• Title/Summary/Keyword: cytotoxicity assay

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Involvement of Early Growth Response Gene 1 (EGR-1) in Growth Suppression of the Human Colonic Tumor Cells By Apigenin and Its Derivative Isovitexin (Apigenin과 대사물 isovitexin에 의한 인체 대장암세포의 세포활성 억제효과에 있어서의 EGR-1의 역할 연구)

  • Moon, Yu-Seok;Cui, Lei-Guang;Yang, Hyun
    • Journal of Life Science
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    • v.17 no.1 s.81
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    • pp.110-115
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    • 2007
  • It has been previously described that transcription factor early growth response gene product 1 (EGR-1) functions as a tumor suppressor gene. This study was conducted to demonstrate that EGR-1 induction by phytochemical apigenin and its derivative isovitexin can mediate the growth suppression of the intestinal epithelial tumor cells. Apigenin and isovitexin induced EGR-1 gene expression both in the dose and time-dependent manners. Moreover the induction was relatively late around 9-12 hr after treatment of HCT-116 cells, while several anti-inflammatory agent such as NSAIDS and catechins elicit the ECR-1 gene expression at much earlier time about 1-3 hr after treatment. In terms of signal transduction, ERK1/2 was critical for apigenin-induced EGR-1 gene expression and its promoter activation. When EGR-1 gene expression was blocked with EGR-1 small interference RNA, the cytotoxicity of apigenin in the human epithelial cells was attenuated, suggesting the involvement of EGR-1 in the anti-tumoric activity of apigenin. To link the EGR-1 induction to EGR-1-regulated gene products in colon cancer, NSAID-Activated Gene 1 (NAG-1) was demonstrated to be elevated by apigenin and isovitexin at 24-48 hr after treatment. Taken together, apigenin-activated ERK1/2 mediated EGR-1 gene induction, which was associated with suppression of the cellular viability by apigenin compound.

Antiobesity Activity of Chrysanthemum zawadskii Methanol Extract (구절초 추출물의 항비만 활성)

  • Park, Jung Ae;Jin, Kyong-Suk;Kwon, Hyun Ju;Kim, Byung Woo
    • Journal of Life Science
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    • v.25 no.3
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    • pp.299-306
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    • 2015
  • Chrysanthemum zawadskii, a herbaceous perennial plant belonging to the Compositae, grows wild in Asian countries, including Japan, China, and Korea. The biological, antioxidative, anti-inflammatory, and antibacterial activities of C. zawadskii have been reported, its antiobesity activity has not been elucidated. In the present study, the effect of C. zawadskii methanol extract (CZME) on pancreatic lipase enzyme activity, adipocyte differentiation, and adipogenesis was investigated using an in vitro assay and a cell model system. CZME effectively suppressed lipase enzyme activity in a dose-dependent manner. CZME also inhibited insulin, dexamethasone, 3-isobutyl-1-methylxanthine (MDI)-induced adipocyte differentiation, lipid accumulation, and the level of triglyceride in 3T3-L1 preadipocytes in a dose-dependent manner, without cytotoxicity. The antiobesity effect of CZME might be modulated by gene and protein expression of cytidine-cytidine-adenosine-adenosine-thymidine (CCAAT)/enhancer binding proteins (C/EBP) α, C/EBPβ, and the peroxisome proliferator-activated receptor γ (PPAR γ). CZME also triggered lipolysis in a dose-dependent manner in MDI-induced 3T3-L1 preadipocytes. Taken together, these results provide important new insights into the antiobesity activities of C. zawadskii, showing that they involve pancreatic lipase inhibition, as well as antiadipogenic and lipolysis effects. CZME might be a promising source in the field of nutraceuticals. However, the active compounds that confer the antiobesity activities of CZME need to be identified.

Anti-Inflammatory Effect of Sedum takesimense Nakai Water Extract in RAW 264.7 Cells (섬기린초 물 추출물의 마우스 대식세포에서 항염증 효능)

  • Jang, Ji Hun;Jung, Ho Kyung;Ko, Jae Hyung;Sim, Mi Ok;Woo, Kyeong Wan;Kim, Tae Muk;Lee, Ki Ho;Ahn, Byeong Kwan;Cho, Hyun Woo;Cho, Jung Hee;Jung, Won Seok
    • Korean Journal of Medicinal Crop Science
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    • v.24 no.3
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    • pp.228-236
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    • 2016
  • Background: Sedum takesimense Nakai has been used as folk medicine in Korea. The present study aimed to determine the biological activity of S. takesimense by investigating the anti-inflammatory effects of S. takesimense water extract (SKLC) on the lipopolysaccharide-induced inflammatory response in RAW 264.7 cells. Methods and Results: Cytotoxicity of SKLC on RAW 264.7 cells was determinded by performing MTS assay was found to have no cytotoxic effect on RAW 264.7 cells at a concentration range of 62500μg/m. Further, pretreatment of SKLC inhibited lipopolysaccharide-induced nitric oxide (NO) production in a dose-dependent manner. To determined the inhibitory mechanisms of SKLC on inflammatory mediators, we assessed the inducible nitric oxide synthase (iNOS) and cyclooxygnease-2 (COX-2) pathways. The activities of these pathways were decreased in a dose-dependent manner by SKLC. The production of tumor necrosis factor-α (TNF-α), interleukin (IL)1β, and IL-6 were also reduced. Conclusions: These results suggest that the down regulation of iNOS, COX-2, TNF-α, IL-1β, and IL-6 expression by SKLC are mediated by the down regulation of nuclear factor-κB (NF-κB) activity, a transcription factor necessary for pro-inflammatory mediators. This might be the mechanism underlying the anti-inflammatory effects of SKLC.

Effect of Piryongbanggamgil-tang on Airway Mucin Secretion, Production, Gene Expression and Hypersecretion of Mucus (필용방감길탕이 기도 뮤신의 분비, 생성, 유전자 발현 및 점액 과다 분비에 미치는 영향)

  • Kim, Yoon Young;Min, Sang Yeon;Kim, Jang Hyun
    • The Journal of Pediatrics of Korean Medicine
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    • v.28 no.2
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    • pp.56-71
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    • 2014
  • Objectives In this study, the author tried to investigate whether piryongbang-gamgil-tang (PGGT) significantly affect in vitro airway mucin secretion, PMA- or EGF- or TNF-α-induced MUC5AC mucin production / gene expression from human airway epithelial cells and increase in airway epithelial mucosubstances and hyperplasia of tracheal goblet cells of rats. Materials and Methods For in vitro experiment, confluent RTSE cells were chased for 30 min in the presence of PGGT to assess the effect of PGGT on mucin secretion by enzyme-linked immunosorbent assay (ELISA). Also, effect of PGGT on PMA- or EGFor TNF-α-induced MUC5AC mucin production and gene expression from human airway epithelial cells (NCI-H292) were investigated. Confluent NCI-H292 cells were pretreated for 30 min in the presence of PGGT and treated with PMA (10 ng/ml) or EGF (25 ng/ml) or TNF-α (0.2 nM) for 24 hrs, to assess both effect of PGGT on PMA- or EGF- or TNF-α-induced MUC5AC mucin production by ELISA and gene expression by reverse transcription-polymerase chain reaction (RT-PCR). For in vivo experiment, the author induced hypersecretion of airway mucus and goblet cell hyperplasia by exposure of rats to SO2 during 3 weeks. Effect of orally-administered PGGT during 2 weeks on increase in airway epithelial mucosubstances from tracheal goblet cells of rats and hyperplasia of goblet cells were assesed by using histopathological analysis after staining the epithelial tissue with alcian blue. Possible cytotoxicities of PGGT in vitro were assessed by examining LDH release from RTSE cells and the rate of survival and proliferation of NCI-H292 cells. In vivo liver and kidney toxicities of PGGT were evaluated by measuring serum GOT/GPT activities and serum BUN/creatinine concentrations of rats after administering PGGT orally. Results (1) PGGT did not affect in vitro mucin secretion from cultured RTSE cells. (2) PGGT significantly inhibited PMA-, EGF-, and TNF-α-induced MUC5AC mucin productions and the expression levels of MUC5AC mRNA from NCI-H292 cells. (3) PGGT decreased the amount of intraepithelial mucosubstances and showed the tendency of expectorating airway mucus already produced. (4) PGGT increased LDH release from RTSE cells. However, PGGT did not show in vivo liver and kidney toxicities and cytotoxicity to NCI-H292 cells. Conclusion The result from this study suggests that PGGT can regulate the production and gene expression of airway mucin observed in diverse respiratory diseases accompanied by mucus hypersecretion and do not show in vivo toxicity to liver and kidney functions after oral administration. Effect of PGGT with their components should be further studied using animal experimental models that reflect the diverse pathophysiology of respiratory diseases through future investigations.

Physiological Activities using Root and Stem Extracts of Cymbidium (심비디움 뿌리 및 줄기 추출물의 생리 활성)

  • Kim, Hye-Ran;Park, Gyu-Nam;Jung, Bo-Kyoung;Shin, Yu-Su;Chang, Kyung-Soo
    • Journal of the Korean Applied Science and Technology
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    • v.33 no.4
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    • pp.848-854
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    • 2016
  • Cymbidium is one of perennial herbs belonging to the Orchidaceae and is known as a medicinal plant. However, its scientific data are insufficient. The purpose of this study is to extract from root and stem of Cymbidium, to investigate the biological effects of them. Cymbidium antibacterial effects of the extracts were performed by antibacterial test against Staphylococcus aureus (S. aureus), Staphylococcus saphrophyticus (S. saprophyticus), Proteus vulgaris (P. vulgaris) and Klebsiella pneumonia (K. pneumonia). Antioxidant effects of the extracts were carried out by DPPH radical scavenging. Total phenolic contents were also determined. Moreover, Cell viability of extracts against MTT assay was cell viability against HepG2 cell and also measures Cholesterol adsorptivity of extracts. In this study, the extracts inhibited the growth of bacteria. Particularly Cymbidium root extracts by only ethanol extraction showed highest antimicrobial effect against S. aureus. The Cymbidium stem extracts by both ethanol extraction and sonication for 1 hour had higher antioxidant activites as well as total phenolic contents. Cell cytotoxicity showed higher than 50μg/mL. Cholesterol adsorptivity showed lower than 20%. These results suggest that the Cymbidium might be a source of anti-bacterials and anti-oxidants.

Whitening Effect of Dayflower (Commelina communis L.) Extract by Inhibition of N-Linked Glycosylation Process and Melanogenesis (N-Linked Glycosylation 저해에 의한 닭의장풀 추출물의 미백효능)

  • Park, Sun-Hee;Lee, Bang-Yong;Lee, Seung-Hyun;Han, Chang-Sung;Kim, Jin-Guk;Kim, Kyoung-Tae;Kim, Ki-Ho;Kim, Young-Heui
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.35 no.1
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    • pp.73-78
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    • 2009
  • In order to investigate the potential of a Dayflower (Commelina communis L.) extract as an active in gredient for whitening cosmetics, we prepared aqueous Commelina communis L. extract We measured its mushroom tyrosinase inhibitory activity, cellular tyrosinase activity, and melanin synthesis inhibitory activity in B16 melanoma cells. It did not show inhibitory activity against mushroom tyrosinase but showed melanin synthesis inhibitory activity. In a melanin synthesis inhibition assay using mouse B16-F10 melanoma cell, it suppressed melanin production up to 32% at a concentration of 1,000μ/mL without cytotoxicity, and also reduced cellular tyrosinase activity to above 50 % above the concentration of 250μg/mL. In study on the melanogenic protein expressions, it had especially influence on expression of tyrosinase protein, which is a well-known key protein on melanogenesis, and tyrosinase expression was gradually decreased in a dose-dependent. Dayflower also blocked N-glycosylation of TRP-2, but affected on the expression of TRP-1 rather than on blocking of N-glycosylation processing. Therefore, this result suggests that aqueous Commelina communis L. extract could be used as an active ingredient for whitening cosmetics.

The Coffee Sliver Skin Extracts from Coffee Beans Exhibited Cosmetic Properties with Antioxiant Activity and Inhibitory Effects for Elastase, Collagenase and Tyrosinase (커피 은피 추출물의 항산화 효과와 엘라스타제, 콜라게나제 및 티로시나제 저해효과)

  • Lee, Kyung Eun;Son, Sang Hyeok;Kang, Sang Gu
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.44 no.1
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    • pp.39-48
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    • 2018
  • The coffee silver skin is a part of coffee beans. We report that the coffee sliver skin extracts exhibited cosmetic properties of antioxidant, anti-winkle and whitening effects. The ethanol extracts of silver skin showed free radical scavenging activity up to 92.26% in 50μg/mL, especially against DPPH radical and ABTS radical cation. The silver skin extracts showed inhibitory effects for tyrosinase activity and DOPA oxidation in a dose-dependent manner, suggesting the extracts retain for the whitening property in cosmetics. The coffee silver skin extracts effectively inhibited the elastase and collagenase. Cytotoxicity of the coffee silver skin extracts was measured by the colorimetric MTS assay. The viability of the human keratinocytes (HaCaT) treated with the coffee silver skin extracts was same as that of untreated cells, indicating the extracts are safe to human cells. Here, we suggest that the silver skin extracts of coffee bean could be a potential natural substance for anti-winkle, whitening, antioxidant properties for cosmetics.

Invitro and Virtual Screening of Bioactive Molecule from Mycelium of Trichoderma atroviride Inhibit the UDP-3-O-(R-3-hydroxymyristoyl)-N-acetylglucosamine Deacetylases (LpxC) for Treatment of Bacterial Infection

  • Saravanakumar, Kandasamy;Park, Cheol-Ho;Wang, Myeong-Hyeon
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.67-67
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    • 2018
  • Trichoderma species are a rich source of metabolites, but less known for biomedical potential. This work deals with antibacterial and antioxidant potentials of intracellular non-cytotoxic metabolites, extracted from Trichoderma atroviride (KNUP001). A total of 53 fractions was collected by column chromatography and tested for cytotoxicity by MTT assay. Only one fraction (F41) was found to be non-toxic to Vero cells with 95.4±0.61 of survival. The F41 was then subjected to chemical analysis, antibacterial and antioxidant assays. The F41 at 500μg.ml1 showed the total antioxidant of 48.70±2.90, DPPH radical scavenging activity of 37.25±2.25, nitric oxide (NO) radical scavenging activity of 54.55±1.95 and H2O2 radical scavenging activity of 43.75±3.21. The F41 at 25μg.ml1 displayed antibacterial activity against E. coli (14.25±0.2mm), P. mirabilis (10.4±0.6mm), S. dysenteriae (18.6±03mm), S. paratyphi A (14.1±1.1mm), E. aerogenes (5.6±0.4mm) and S. marcescens (14.25±0.2mm). GC-MS analysis revealed the dominant presence of oleic acid C 18.1 (63.18%), n-hexadecanoic acid (6.17%), and ethyl oleate (4.93%) and potent molecules such as 8-[(2E)-2-(3-hydroxybenzylidene)hydrazinyl]-1,3,7-trimethyl-3,7-dihydro-1H-purine-2,6-dione, 2-(Dimethylamino)ethyl (1Z)-N-hydroxy-2-(4-morpholinyl)-2-oxoethanimidothioate, Fluorene in the F41, and virtual study revealed that these molecules are likely responsible for the antibacterial activities of F41. Hence, further investigation deserves on purification and characterization of the active metabolites from T. atroviride strain KNUP001 towards developing molecular leads to effective antibacterial drugs, and non-toxic to host cells.

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Mechanism underlying NO-induced apoptosis in human gingival fibroblasts

  • Hwang, In-Nam;Jeong, Yeon-Jin;Jung, Ji-Yeon;Lee, Jin-Ha;Kim, Kang-Moon;Kim, Won-Jae
    • International Journal of Oral Biology
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    • v.34 no.1
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    • pp.7-14
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    • 2009
  • Nitric oxide (NO) acts as an intracellular messenger at the physiological level but can be cytotoxic at high concentrations. The cells within periodontal tissues, such as gingival and periodontal fibroblasts, contain nitric oxide syntheses and produce high concentrations of NO when exposed to bacterial lipopolysaccharides and cytokines. However, the cellular mechanisms underlying NO-induced cytotoxicity in periodontal tissues are unclear at present. In our current study, we examined the NO-induced cytotoxic mechanisms in human gingival fibroblasts (HGF). Cell viability and the levels of reactive oxygen species (ROS) were determined using a MTT assay and a fluorescent spectrometer, respectively. The morphological changes in the cells were examined by Diff-Quick staining. Expression of the Bcl-2 family and Fas was determined by RT-PCR or western blotting. The activity of caspase-3, -8 and -9 was assessed using a spectrophotometer. Sodium nitroprusside (SNP), a NO donor, decreased the cell viability of the HGF cells in a dose- and time-dependent manner. SNP enhanced the production of ROS, which was ameliorated by NAC, a free radical scavenger. ODQ, a soluble guanylate cyclase inhibitor, did not block the SNP-induced decrease in cell viability. SNP also caused apoptotic morphological changes, including cell shrinkage, chromatin condensation, and DNA fragmentation. The expression of Bax, a member of the proapoptotic Bcl-2 family, was upregulated in the SNP-treated HGF cells, whereas the expression of Bcl-2, a member of the anti-apoptotic Bcl-2 family, was downregulated. SNP augmented the release of cytochrome c from the mitochondria into the cytosol and enhanced the activity of caspase-8, -9, and -3. SNP also upregulated Fas, a component of the death receptor assembly. These results suggest that NO induces apoptosis in human gingival fibroblast via ROS and the Bcl-2 family through both mitochondrial- and death receptor-mediated pathways. Our data also indicate that the cyclic GMP pathway is not involved in NO-induced apoptosis.

Antimutagenic and Cytotoxic Effects of Fagopyrum esculentum Moenech Noodles Extracts (메밀 국수 추출물의 항돌연변이원성 및 세포독성 효과)

  • Yoo, Kwang-Ha;Kim, Soo-Hyun;Ham, Young-An;Yoo, Soo-Jung;Oh, Hyun-Taek;Ham, Seung-Shi
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.10
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    • pp.1291-1296
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    • 2006
  • This study was performed to determine the antimutagenic and anticancer effects of Fagopyrum esculentum Moenech noodles (FEMN) extracts using Ames test and cytotoxicity, respectively. FEMN made buckwheat wet noodles (BWN), buckwheat extruded noodles (BEN) and buckwheat dehydrated noodles (BDN) by 60% buckwheat flour and 70% buckwheat flour. The inhibitory effects of FEMN extracts on cell proliferation in A549, Hep3B, MCF-7, AGS and HeLa were investigated by SRB assay. The cytotoxic effects of FEMN against the cell lines with human lung carcinoma (A549), human gastric carcinoma (AGS), human hepatocellular carcinoma (Hep3B), human cervical adenocarcinoma (HeLa) and human breast adenocarcinoma (MCF-7) were inhibited with the increase of the extract concentration. The treatment of 1.0 mg/mL FEMN of 60% BEN extracts showed strong cytotoxicities of 74.7%, 75.3% and 70.5% against AGS, A549 and HeLa, respectively. The inhibition rate of 70% BWN of FEMN extracts in the S. Typhimurium TA100 strain showed 41% against the mutagenesis induced by MNNG. The inhibition rate of 70% BEN of FEMN extracts in the S. Typhimurium TA98 strain showed 45% against the mutagenesis induced by 4NQO.