• Title/Summary/Keyword: Embryo Extracts

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Saururus chinenesis Extracts Scavenge Reactive Oxygen Species and Modulate Nitric Oxide Production in Raw 264.7 Macrophages

  • Oh, Jang-Hee;Shon, Hee-Kyoung;Oh, Moon-You;Chung, An-Sik
    • Toxicological Research
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    • v.18 no.2
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    • pp.117-127
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    • 2002
  • Saururus chinensis Baill has been used in Korean folk medicine for the treatment of various diseases such as edema, Jaundice, and furuncle. The components of this plant were extracted into four fraction. Among the four fraction, hexane and ethyl acetate fraction were highly toxic to 3T3 mouse embryo fibroblast and Raw 264.7 mouse macrophage, but n-butanol and residue fraction did not show any toxic effect to those cell lines. n-Butanol and residue fraction exhibited antioxidant effects on hydro-gen peroxide, hydroxyl radical, and superoxide anion directly in vitro and in the 3T3 fibroblasts. All the four fractions inhibited lipid peroxidation measured by thiobarbituric acid reactive substances (TBARS) formation. In addition, n-butanol and residue fraction showed inhibitory effects on lipopolysaccharide (LPS)-induced nitric oxide production, and also down-regulated inducible nitric oxide synthase (iNOS) mRNA transcription 6 h after LPS stimulation in Raw 204.7 cells. Only n-butanol fraction, which mainly consists of flavonoids, inhibited NF-kB activation by decreasing IkBa degradation 90 min after LPS stimulation. horn the results, it is suggested that this plant could be a good candidate material for drug development based on its antioxidant and/or anti-inflammatory constituents.

Cloning of a Tissue Inhibitor of Matrix Metalloproteinase-1 (TIMP-1) from a Scylliorhinus torazame (두툽상어에서 tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) 유전자의 클로닝)

  • Kim, Cha-Soon;Bae, Su-Kyoung;Kim, Kyu-Won;Kim, Yung-Jin
    • Journal of Life Science
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    • v.6 no.4
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    • pp.286-292
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    • 1996
  • Angiogenesis is a fundamental process by which new blood vessels are formed. It is essential in embryo development, and wound healing. Furthermore, malignant tumor growth and metastasis are also angiogenesis-dependent. In the catilage tissue, normal angiogenesis process is suppressed. In fact, it was reported that angiogenesis-inhibitory substances were isolated from the extracts of cow and shark catilage tissue. In order to isolate genes involved in the regulation of angiogenesis from a catilage fish, we constructed a shark cDNA library from Scylliohinus torazame. We then screened the library using hyman tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) gene as a probe. Among the 4 X 10$^{4}$ plaques screened, we isolated 2 positive clones (T-1, T-2). Restriction enzyme analysis revealed that the T-1 clone contains 0.8 kb cDNA insert, and the T-2 clone contains 1.2 kb and 2.2 kb inserts, respectively. Further DNA sequence analysis shows that the DNA sequence of the T-1 clone is 53% homologous to that of the human TIMP-1 gene.

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Methylation Changes of Lysine 9 of Histone H3 during Preimplantation Mouse Development

  • Yeo, Seungeun;Lee, Kyung-Kwang;Han, Yong-Mahn;Kang, Yong-Kook
    • Molecules and Cells
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    • v.20 no.3
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    • pp.423-428
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    • 2005
  • Immediately after fertilization, a chromatin remodeling process in the oocyte cytoplasm extracts protamine molecules from the sperm-derived DNA and loads histones onto it. We examined how the histone H3-lysine 9 methylation system is established on the remodeled sperm chromatin in mice. We found that the paternal pronucleus was not stained for dimethylated H3-K9 (H3-$m_2K9$) during pronucleus development, while the maternal genome stained intensively. Such H3-$m_2K9$ asymmetry between the parental pronuclei was independent of $HP1{\beta}$ localization and, much like DNA methylation, was preserved to the two-cell stage when the nucleus appeared to be compartmentalized for H3-$m_2K9$. A conspicuous increase in H3-$m_2K9$ level was observed at the four-cell stage, and then the level was maintained without a visible change up to the blastocyst stage. The behavior of H3-$m_2K9$ was very similar, but not identical, to that of 5-methylcytosine during preimplantation development, suggesting that there is some connection between methylation of histone and of DNA in early mouse development.

Studies on the Effect of the Protein Constituents of Panax ginseng Root on Cultured Chick Embryonic Brain, Spinal Cord and Skeletal Muscle Cells (인삼 단백성분이 배양한 Chick Embryo의 뇌, 척수, 근육세포에 미치는 효과에 관한 연구)

  • Kim, Young-Choong;Han, Dae-Suk;Huh, Hoon;Ahn, Sang-Mee;Koo, Hyang-Ja
    • YAKHAK HOEJI
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    • v.27 no.2
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    • pp.109-116
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    • 1983
  • The effect of protein constituents of six-year old fresh Panax ginseng root on chick embryonic brain, spinal cord and skeletal muscle dissociation cultures was studied. The protein constituents showed the enhancing effect on cultured brain, spinal cord and skeletal muscle cells. The neurite formation from brain and spinal cord cells and the outgrowth of neurite seemed to be enhanced by almost all of the protein constituents employed for this study. The maturation of skeletal muscle cells was stimulated by the protein constituents. This enhancing effect of the protein constituents was more vivid when brain, spinal cord and skeletal muscle cells were cultured with a medium which did not contain chick embryonic extracts known as an essential component for primary cell culture. The protein fraction having molecular weight range of 1,000 to 5,000 out of all the protein fractions employed for this study showed the most stimulatory effect on cultured brain, spinal cord and skeletal muscle cells.

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A study on the inflammatory response induced by LPS of the Arthrospira platensis ethanol extract

  • Zhang, Shi Jie;Yang, Jae-Chan;Kim, Bo-Ae
    • Journal of the Korean Applied Science and Technology
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    • v.36 no.3
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    • pp.966-974
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    • 2019
  • Arthrospira platensis has been reported to contain a variety of substances such as phycocyanin, ${\beta}$-carotene, vitamin E and other carotenoids. In this study, zebrafish were treated with indoor cultivation spirulina ethanol extracts(ICAE) to determine toxicity(coagulation rate, hatching rate, heart rate). We used the DCFH-DA staining method to detect the effect of reactive oxygen species(ROS) generation on lipopolysaccharide(LPS)-induced zebrafish embryos ROS various concentrations(0.01, 0.05, 0.1, 0.5mg/ml) of ICAE. Cell toxicity was measured by WST-1 assay on RAW 264.7 macrophage cell line. Also, measured the inhibitory effect of nitric oxide(NO) and prostaglandin $E_2$ ($PGE_2$) production in RAW264.7 macrophages induced by LPS at various concentrations of ICAE. The results of embryo coagulation rate, hatching rate, heart rate of zebrafish at various(0.01, 0.05, 0.1mg/ml) of ICAE was no toxicity. The ICAE treated group had an inhibitory effect on NO and $PGE_2$ production compared and decreased with concentration. The results of this study ethanol extract of Arthrospira platensis has an anti-inflammatory effect and suggest that is worthy of use cosmetics for skin protection.

Anti-Angiogenic Activity of Gecko Aqueous Extracts and its Macromolecular Components in CAM and HUVE-12 Cells

  • Tang, Zhen;Huang, Shu-Qiong;Liu, Jian-Ting;Jiang, Gui-Xiang;Wang, Chun-Mei
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.5
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    • pp.2081-2086
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    • 2015
  • Gecko is a kind of traditional Chinese medicine with remarkable antineoplastic activity. However, undefined mechanisms and ambiguity regarding active ingredients limit new drug development from gecko. This study was conducted to assess anti-angiogenic properties of the aqueous extracts of fresh gecko (AG) or macromolecular components separated from AG (M-AG). An enzyme-linked immunosorbent assay (ELISA) approach was applied to detect the vascular endothelial growth factor (VEGF) secretion of the tumor cells treated with AG or M-AG. The effect of AG or M-AG on vascular endothelial cell proliferation and migratory ability was analyzed by tetrazolium dye colorimetric method, transwell and wound-healing assays. Chick embryo chorioallantoic membrane (CAM) assays were used to ensure the anti-angiogenic activity of M-AG in vivo. The results showed that AG or M-AG inhibited the VEGF secretion of tumor cells, the relative inhibition rates of AG and M-AG being 27.2% and 53.2% respectively at a concentration of $20{\mu}L/mL$. AG and M-AG inhibited the vascular endothelial (VE) cell proliferation with IC50 values of $11.5{\pm}0.5{\mu}L/mL$ and $12.9{\pm}0.4{\mu}L/mL$ respectively. The VE cell migration potential was inhibited significantly (p<0.01) by the AG (${\geq}24{\mu}L/mL$) or M-AG (${\geq}12\mu}L/mL$) treatment. In vivo, neovascularization of CAM treated with M-AG was inhibited significantly (p<0.05) at a concentration of ${\geq}0.4{\mu}L/mL$. This study provided evidence that anti-angiogenesis is one of the anti-tumor mechanisms of AG and M-AG, with the latter as a promising active component.

Cytotoxicity of Hyaluronic Acid Membrane Cross-linked with Lactide (락타이드로 가교시킨 히아루론산 막의 세포독성)

  • Kim, Won-Jung;Kwon, Ji-Young;Cheong, Seong-Ihl;Kim, In-Seop
    • KSBB Journal
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    • v.21 no.4
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    • pp.255-259
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    • 2006
  • The biodegradable hyaluronic acid(HA) membranes cross-linked with lactide using the crosslinking agent, 1-ethyl-3(3-dimethyl aminopropyl) carbodiimide(EDC) were prepared as a potential biocompatible material for tissue engineering. HA membranes having different mechanical properties were synthesised by varying degree of the mole ratio of lactide to HA, EDC concentration, and crosslinking temperature. HA membranes were degradable in water solution and the degradation became slower with the increasing mole ratio of lactide to HA. HA membranes were sterilized using ethylene oxide gas and extracted with cell culture medium for 24 h at $37^{\circ}C$ and 200 rpm. Cytotoxicity of the extract was tested using NIH/3T3 mouse embryo fibroblast as a model cell. Growth inhibition was not observed in the extracts of HA membranes with the mole ratios of lactide to HA, 5 or 10, and 10% EDC concentration, however 11% of growth inhibition was observed in the extract with the mole ratio of 13. Growth inhibition was not observed in the extracts of HA membranes prepared with 5% EDC or 10% EDC and the mole ratio of lactide to HA, 10, however 12% of growth inhibition was observed in the extract with 20% EDC. Cytotoxicity was not observed in the extracts of HA membranes prepared at varying crosslinking temperatures, $15^{\circ}C,\;25^{\circ}C,\;and\;28^{\circ}C$ with the mole ratio of lactide to HA, 10 and 10% EDC.

A Study on the Antioxidative Effect of Orostachys Japonicus A. Berger Ethyl Acetate Fraction (와송 에틸아세테이트 분획물의 항산화 효능에 관한 연구)

  • Im, Eun Kyung;Yang, Jae Chan
    • Journal of the Korean Applied Science and Technology
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    • v.38 no.1
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    • pp.118-125
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    • 2021
  • In this study Orostachys japonica A. Berger used is a medicinal herb that has long been used as a folk remedy for cancer treatment. In this study, the antioxidant efficacy of the ethyl acetate fraction of Orostachys japonica A. Berger was confirmed. The results of the Orostachys japonica A. Berge ethyl acetate fraction of antioxidant activity assays showed Antioxidant effect of Orostachys japonica A. Berger EtOAc fraction extract at 0.10 mg/mL was showed a DPPH radical scavenging rate of 78.54% and ABTS+ radical scavenging rate of 73.48%. Also, the toxicity result of Orostachys japonica A. Berger EtOAc fraction extracts using alternative experimental animal model zebrafish, confirmed a 100% the survival of the zebrafish embryo was shown that there was no coagulation and no hatching delay at all concentrations. also ROS generation induced by UV-B irradiation was confirmed that the fluorescence intensity decreased as a whole in all larvae treated with Orostachys japonica A. Berger EtOAc fraction extracts. In particular, it was confirmed that ROS generation was effectively suppressed by showing a 35.7% reduction rate compared to the positive control at a concentration of 3 ㎍/mL. These results were confirmed that Orostachys japonica A. Berger EtOAc fraction extracts has the possibility of application in the cosmetics field as a natural antioxidant.

Anti-thrombotic Activities of Hot-water Extracts Prepared from Various Parts of Nelumbo nucifera Gaertner (다양한 부위의 연(Lotus) 열수 추출물의 항혈전 활성)

  • Ahn, Seon-Mi;Sung, Hwa-Jung;Kim, Jong-Sik;Park, Jong-Yi;Sohn, Ho-Yong
    • Journal of Life Science
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    • v.28 no.10
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    • pp.1156-1162
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    • 2018
  • To investigate the anti-thrombotic activities of the lotus (Nelumbo nucifera Gaertner), various hot-water extracts were prepared from the leaf, pod of seed (PS), seed, embryo of seed (ES), root, and the node of root (NR) of the lotus. The highest extraction ratio was found in the NR (20.3%), followed by the seed, root, leaf, ES, and PS. These extracts had pH and acidity levels ranging from 5.6~6.5 and 0.06~0.20%, respectively. The seed extract showed 70% brix, whereas the leaf and PS extracts showed less than 0.1% brix. The highest contents of total polyphenol (179.7 mg/g), total flavonoids (161.4 mg/g), and reducing sugar (161.4 mg/g) were observed in the leaf extract, and the highest total sugar content (873.0 mg/g) in the seed extract. Anti-coagulation assay of the extracts of NR, leaf, and PS showed strong activities. In particular, at a concentration of 5 mg/ml, the PS extract had 15-fold extended thrombin, prothrombin, and activated partial thromboplastin times. However, only the ES extract showed activities inhibitory to platelet aggregation, with treatment with 0.25 mg/ml of ES extract decreasing platelet aggregation to 25.1%, a reduction comparable to that effected by aspirin. The extracts other than the seed extract showed no hemolysis activities against human RBC at treatments of up to 1 mg/ml. These results suggest that the NR, PS, seed, and ES, all byproducts of the lotus agriculture industry, have high potential as novel sources of anti-thrombotic agent.

Effect of Culture Media on Embryonic Cell Growth in Zebrafish, Danio rerio (배지에 따른 제브라피쉬(Danio rerio) 배아 유래세포의 성장 효과에 관한 연구)

  • Lee, Ki-Young;Kim, Jong-Yeon;Jo, Soo-Gun
    • Development and Reproduction
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    • v.12 no.1
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    • pp.51-56
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    • 2008
  • To optimize the cell culture conditions of zebrafish embryonic cells, we compared the efficiency of three types of medium, DMEM, K-NAC and D-NAC. In this study, we showed that the cells grown in K-NAC have better plating efficiency than DMEM, especially in the case of low cell seeding density. However, cells grew slower in K-NAC than those in DMEM in confluent cultures. The effect of 0.1% zebrafish embryo extracts was minimal. The presence of 1% trout serum in culture medium significantly increased the growth rate of cells(p<0.05). No difference was found at $2{\sim}3{\times}10^5$ cell seeding density(p<0.05). At $4{\sim}5{\times}10^5$ cell seeding density, cells grew better in DMEM than K-NAC (p<0.1). The results suggest that supplementation of NAC and A2P in Keratinocyte SFM may improves plating efficiency when cells are plated at low population. No difference was found for cell growth in either medium with 5%, 10% or 15% FBS supplemented (p<0.05). Cells culture in D-NAC grew significantly better than those in DMEM(p<0.05). Our results clearly showed that the use of NAC and A2P in the culture medium has a positive effect on cell growth regardless of the amount of FBS added.

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