• Title/Summary/Keyword: in-cell western

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Anti-inflammatory Activity of Antimicrobial Peptide Zophobacin 1 Derived from the Zophobas atratus (아메리카왕거저리 유래 항균 펩타이드 조포바신 1의 항염증활성)

  • Shin, Yong Pyo;Lee, Joon Ha;Kim, In-Woo;Seo, Minchul;Kim, Mi-Ae;Lee, Hwa Jeong;Baek, Minhee;Kim, Seong Hyun;Hwang, Jae Sam
    • Journal of Life Science
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    • v.30 no.9
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    • pp.804-812
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    • 2020
  • The giant mealworm beetle, Zophobas atratus (Coleoptera: Tenebrionidae) has been used as a protein source for small pets and mammals. Recently, it was temporarily registered in the list of the Food Code. We previously performed an in silico analysis of the Zophobas atratus transcriptome to identify putative antimicrobial peptides and identified several antimicrobial peptide candidates. Among them, we assessed the antimicrobial and anti-inflammatory activities of zophobacin 1 that was selected bio-informatically based on its physicochemical properties against microorganisms and mouse macrophage Raw264.7 cells. Zophobacin 1 showed antimicrobial activities against microorganisms without inducing hemolysis and decreased the nitric oxide production of the lipopolysaccharide-induced Raw264.7 cells. Moreover, ELISA and Western blot analysis revealed that zophobacin 1 reduced expression levels of pro-inflammatory enzymes such as inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). We also investigated expression of pro-inflammatory cytokines (interleukin-6 and interleukin-1β) production through quantitative real time-PCR and ELISA. Zophobacin 1 markedly reduced the expression level of cytokines through the regulation of mitogen-activated protein kinases (MAPKs) and nuclear factor kappa B (NF-κB) signaling. We confirmed that zophobacin 1 bound to bacterial cell membranes via a specific interaction with lipopolysaccharides. These data suggest that zophobacin 1 could be promising molecules for development as antimicrobial and anti-inflammatory therapeutic agents.

Protective effect of Gabjubaekmok (Diospyros kaki) extract against amyloid beta (Aβ)-induced cognitive impairment in a mouse model (아밀로이드 베타(amyloid beta)로 유도된 인지장애 마우스 모델에서 갑주백목(Diospyros kaki) 추출물의 인지기능 및 뇌 신경세포 보호 효과)

  • Yoo, Seul Ki;Kim, Jong Min;Park, Seon Kyeong;Kang, Jin Yong;Han, Hye Ju;Park, Hyo Won;Kim, Chul-Woo;Lee, Uk;Heo, Ho Jin
    • Korean Journal of Food Science and Technology
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    • v.51 no.4
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    • pp.379-392
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    • 2019
  • The current study investigated the effect of Gabjubaekmok (Diospyros kaki) ethanolic extract (GEE) on $H_2O_2$-induced human neuroblastoma MC-IXC cells and amyloid beta $(A{\beta})_{1-42}$-induced ICR (Institute of Cancer Research) mice. GEE showed significant antioxidant activity that was evaluated based on ABTS, DPPH scavenging activity, and inhibition of malondialdehyde (MDA) and acetylcholinesterase activity. Further, GEE inhibited ROS production and increased cell viability in $H_2O_2$-induced MC-IXC cells. Administration of GEE ameliorated the cognitive dysfunction on $A{\beta}$-induced ICR mice as evaluated using Y-maze, passive avoidance, and Morris water maze tests. Results of ex vivo test using brain tissues showed that, GEE protected the cholinergic system and mitochondrial functions by increasing the levels of antioxidants such as ROS, mitochondrial membrane potential (MMP), and adenosine triphosphate (ATP) against $A{\beta}$-induced cognitive dysfunction. Moreover, GEE decreasd the expression levels of apoptosis-related proteins such as $TNF-{\alpha}$, p-JNK, p-tau, BAX and caspase 3. While, expression levels of p-Akt and $p-GSK3{\beta}$ increased than $A{\beta}$ group. Finally, gallic acid was identified as the main compound of GEE using high performance liquid chromatography.

Anti-wrinkle and Whitening Effects of Essential Oil from Abies koreana (Abies koreana 유래 정유의 항주름 및 미백 효과)

  • Song, Byeong-Wook;Song, Min-Jeong;Park, Mi-Jin;Choi, Don-Ha;Lee, Sung-Suk;Kim, Myungkil;Hwang, Ki-Chul;Kim, Il-Kwon
    • Journal of Life Science
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    • v.28 no.5
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    • pp.524-531
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    • 2018
  • The essential oil from Abies koreana E.H. Wilson had been developed, however, its efficacy has not yet been studied especially in terms of skin care research. The aim of this study is to investigate the effects of Abies koreana extracts (AKE) on melanogenesis and wrinkle formation in B16F10 melanoma cells (B16F10) and human dermal fibroblast cell line (HDF). The essential oil was extracted by hydrodistillation method and purified by anhydrous sodium sulfate. At a concentration of $10^{-5}$-fold, viability in these cells had been defined by cytotoxicity assays. Anti-melanogenic effects on B16F10 were evaluated using tyrosinase inhibition assay, and real-time PCR for verifying gene expression of tyrosinase, tyrosinase related protein-1 and -2 (TRP-1 and -2). AKEs reduced about 5-fold of tyrosinase inhibitory activity compared to ${\alpha}$-melanocyte-stimulating hormone (${\alpha}$-MSH)-induced group and about 30% reduction compared to Arbutin induced group. The mRNA levels of three melanin-related factors were increased, separately. To investigate the effects of anti-wrinkle, procollagen type I c peptide synthesis assay (PIP) and Western blot were performed. At AKE-treated group, PIP was up-regulated and the expression of collagen type 1 and matrix metalloproteinase (MMP)-1 were improved. Furthermore, AKE presented anti-wrinkle effects by increasing UVB-inhibited collagen type 1 expression, and reducing UVB-induced MMP-1 production at $60mJ/cm^2$ of UVB radiation. Therefore, Abies koreana extracts has potentials as a safe and an effective skin ingredient for whitening and anti-wrinkle.

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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Effects of 2-methoxy-1,4-naphthoquinone (MQ) on MCP-1 Induced THP-1 Migration (MCP-1에 의해 유도된 THP-1 유주에 미치는 2-methoxy-1,4-naphthoquinone (MQ)의 영향)

  • Kim, Si Hyun;Park, Bo Bin;Hong, Sung Eun;Ryu, Sung Ryul;Lee, Jang Ho;Kim, Sa Hyun;Lee, Pyeongjae;Cho, Eun-Kyung;Moon, Cheol
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.2
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    • pp.245-251
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    • 2019
  • This study examined the effects of 2-methoxy-1,4-naphthoquinone (MQ) on the monocyte chemoattractant protein-1 (MCP-1)-induced migration of monocytes, which is an important phenomenon for the body defense and immune response. MQ is a major component extracted from Impatiens balsamina leaves, which have been used for many years in Asian medicine for the treatment of a range of diseases and pain. The cytotoxicity of MQ began to appear at a concentration of $10{\mu}M$, and approximately 50% cytotoxicity was confirmed at $100{\mu}M$. The MCP-1 induced migration of the THP-1 monocyte cell line increased after MQ treatment in a dose dependent manner and the largest increase was observed at $0.1{\mu}M$. The level of cAMP expression decreased after a treatment with $0.1{\mu}M$ MQ. The phosphorylation of extracellular signal-regulated kinases 1/2 (Erk1/2), a key signaling protein involved in the signaling pathway of C-C motif chemokine receptor 2 (CCR2), a receptor for MCP-1, was increased by the simultaneous treatment of $0.1{\mu}M$ MQ. These results show that MQ increases the MCP-1-induced migration of THP-1, decreases the level of cAMP expression, and increases the level of Erk1/2 phosphorylation.

Anti-inflammatory effect in macrophages according to the mixing ratio of acemannan and aloesin (Acemannan과 aloesin의 혼합 비율에 따른 대식세포에서의 항염증 효과)

  • Hyo-Min Kim;Jeong-Hwan Kim;Dan-Hee Yoo;Se-Yeong Jeon;Hyun-Jin Kim;Seon-Gil Do;In-Chul Lee;Jung-Wook Kang
    • Food Science and Preservation
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    • v.31 no.2
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    • pp.315-323
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    • 2024
  • This study aims to confirm the anti-inflammatory activities of acemannan and aloesin, which have been studied for various efficacies at various mixed sample ratios. The mixed samples were mixed at a ratio of 1:1 (AA-1), 1:2 (AA-2), 1:3 (AA-3), 2:1 (AA-4), and 3:1 (AA-5). Seven samples were evaluated for their cytotoxic ability on macrophages, and the results showed that all cell viability was over 90% at a concentration of 100 ㎍/mL. First, due to the NO production inhibitory activity, a better inhibitory effect was achieved when using a mixed sample rather than a single material. Afterward, the activity of inhibiting the production of PGE2, TNF-α, and IL-6 was confirmed using a mixed sample. It was confirmed that AA-2 had the best inhibitory activity on producing PGE2, TNF-α, and IL-6 rather than AA-1, AA-3, AA-4, and AA-5. For this reason, experiments were conducted using AA-2 to determine the protein expression levels of iNOS and COX-2, which are inflammation-related proteins. It was confirmed that AA-2 inhibited iNOS and COX-2 protein expression by 25.01% and 27.27%, respectively, compared to the LPS-alone treatment group. In conclusion, the mixed sample of acemannan and aloesin is judged to have anti-inflammatory activity and can potentially to be used as a functional material.

Acoustic images of the submarine fan system of the northern Kumano Basin obtained during the experimental dives of the Deep Sea AUV URASHIMA (심해 자율무인잠수정 우라시마의 잠항시험에서 취득된 북 구마노 분지 해저 선상지 시스템의 음향 영상)

  • Kasaya, Takafumi;Kanamatsu, Toshiya;Sawa, Takao;Kinosita, Masataka;Tukioka, Satoshi;Yamamoto, Fujio
    • Geophysics and Geophysical Exploration
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    • v.14 no.1
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    • pp.80-87
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    • 2011
  • Autonomous underwater vehicles (AUVs) present the important advantage of being able to approach the seafloor more closely than surface vessel surveys can. To collect bathymetric data, bottom material information, and sub-surface images, multibeam echosounder, sidescan sonar (SSS) and subbottom profiler (SBP) equipment mounted on an AUV are powerful tools. The 3000m class AUV URASHIMA was developed by the Japan Agency for Marine-Earth Science and Technology (JAMSTEC). After finishing the engineering development and examination phase of a fuel-cell system used for the vehicle's power supply system, a renovated lithium-ion battery power system was installed in URASHIMA. The AUV was redeployed from its prior engineering tasks to scientific use. Various scientific instruments were loaded on the vehicle, and experimental dives for science-oriented missions conducted from 2006. During the experimental cruise of 2007, high-resolution acoustic images were obtained by SSS and SBP on the URASHIMA around the northern Kumano Basin off Japan's Kii Peninsula. The map of backscatter intensity data revealed many debris objects, and SBP images revealed the subsurface structure around the north-eastern end of our study area. These features suggest a structure related to the formation of the latest submarine fan. However, a strong reflection layer exists below ~20 ms below the seafloor in the south-western area, which we interpret as a denudation feature, now covered with younger surface sediments. We continue to improve the vehicle's performance, and expect that many fruitful results will be obtained using URASHIMA.

Whitening Activity of Sambucus Sieboldiana Var. Pendula (Nakai) Extract (말오줌나무 추출물의 미백활성 검증)

  • Yoo, Dan-Hee;Kim, Jin-Tae;Oh, Min-Jeong;Yeom, Hyeon-Ji;Lee, Jin-Young
    • Journal of Life Science
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    • v.29 no.3
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    • pp.279-286
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    • 2019
  • This study evaluated the anti-oxidant and whitening effects of a 70% ethanol extract of the Sambucus sieboldiana var. pendula (Nakai) (SS). At $1,000{\mu}g/ml$ concentration, the electron donating ability of this SS extract was found to be 86.21% and the ABTS+ radical scavenging ability was 97.9%. In terms of whitening activity, the tyrosinase inhibitory effect of the extract was 37%, also at $1,000{\mu}g/ml$ concentration. To explore the extractefftoxicity to B16F10 melanoma cells, a 3-[4,5-dimethyl-thiazol-2-yl]-2,5-diphenyl-tetrazoliumbromide assay was performed. Results showed 90% or more cells remained viable at $100{\mu}g/ml$ concentration. A Western blot of the SS extract was used to measure microphthalmia-associated transcription factor (MITF), tyrosinase-related protein-1 (TRP-1), tyrosinase relate protein-2 (TRP-2), and the tyrosinase protein expression inhibitory effect at 25, 50, $100{\mu}g/ml$ concentrations; ${\beta}-actin$ was used as a positive control. Consequently, the MITF, TRP-1, TRP-2, and the tyrosinase protein expression inhibitory effect were seen to decrease by 34.5%, 45.6%, 58.4%, and 79.6%, respectively, at $100{\mu}g/ml$ concentration. These were also then measured by reverse transcription-polymerase chain reaction at 25, 50, $100{\mu}g/ml$ concentrations with GAPDH as a positive control. As a result, the SS extract was seen to decrease MITF, TRP-1, TRP-2, and the tyrosinase protein expression inhibitory effect by 85.4%, 67.5%, 85.2%, 67.1%, respectively at the $100{\mu}g/ml$ concentration. We therefore confirmed the possibility of Sambucus sieboldiana var. pendula (Nakai) extract as a whitening material.

The Study of anti-inflammatory Mechanism with Cobra Venom on Astrocytes of Rats (뇌(腦) 성상세포(星狀細胞)를 대상으로 한 Cobrotoxin의 염증(炎症) 치료(治療) 기전(機轉) 연구(硏究))

  • Yoo, Jae-ryong;Song, Ho-sueb
    • Journal of Acupuncture Research
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    • v.22 no.3
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    • pp.155-167
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    • 2005
  • Objectives : The purpose of this study was to investigate the anti-inflammatory effect of Cobrotoxin on binding affinity of cobrotoxin with P50, $IKK{\alpa}$ and $IKK{\beta}$, activities of NF-${\kappa}B$, Cell viability of astrocyte, expressions of protein molecules of NF-${\kappa}B$ such as P50, P-$1{kappa}B$, $1{\kappa}B$ and iflammation related genes such as Cox-2, iNOS, cPLA2 in the SNP or LPS induced Inflammatory pathway of Rats' astrocytes. Methods : In this study, The expression of cytosolic phospholipase A2, Nitric oxcide, Cyclooxygenase-2 and inducible nitrogen oxide synthase was determined by western blotting with corresponding antibodies, and the generation of NF-${\kappa}B$ was assayed by EMSA method in astrocytes of rats. The Cell viability of astrocytes was determined by MTT assay, and Binding affinity of Cobrotoxin with P50, $IKK{\alpha}$ and $IKK{\beta}$ was assayed by Surface plasmon resonance analysis, and NF-${\kappa}B$ dependent luciferase activity was determined by luciferase analysis, and Uptake of cobrotoxin in astrocytes was identified by Confocal laser scanning microscope Results : 1. Compared with control, LPS-induced NF-${\kappa}B$ DNA binding activity was decreased significantly by 0.1, $0.5{\mu}g/m{\ell}$ of Cobrotoxin in Astrocyte. 2. Compared with control, LPS-induced NF-kB dependent luciferase expression was decreased significantly by 0.1, 0.5 and $1{\mu}g/m{\ell}$ of Cobrotoxin in Astrocyte. 3. Compared with control, SNP induced P50, $I{\kappa}B$ expressions in astrocyte were decreased significantly by 0.1, 0.5 and $1{\mu}g/m{\ell}$ of Cobrotoxin and P-$1{\kappa}B$ expression was decreased significantly by 0.5 and $1{\mu}g/m{\ell}$ of Cobrotoxin. 4. Compared with control, LPS induced P50, $1{\kappa}B$ expressions in astrocyte were decreased significantly by 0.5 and $1{\mu}g/m{\ell}$ of Cobrotoxin. 5. Compared with control, SNP induced Cox-2, iNOS, CPLA2 expressions in astrocyte were decreased significantly by $1{\mu}g/m{\ell}$ of Cobrotoxin. 6. Compared with control, LPS induced Cox-2, cPLA2 expressions in astrocyte were decreased significantly by 0.1, 0.5, $1{\mu}g/m{\ell}$ of Cobrotoxin and iNOS expression was decreased significantly by 0.5, $1{\mu}g/m{\ell}$ of Cobrotoxin. 7. Compared with $0.5{\mu}g/m{\ell}$ of Cobrotoxin, SNP-induced NF-${\kappa}B$ DNA bindins activity in astrocyte was increased significantly by Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 1mM and Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 5mM. 8. Compared with $0.5{\mu}g/m{\ell}$ of Cobrotoxin, LPS-induced NF-${\kappa}B$ DNA binding activity in astrocyte was increased significantly by Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 1mM, Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 5mM, Cobrotoxin $0.5{\mu}g/m{\ell}$with GSH 1mM and Cobrotoxin $0.5{\mu}g/m{\ell}$ with GSH 5mM 9. Compared with $0.1{\mu}g/m{\ell}$ of cobrotoxin, SNP induced P50 expressions in astrocyte were increased significantly by Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 1mM, Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 5mM Cobrotoxin $0.5{\mu}g/m{\ell}$ with GSH 1mM and Cobrotoxin $0.5{\mu}g/m{\ell}$ with GSH 5mM. 10. The uptake of the labeled cobrotoxin into the cells was shown under a confocal laser scanning microscope. cobrotoxin was uptaken into the membrane and nucleus of astrocytes. Conclusions : In summary, the present results demonstrate that cobrotoxin directly binds to sulfhydryl group of p50 and IKKS resulting In the reduction of translocation of p50 and IkB release, thereby inhibits activation of NF-${\kappa}B$, and suggest that pico to nanomolar range of cobrotoxin could inhibit the expression of genes in the NF-${\kappa}B$ signal pathway.

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Effect of Geijibokryunghwan and each constituent herb on inhibition of platelet aggregation (계지복령환(桂枝茯笭丸) 및 그 구성약물(構成藥物)의 혈소판응집억제(血小板凝集抑制)에 관(關)한 연구(硏究))

  • Kim, Jong-Goo;Park, Sun-Dong;Park, Won-Hwan
    • The Journal of Dong Guk Oriental Medicine
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    • v.8 no.2
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    • pp.115-129
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    • 2000
  • The cause that the increase of animality fat intakes, under exercise, fatness, adding the stress, advanced age etc., the occurrence rate of the circulation system disease has been increased. And the thrombosis importantly came to the front as the risk factor of these circulation system's disease. Nowadays, the ischemic disease has especially discussed, for example the angina or myocardial infarction, originated in thrombosis that came from the platelet aggregation. In the western medicine, as the cure and prevention, using the aspirin or ticlopidine for platelet aggregation suppressant. But in the , the curing method must be used properly according to the pectoralgia or heartache's kind, state, grade. The platelet do not attache to the normal hemangioendothelial cell. But when it stimulated by endothelium peronia and so on, it attache to the injury endothelium or rise aggregation between the platelet. On this time, it secrete the platelet aggregation inducer as like ADP, thromboxane A2 from the inside of platelet. So it has first defensive function through the aggregation augment that prevent the celerity consumption of blood. But the activation of abnormal platelet occur the platelet grume and thrombogenesis. So it bring up the occlusive angiosis, so to speak, cardiovascular disease, cerebrovascular disease, arterial sclerosis. In oriental medicine, the thrombosis in the category of blood stasis and this blood stasis present the generalise or local blood circulation disturbance that generated by all kinds of pathological fact or blood stream retention accompanying with a series of syndrome. As the syndrome, stabbing pain fixed at certain region, squamous and dry skin, fullness and pain of the chest and hypochondrium, firmness and fullness of the lower abdomen, black stool, dark purple tongue or with ecchymoses and petechiae etc.. has been created. And it becomes the pathopoiesis cause that the convulsion and palpitation, severe palpitatiion, tympanites, the symtom complex with a mass or swelling in the abdomen, insanity, stricken by wind etc.. Moreover, it used the drugs for invigorating blood circulation and eliminating blood stasis or drugs for removing blood stasis for all kinds of syndrome through the blood stasis. And the drugs for activating the blood circulation, such as Salviae Radix, Angelicae Sinensis Radix, Persicae Semen, Achyranthis Radix, Cnidii Rhizoma, Carthami Flos are used for that. And it is used to the herbs of insects that has strong effect about the disintergrating blood stasis such as Hirudo, Scolopendrae Corpus, Buthus, Lumbricus etc.. On this study, It used Geijibokryunghwan(GBH) and the consisting herbs to investigate the influence of platelet aggregation about drugs that used to improvement various symptoms created by the thrombosis in oriental medicine. GBH formula has as formula recorded in the , action of 'eleminating the evil and not impairment of healthy energy' and 'promoting the flow of QI and cold and heat, so used for the expel blood stasis herbs from the ancient. Therefore we investigated the restraint effect of GBH and the consisting herbs about the platelet agregation induced to the ADP, AA or collagen. The conclusion is following. 1. When it added the aggregation inducer after that it added GBH and individual consisting herbs in the PRP, GBH showed the (+) inhibition effect on the platelet aggregation and it showed the (+) inhibition effect in the individual consisting herbs as like Paeoniae Radix and Moutan Cortex Radicis. 2. It showed the (+), (+,++) inhibition effect on the platelet aggregation in Paeoniae Radix Hoelen, Paeoniae Radix Moutan Cortex Radicis, Hoelen Moutan Cortex Radicis etc. 3. In the aggregation inhibition activating on the difference of density, GBH showed strong inhibition effect to the aggregation state induced to collagen, and it showed the inhibition effect in the individual consisting herbs as like Paeoniae Radix and Moutan Cortex Radicis about the aggregation induced by the collagen. 4. It showed the strong inhibition effect about the aggregation induced by the collagen in Paeoniae Radix Hoelen, Paeoniae Radix Moutan Cortex Radicis, Hoelen Moutan Cortex Radicis etc Like this, as confirm GBH and the individual consisting herb's inhibition effect of platelet aggregation, We considerated that GBH and the individual consisting herbs have practical applicational value of clinical trial in the thrombosis caused by platelet aggregation.

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