H_2O_2$-induced ROS generation was diminished by $500{\mu}g/m{\ell}$ ERA treatment in RAW 264.7 cells, but not WRA (125, 250, and $500{\mu}g/m{\ell}$). Moreover, caspase-3 activity and DNA fragmentation increased by $500{\mu}g/m{\ell}$ ERA treatment during apoptotic cell death in HCT-116. Results demonstrated that anti-cancer effect of ERA against human colorectal cancer cells is mediated apoptotic cell death and DNA fragmentation through caspase-3 activation. However, further study is required to what active ingredient of ERA are important for anti-oxidant and anti-colorectal cancer effect in vivo.

The Beneficial Effects of Extract of Pinus densiflora Needles on Skin Health (솔잎추출물의 피부건강 개선효과)

  • Choi, Jieun;Kim, Woong;Park, Jaeyoung;Cheong, Hyeonsook
    • Microbiology and Biotechnology Letters
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    • v.44 no.2
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    • pp.208-217
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    • 2016
  • Pinus densiflora Sieb. et Zucc. (P. densiflora) contains several phenolic compounds that exhibit biological activities, such as antimicrobial, antioxidant, and antihypertensive effects. However, the anti-inflammatory effect of P. densiflora on skin has rarely been reported. Malassezia furfur (M. furfur) is a commensal microbe that induces skin inflammation and is associated with several chronic disorders, such as dandruff, seborrheic dermatitis, papillomatosis, and sepsis. The aim of our study was to identify the anti-inflammatory effects of P. densiflora needle extracts on skin health subjected to M. furfur-induced inflammation. The methanolic extract of the pine needles was partitioned into n-hexane, EtOAc, n-BuOH, and water layers. We measured the anti-inflammatory effects (in macrophages) as well as the antioxidant, antifungal, and tyrosinase inhibitory activity of each of these layers. The antioxidant activity of the individual layers was in the order EtOAc layer > n-BuOH layer > water layer. Only the n-BuOH, EtOAc, and n-hexane layers showed antifungal activity. Additionally, all the layers possessed tyrosinase inhibition activity similar to that of ascorbic acid, which is used as a commercial control. The EtOAc layer was not cytotoxic toward the RAW 264.7 cell line. Interleukin 1 beta and tumor necrosis factor (TNF)-α expression levels in M. furfur-stimulated RAW 264.7 cells treated with the EtOAc layer were decreased markedly compared to those in cells treated with the other layers. Taken together, we believe that the needle extracts of P. densiflora have potential application as alternative anti-inflammatory agents or cosmetic material for skin health improvement.

The Effects of Bee Venom and Melittin Solution on PGE2, COX-2, and NF-kB Dependent Luciferase Activity in RAW 264.7 Cells (봉약침액(蜂藥鍼液)과 Melittin 약침액(藥鍼液)이 RAW 264.7 세포의 PGE2, COX-2 및 NF-kB에 미치는 영향(影響))

  • Jeong, Il-kook;Song, Ho-sueb
    • Journal of Acupuncture Research
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    • v.21 no.6
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    • pp.19-36
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    • 2004
  • Objective : The purpose of this study was to investigate the effect of Bee Venom and Melittin Solution on the lipopolysaccharide(LPS) and sodium nitroprusside(SNP)-induced expression of prostaglandin $E_2(PGE_2)$, cyclooxygenase-2(COX-2), nuclear factor kappa B($NF-{\kappa}B$) and nuclear factor kappa B($NF-{\kappa}B$) dependent luciferase activity in RAW 264.7 cells, a murine macrophage cell line. Methods : The expression of PGE2 was determined by determination of $PEG_2$, COX-2 was by western blotting with corresponding antibodies, $NF-{\kappa}B$ was by gel mobility shift assay method and $NF-{\kappa}B$ dependent luciferase activity was investigated by luciferase assay in RAW 264.7 cells. Results : 1. LPS and SNP-induced expression of $PEG_2$ was significant after 24hour. 2. The 0.5, 1 and $5{\mu}g/mL$ of bee venom and the 5 and $10{\mu}g/mL$ of melittin solution inhibited significantly LPS-induced expression of $PEG_2$ and, the $5{\mu}g/mL$ of bee venom and the 5 and $10{\mu}g/mL$ of melittin solution inhibited significantly SNP-induced expression of $PEG_2$ compared with control, respectively. The 0.5 and $1{\mu}g/mL$ of bee venom could not significantly inhibit SNP-induced expression of $PEG_2$ compared with control. 3. The $5{\mu}g/mL$ of bee venom and the 5 and $10{\mu}g/mL$ of melittin solution inhibited significantly LPS and SNP-induced expression of COX-2 compared with control, respectively. The 0.5 and $1{\mu}g/mL$ of bee venom inclined to decrease LPS and SNP-induced expression of COX-2 compared with control. 4. The 0.5, 1 and $5{\mu}g/mL$ of bee venom and the 5 and $10{\mu}g/mL$ of melittin solution inhibited significantly LPS and SNP-induced expression of $NF-{\kappa}B$ compared with control, respectively. 5. The 0.5, 1 and $5{\mu}g/mL$ of bee venom and the 5 and $10{\mu}g/mL$ of melittin solution inhibited significantly LPS-induced expression of $NF-{\kappa}B$ dependent luciferase activity and the 1 and $5{\mu}g/mL$ of bee venom and the 5 and $10{\mu}g/mL$ of melittin solution inhibited significantly SNP-induced expression of $NF-{\kappa}B$ dependent luciferase activity compared with control, respectively. The $NF-{\kappa}B$ inhibitor also inhibited significantly LPS and SNP-induced expression of $NF-{\kappa}B$ dependent luciferase activity compared with control. 6. The 0.5, 1 and $5{\mu}g/mL$ of bee venom and the 5 and $10{\mu}g/mL$ of melittin solution inhibited significantly LPS + IFN-${\gamma}$, TNF-${\alpha}$ and LPS + TNF-${\alpha}$-induced expression of $NF-{\kappa}B$ dependent luciferase activity compared with control, respectively. The $NF-{\kappa}B$ inhibitor also inhibited significantly LPS and SNP-induced expression of $NF-{\kappa}B$ dependent luciferase activity compared with control. Conclusions : These results suggest the inhibitory action of bee venom and melittin solution on the inflammatory mediators such as $PEG_2$, COX-2 and $NF-{\kappa}B$.

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Antibody Induced by the JY-Pol Pneumococcal Conjugate Protects Mice Against systemic Infection Due to Streptococcus pneumoniae (JY-Pol 접합백신으로 유도된 항페렴구균 항체의 보호효과)

  • Lee, Jue-Hee;Han, Yong-Moon
    • YAKHAK HOEJI
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    • v.48 no.6
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    • pp.369-373
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    • 2004
  • We previously reported that Streptococcus pneumoniae capsule attached to the surface protein (JY-Pol) was protective to systemic pneumococcal infection. The JY -Pol antigen induced IgM, IgG, and IgA in mice and provoked cell-mediated immunity. In this current study, we investigated the effect of anti JY-Pol antiserun and monoclonal antibody C2 (Mab C2) specific for the JY-Pol antigen against the pneumococcal disease. Mice that were given the antiserum survived longer than mice that received antiserum pre-absorbed with S.pneumoniae cells or DPBS as a negative control. Heat-treated anti JY-Pol antiserum resulted in survival rates similar to intact fresh JY-Pol antiserum. Mab C2 isolated from JY-Pol-immunized mice also enhanced resistance of naive mice against the pneumococcal diseaser. This protection by Mab C2 appeared to be mediated by opsonization as determined in a RAW 264.7 monocyte/macrophage cell line. Epitope analysis showed that Mab C2 epitope consisted of glucuronic acid and glucose that blocked the interaction of JY-Pol to the C2. Taken together, these data indicate that the antiserum induced by the JY-Pol, a naturally pneumococcal conjugate formula, mediated the protection by passive transfer, which was confirmed by protective effect of Mab C2.

Anti-cancer Activities of Ginseng Extract Fermented with Phellinus linteus

  • Lee, Jong-Jin;Kwon, Ho-Kyun;Jung, In-Ho;Cho, Yong-Baik;Kim, Kyu-Joong;Kim, Jong-Lae
    • Mycobiology
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    • v.37 no.1
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    • pp.21-27
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    • 2009
  • In the present study, the anti-cancer effects of ginseng fermented with Phellinus linteus (GFPL) extract were examined through in vitro and in vivo assays. GFPL was produced by co-cultivating ginseng and Phellinus linteus together. Ginsenoside Rg3, Rh1 and Rh2 are important mediators of anti-angiogenesis and their levels in GFPL were enriched 24, 19 and 16 times, respectively, more than that of ginseng itself through the fermentation. GFPL exhibited distinct anti-cancer effects, including growth inhibition of the human lung carcinoma cell line A549, and promotion of immune activation by stimulating nitric oxide (NO) production in Raw 264.7 cells. Further evidence supporting anti-cancer effects of GFPL was its significant prolongment of the survival of B16F10 cancer cell-implanted mice. These results suggest that the GFPL may be a candidate for cancer prevention and treatment through immune activation and anti-angiogenic effects by enriching Rg3, Rh1 and Rh2.

Evaluation of Antioxidant and Anti-Inflammatory Activities of Ascidian Tunic Carotenoids As a Source of Color Cosmetics (멍게껍질 카로테노이드의 색조 화장품 원료의 항산화, 항염증 기능성 평가)

  • Ticar, Bernadeth;Rohmah, Zuliyati;Bat-Erdene, Munkhjagal;Park, Si-Hyang;Choi, Byeong-Dae
    • KSBB Journal
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    • v.28 no.1
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    • pp.36-41
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    • 2013
  • Carotenoids are fat-soluble red-orange colored pigments found in plants and seafood-derived products, including algae, seaweeds, and fish muscle. In this study, we have demonstrated the molecular mechanism underlying the antioxidants and anti-inflammatory properties of ascidian tunic carotenoids using mouse macrophage cell line (RAW 264.7). Cell viability was not affected by treatment of carotenoids < 10 ${\mu}g/mL$. This treatment also showed negative inhibition on lipopolysaccharide (LPS)-stimulated nitric oxide (NO) and cyclooxygenase-2 (COX-2). The DPPH radical scavenging activity of carotenoids was 47.2% at 100 mg/mL. It also has a potential reducing power (1.025) comparable with ascorbic acid (1.584). The ascidian tunic carotenoids would make a candidate for the commercially interesting biologically active cosmetic pigments.