• Title/Summary/Keyword: Cyclooxygenase-2 promoter assay

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Comparison of In Vitro Antioxidant Activity and Cyclooxygenase-2 Promoter Inhibitory Activity in Harmonia axyridis Pallas and Coccinella septempunctata $Linn\dot{e}$ (약용곤충 무당벌레류 추출물의 항산화활성과 Cyclooxygenase-2 Promoter 억제활성 비교)

  • Heo Jin-Chul;Park Ja-Young;Hwang Jae-Sam;Park Hae-Cheol;Kang Seok-Woo;Hwang Seok-Jo;Yun Chi-Young;Kwon Taeg-Kyu;Lee Sang-Han
    • Food Science and Preservation
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    • v.13 no.4
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    • pp.513-518
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    • 2006
  • Insect resources have been widely recognized that seven millions of insects exhibit it own biological activity by whole body or its metabolic intermediates. In order to investigate antioxidant activity and compare the cyclooxygenase-2 promoter activity from insect extract, we tested in vitro antioxidant assays and cyclooxygenase-2 promoter assay in Coccinella septempunctata Linne and Harmonia axyridis extracts have the anti-oxidant and cyclooxygenase-2 inhibition activities, we examined the anti-oxidant assays including DPPH, FRAP and linoleic acid, ana inhibition of cyclooxygenase-2 expression using a cyclooxygenase-2 promoter-inserted stable cell line. We found that Harmonia axyridis Pallas extract had potentials to anti-oxidant activity and inhibited about 25% of cyclooxygenase-2 transcription activity. These findings indicate that Coccinella septempunctata Linne and Harmonia axyridis Pallas extracts could be an useful insect resource for agrobiotechnological purposes.

Comparison of Anti-Oxidative and Cox-2 Promoter Activities of Lepidoptera Extracts (Lepidoptera 추출물에 의한 항산화 및 Cox-2 프로모터 활성 비교)

  • Son, Hyeong-U;Heo, Jin-Chul;Lee, Sang-Han
    • Food Science and Preservation
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    • v.17 no.5
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    • pp.752-756
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    • 2010
  • Lepidoptera (butterflies) extracts, traditionally employed as medicines, have various biological activities. Five species of Lepidoptera (Papilio maackii, Papilio xuthus, Pieris rapae, Eurema hecabe, and Sasakia charonda) were extracted with distilled water (DW), dimethyl sulfoxide (DMSO), ethanol (EtOH), and methanol (MeOH). Each extract was analyzed for anti-oxidant and anti-inflammatory activities using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay method, the ferric reducing ability of plasma (FRAP) test, and a cyclooxygenase-2 (COX-2) promoter assay. The results suggest that Lepidoptera extracts have valuable anti-oxidant and anti-inflammatory properties, supporting the idea that the extracts may serve as a food biomaterial(s) preventing oxidative processes and inflammatory damage.

In Vitro Study of Anti-inflammatory and Analgesic Effects of Salvia Miltiorrhiza (SM) Extracts Using Luciferase Reporter Gene Assay (Luciferase reporter gene assay를 이용한 단삼(丹蔘)추출물의 소염, 진통작용에 대한 in vitro 연구)

  • Kim, Hee-Eun;Min, Sang-Yeon;Kim, Jang-Hyun
    • The Journal of Korean Medicine
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    • v.29 no.3
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    • pp.88-99
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    • 2008
  • Objectives: In order to identify the anti-inflammatory and analgesic properties of Salvia miltiorrhiza (Dan-Sam), widely used in Korean traditional medicine, an in vitro screening system was designed using pGL3, a luciferase reporter vector, and the tumor necrosis factor (TNF)-${\alpha}$ and cyclooxygenase (COX)-2 as target genes. Methods: The promoter regions of each gene were generated by PCR using the human chromosome as template DNA, and inserted into pGL3 vector with Kpn I and Hind III. The final construct was transfected into human myelomonocytic leukemia cells (U-937) that could be differentiated and activated by phorbol 12-myristate 13-acetate (PMA) or lipopolysaccharide (LPS). Using this system, the anti-inflammatory and analgesic effects of several herbal extracts regarded to have the medicinal effects of diminishing body heat and complementing Qi were tested. The chemicals PD98059 and berberine chloride were used as controls of the transcriptional inhibitors of TNF-${\alpha}$ and COX-2, respectively. Results: Salvia miltiorrhiza (Dan-Sam) demonstrated significant decrease of TNF-${\alpha}$ and COX-2 mRNA in the in vitro assay system. In MTT assay, Salvia miltiorrhiza (Dan-Sam) did not significantly inhibit the survival and proliferation of human myelomonocytic leukemia cells (U-937). Conclusions: Salvia miltiorrhiza (Dan-Sam) was found to exhibit the significant medicinal properties of anti-inflammatory and analgesic effects.

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Feedback Control of Cyclooxygenase-2 Expression by Prostaglandin E2 in Rheumatoid Synoviocytes

  • Min, So-Youn;Jung, Young Ok;Do, Ju-Ho;Kim, So-Yang;Kim, Jeong-Pyo;Cho, Chul-Soo;Kim, Wan-Uk
    • IMMUNE NETWORK
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    • v.3 no.3
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    • pp.201-210
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    • 2003
  • Objective: The role of prostaglandin $E_2$ (PGE2) in the etiopathogenesis of immune and inflammatory diseases has become the subject of recent debate. To determine the role of PGE2 in rheumatoid arthritis (RA), we tested the effect of exogenous PGE2 on the production of cyclooxygenase-2 (COX-2) by rheumatoid synoviocytes. Methods: Fibroblast-like synoviocytes (FLS) were prepared from the synovial tissues of RA patients, and cultured in the presence of PGE2. The COX-2 mRNA and protein expression levels were determined by RT-PCR and Western blot analysis, respectively. The PGE2 receptor subtypes in the FLS were analyzed by RT-PCR. Electrophoretic mobility shift assay (EMSA) was used to measure the NF-${\kappa}B$ binding activity for COX-2 transcription. The in vivoeffect of PGE2 on the development of arthritis was also tested in collagen induced arthritis (CIA) animals. Results: PGE2 ($10^{-11}$ to $10^{-5}M$) dose-dependently inhibited the expression of COX-2 mRNA and the COX-2 protein stimulated with IL-$1{\beta}$, but not COX-1 mRNA. NS-398, a selective COX-2 inhibitor, displayed an additive effect on PGE2-induced COX-2 downregulation. The FLS predominantly expressed the PGE2 receptor (EP) 2 and EP4, which mediated the COX-2 suppression by PGE2. Treatment with anti-IL-10 monoclonal antibodies partially reversed the PGE2-induced suppression of COX-2 mRNA, suggesting that IL-10 may be involved in modulating COX-2 by PGE2. Experiments using an inducer and an inhibitor of cyclic AMP (cAMP) suggest that cAMP is the major intracellular signal that mediates the regulatory effect of PGE2 on COX-2 expression. EMSA revealed that PGE2 inhibited the binding of NF-${\kappa}B$ in the COX-2 promoter via a cAMP dependent pathway. In addition, a subcutaneous injection of PGE2 twice daily for 2 weeks significantly reduced the incidence and severity of CIA as well as the production of IgG antibodies to type II collagen. Conclusion: Our data suggest that overproduced PGE2 in the RA joints may function as an autocrine regulator of its own synthesis by inhibiting COX-2 production and may, in part, play an anti-inflammatory role in the arthritic joints.

Effect of ω3-Fatty Acid Desaturase Gene Expression on Invasion and Tumorigenicity in Human Tongue Squamous Cell Carcinoma Cells (인체 혀의 편평세포암 세포에서 ω3-fatty acid desaturase 유전자 발현이 침윤 및 종양형성에 미치는 영향)

  • Hong, Tae-Hwa;Shin, Soyeon;Han, Seung-Hyeon;Hwang, Byung-Doo;Lim, Kyu
    • Journal of Life Science
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    • v.28 no.8
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    • pp.945-954
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    • 2018
  • Omega-3 polyunsaturated fatty acids (${\omega}3$-fatty acid) have been found to possess anticancer properties in a variety of cancer cell lines and animal models, but their effects in human tongue squamous cell carcinomas (SCCs) remain unclear. This study was designed to examine the effect of ${\omega}3$-fatty acid desaturase (fat-1) gene expression on invasion and tumorigenicity in human tongue SCC cells and the molecular mechanism of its action. Docosahexaenoic acid (DHA) treatment inhibited in vitro invasion in a dose-dependent manner. In zymography, matrix metalloproteinase-9 (MMP-9) and Matrix metallopeptidase-2 (MMP-2) activities were reduced, and MMP-9 and MMP-2 promoter activities were inhibited by the DHA treatment. In addition, cyclooxygenase-2 (COX-2) and vascular endothelial growth factor (VEGF) promoter reporter activities were inhibited in SCC-4 and SCC-9 cells after the DHA treatment. To investigate the effect of a high level of endogenous ${\omega}3$ fatty acids, a stable SCC-9 cell line expressing the ${\omega}3$-desaturase gene (fSCC-9sc) was generated. The growth rate and colony-forming capacity of fSCC-9sc were remarkably decreased as compared with those of fSCC-9cc. Likewise, the tumor size and volume of fSCC-9sc implanted into nude mice were significantly inhibited, with increases in the cell death index. Furthermore, a transwell chamber invasion assay showed a reduction in cell invasion of the fSCC-9sc lines when compared with that of the fSCC-9cc line. These findings suggested that fat-1 gene expression inhibited tumorigenicity, as well as invasion in human tongue SCC cells. Thus, utilization of ${\omega}3$ fatty acids may represent a promising therapeutic approach for chemoprevention and the treatment of human tongue SCCs.

Comparison of Antioxidant, Anticancer and Immunomodulating Activities of Extracts from DongChongXiaCao (동충하초의 균사 및 배양액의 항산화, 항암, 면역활성의 비교)

  • Heo, Jin-Chul;Nam, Sung-Hee;Kang, Seok-Woo;Hong, In-Pyo;Lee, Kwang-Kil;Park, Ja-Young;Kim, Kyung-Hae;Han, Song-Yi;Lee, Sang-Han
    • Food Science and Preservation
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    • v.14 no.6
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    • pp.681-687
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    • 2007
  • Biological properties of DongChongXiaCao extracts and culture supernatants were evaluated using DPPH and FRAP (antioxidants), Raw 264.7 (NO production), B16-F1 cells (cell migration activity) and HUVECs (angiogenesis activity). We found that antioxidant activity was higher in mycelium culture supernatants than in mycelial extracts. Mycelial extracts and culture supernatants inhibited or increased cyclooxygenase-2 transcription activity and NO production. Various extracts and culture supernatants inhibited B16 cell migration and motility, and inhibited HUVEC tube formation. These findings indicate that DongChongXiaCao extracts and products of mycelium could be a useful biological resource for anti-oxidant and anti-cancer purposes.