• Title/Summary/Keyword: Platelet activation

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Effects of cyclopiazonic acid and aflatoxin B1 on arachidonic acid metabolism, calcium mobilization and ultrastructure in rabbit platelet aggregation (Cyclopiazonic acid 및 aflatoxin B1이 토끼의 혈소판에서 arachidonic acid 대사, 칼슘 동원 및 초미세구조에 미치는 영향)

  • Hong, Choong-man;Jang, Dong-deuk;Cho, Myung-haing
    • Korean Journal of Veterinary Research
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    • v.36 no.4
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    • pp.873-886
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    • 1996
  • For better understanding the interrelationship of hemorrhage and aggregation mechanism, cyclopiazonic acid(CPA) known as promoting the aggregation of platelet, aflatoxin $B_1(AFB_1)$ inhibiting platelet aggregation were used as toxic mycotoxins in these studies. In order to investigate the potential role of prostaglandin metabolism on the platelet aggregation, a variety of prostaglandin metabolites such as $PGF_{2{\alpha}}$, $PGE_2$ and $TXB_2$ were measured in homogenized rabbit platelets by TLC and LSC. And the role of $Ca^{{+}{+}}$ on the platelet aggregation was investigated by flow cytometer. Finally, the morphological effects of mycotoxins on platelet were determined by transmission electron microscope. The results and conclusions obtained from these studies are: 1) CPA induced no changes but $AFB_1$ increased $PGE_2$ and $TXB_2$. 2) CPA promoted ADP, collagen, thrombin, A.A., and PAF-induced $Ca^{{+}{+}}$ release. $AFB_1$, however, decreased $Ca^{{+}{+}}$ level except collagen-induced $Ca^{{+}{+}}$ release. When the calcium blocker, verapamil, was used, CPA decreased thrombin-induced $Ca^{{+}{+}}$ release and increased collagen, ADP, PAF and A.A.-induced $Ca^{{+}{+}}$ release. $AFB_1$ in contrast decreased the all factors induced $Ca^{{+}{+}}$ release. 3) $AFB_1$ did not induce any ultrastructural changes except large vacuole formation in a few platelets. And CPA also did not induce any changes except moderate shape change, indicator of platelet activation. In conclusion, CPA promoted platelet aggregation by the increases of $Ca^{{+}{+}}$ release but had no changes in A.A. metabolites. Antiaggregating effects of $AFB_1$ may be due to decreases of $Ca^{{+}{+}}$ release and increases of $PGE_2$ and $PGF_{2{\alpha}}$ formation. These data provide the basis for the future study of mobilization and function of $Ca^{{+}{+}}$ in platelet aggregation.

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20(S)-ginsenoside Rg3 inhibits glycoprotein IIb/IIIa activation in human platelets

  • Kwon, Hyuk-Woo
    • Journal of Applied Biological Chemistry
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    • v.61 no.3
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    • pp.257-265
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    • 2018
  • The Panax ginseng Mayer is used in conventional medicine in Asia owing to its preventing effects on thrombosis, hypertension, atherosclerosis, vasorelaxation and myocardial infarction. Because platelets are crucial mediators of cardiovascular diseases, many studies have investigated its functions. The previous study showed the antiplatelet effects of crude ginseng fraction and two of its components, ginsenoside Rg3 (20S and 20R). In addition, ginsenoside Rg3-enriched fraction shows an inhibitory effect on collagen-activated rat platelets. However, the mechanism underlying this effect remains unclear. Thus, I investigated the inhibitory action of ginsenoside Rg3 (20S, G-Rg3) on the regulation of signaling molecules involved in ${\alpha}IIb/{\beta}_3$ activation. I found that G-Rg3, in a cyclic AMP dependent manner, inhibited thrombin-induced activation of human platelets and affinity of fibrinogen and fibronectin with ${\alpha}IIb/{\beta}_3$. Thus, in the present study, G-Rg3 showed an inhibitory effect on glycoprotein IIb/IIIa (${\alpha}IIb/{\beta}_3$) activation, suggesting its potential use for preventing platelet-mediated thrombotic disease.

Activation of Platelet Rich Plasma by Soluble Canine Small Intestinal Submucosa Gel and Bovine Thrombin (개 소장점막하 겔과 소 트롬빈을 이용한 혈소판풍부혈장의 활성화 연구)

  • Lee, A-Jin;Lee, Changsun;Kim, Hyun;Chung, Dai-Jung;Do, Sun Hee;Kim, Hwi-Yool
    • Journal of Veterinary Clinics
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    • v.31 no.3
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    • pp.170-174
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    • 2014
  • This study evaluated the efficacy of soluble canine small intestinal submucosa gel in comparison to bovine thrombin in activating rabbit platelet rich plasma (PRP) by detecting growth factors. PRP from rabbits was activated by using soluble canine SIS gel, bovine thrombin, or both. The surface morphology of each group of samples was examined by scanning electron microscopy. The release of transforming growth factor (TGF)-${\beta}1$ from each set of samples was measured over 7 days using enzyme-linked immunosorbent assay. The PRP-canine SIS gel group exhibited the highest total amount of released TGF-${\beta}1$. However, there were no significant differences between any groups. The use of soluble type of canine SIS gel could be an effective alternative to bovine thrombin.

Platelet-Activating Factor Enhances Experimental Pulmonary Metastasis of Murine Sarcoma Cells by Up-regulation of Matrix Metalloproteinases-9 Through NF-$\kappa$B-Dependent Pathway

  • Ko, Hyun-Mi;Back, Hae-Kyong
    • Biomedical Science Letters
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    • v.10 no.2
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    • pp.143-151
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    • 2004
  • Matrix metalloproteinases (MMPs) are capable of degrading extracellular matrix, a process that is necessary for angiogenesis, tumor invasion and metastasis. Platelet-activating factor (PAP) increases angiogenesis, tumor growth and metastasis through nuclear factor (NF)-$\kappa$B activation. Based on these facts, the involvement of MMPs in PAF-induced pulmonary metastasis was investigated in murine sarcoma cells, MMSV-BALB/3T3. Messenger RNA expression and enzymatic activity of MMP-9 were assessed by RT-PCR and zymography, and cell migration and metastasis were done for the detection of MMP-9 functional activity. PAP induced mRNA expression and enzymatic activity of MMP-9, and its effects were either inhibited by the PAP antagonist, WEB 2170 or by the NF-$\kappa$B inhibitor, parthenolide, or p65 antisense oligonucleotide in a dose-dependent manner. In addition, PAF induced promoter activity of MMP-9, which was inhibited by WEB 2170, phenanthroline, NAC, PDTC. These results indicate that PAF induces mRNA expression and enzymatic activity of MMP-9 in NF-$\kappa$B dependent manner. Cell migration assay showed that PAF induced MMSV-BALB/3T3 migration, and its effect was significantly inhibited by treatment with phenanthroline. PAF enhanced pulmonary metastasis of murine sarcoma cells, MMSV-BALB/3T3 was also reduced by phenanthroline. These results suggest that PAF-enhanced cell migration and pulmonary metastasis is mediated through the expression of MMP. In conclusion, It is suggested that PAF enhances pulmonary metastasis by inducing MMP-9 expression via the activation of NF-$\kappa$B.

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Effects of Shikonin Pigments from the Roots of Lithospermum eryhrorhizon on Rabbit Platelets (자근으로부터 혈소판에 작용하는 천연색소물질에 관한 연구)

  • 박영현;장성근
    • Journal of Food Hygiene and Safety
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    • v.15 no.2
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    • pp.167-172
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    • 2000
  • Lithospermum erythrorhyzon has been used as a red fooddye and traditional Chinese medicine to treat wounds, skin diseases and burns. Platelet activation plays an important role in thrombosis and haemostasis. Here, we studied the inhibition of platelet activation and its active compound from the root of Lithospermum erythrorhyzon. Its ethyl acetate extract inhibited the aggregation of washed rabbit platelets induced by collagen or thrombin. Five naphthoquinone pigments , shikonin , acetylshikonin , is obutylshikonin, $\alpha$-methyl-n-butylshikonin and $\beta$,$\beta$-dimethylacrylshikonin were isolated by means of high pressure liquid chromatography. The structures were determined by comparison of their proton nuclear magnetic resonance spectra. The potency of their inhibition was in the following order : $\beta$,$\beta$-dimethylacrylshikonin$\geq$$\alpha$-methyl-n-butylshikonin>isobutylshikonin>acetylshikonin>shikonin. It is suggested that the size of the aliphatic hydroxy group of shikonin is important for the enhancement of potency.

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Effect of Extract from Paeoniae radix on Rabbit Platelets (작약 추출물의 혈소판 작용에 관한 연구)

  • 박영현;손동주
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.31 no.3
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    • pp.511-515
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    • 2002
  • Paeoniae radix has been considered as one of the most important crude drugs used in traditional oriental medicine and has been employed as a circulatory tonic in care of weakness, night sweats, and lumbar pain, etc. Platelet activation plays an important role in thrombosis and haemostasis. Active compounds for the inhibition of platelet activation from Paeoniae radix were extracted and fractionated into five fractions. Its fraction two and three of ethyl acetate extract inhibited the aggregation of washed rabbit platelets induced by collagen. Two active compounds, bezoyloxypaeoniflorin and paeoniflorin, were isolated from fraction two and three by silica gel column and high performance liquid chromatography. The chemical structures were determined by comparison of their proton and carbon nuclear magnetic resonance spectra. Benzoyloxypaeoniflorin showed strong inhibition at the concentration of 100 ug/mL against collagen-induced washed rabbit platelets aggregation. It is suggested that Paeoniae radix has become food material to prevent a cardiovascular disease.

Study on Inhibition of Platelet Aggregation of Bioactive Constituents from Paeonia lactiflora (작약의 혈소판 응집억제작용에 관한 연구)

  • 박관혁;서범석;손동주;박영현;장성근
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.4 no.4
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    • pp.357-360
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    • 2003
  • Methanol extracts from Paeonia lactiflora showed a strong inhibition against platelet aggregation on platelet activation test. Therefore, the bioactive constituents from Paeonia lactiflora were prepared using chromatography methods and were analyzed by NMR and reference data. Compound 1b was confirmed a same structure with henzoyloxypaeoniflorin, compound 2e was a same structure with paeoniflorin; main product of Paeonia lactiflora. Analytical data of compound 3a were not consistent with any known paeoniflorin soucture, but showed the souctural similarity with it. And also the aggregation inhibition activity of compound 3a showed a strong inhibition($\geq$ 90%) induced by collagen. Therefore it suggested that the structure of compound 3a may be the similar structure of benzoyloxypaeoflorin with a functional group in place of benzoyl group and/or a different functional group in stead of Rl. We suggested that benzoyl group of benzoyloxypaeoniflorin substitued instead of 5-carbon OH group on glycoside moiety paeoniflorin played role of the metabolite in case of a platelet aggregation inhibition activity. Paeoniflorin showed more strong inhibition by thrombin than collagen. Therefore, it may be destructed a calcium metabolite as a forming $Ca^2+$ chelate. Compound 3a may be that other functional group instead of OH group of 5-carbon on glycoside moiety of paeoniflorin and/or OH group of benzoyl moiety of paeoniflorin played role of the metabolite in a platelet aggregation inhibition.

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Decreased Expression of Phospholipase C-$\beta$2 in Human Platelets with Impaired Function

  • Lee, Sang-Bong;A. Koneti Rao;Lee, Kweon-Haeng;Xu Yang;Bae, Yun-Soo
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.11a
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    • pp.75-84
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    • 1996
  • Platelets from a patient with a mild inherited bleeding disorder and abnormal platelet aggregation and secretion show reduced generation of inositol 1,4,5-trisphosphate (IP$_3$), mobilization of intracellular Ca$\^$2+/, and phosphorylation of pleckstrin in response to several G protein mediated agonists, suggesting a possible defect at the level of phospholipase C (PLC) activation. A procedure was developed that allows quantitation of platelet PLC isozymes. After fractionation of platelet extracts by high-performance liquid chromatography, seven, out often known PLC isoforms were detected by immunoblot analysis. The amount of these isoforms in normal platelets decreased in the order PLC-${\gamma}$2 > PLC-${\beta}$2 > PLC-${\beta}$3 > PLC-${\beta}$l > PLC-${\gamma}$ > PLC-$\delta$1 > PLC-${\beta}$4. Compared with normal platelets, platelets from the patient contained approximately one-third the amount of PLC-${\beta}$2, whereas PLC-${\beta}$4 was increased threefold. These results suggest that the impaired platelet function in the patient in response to multiple G protein mediated agonists is attributable to a deficiency of PLC-${\beta}$2. They document for the first time a specific PLC isozyme deficiency in human platelets and provide an unique opportunity to understand the role of different PLC isozymes in normal platelet function.

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