• 생약학회지
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• 제30권3호
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• pp.231-237
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• 1999

About forty sea weeds were screened for their inhibitory effects against prolyl endopeptidase, tyrosinase and thrombus coagulation. Out of them, methanolic extract of Ecklonia cava, Sargassum patens, Sargassum hemiphyllum, Sargassum thunbergii, Sargassum singgildianum, Hizikia fusiformis, and Ishige okamurae inhibited more than 90% of prolyl endopeptidase activity at 40 ppm. Sargassum siliquastrum and Ecklonia cava exhibited 51% and 76% of inhibitory activity against tyrosinase at 40 ppm, respectively. In APTT assay system, Sargassum singgildianum, Pterocladia capilacea and Hizikia fusiformis delayed coagulation of thrombus about two times (210, 211, and 198% over control at ca 367 ppm, respectively) and in TT assay, Lomentaria catenata, Laurencia okamurae, and Hizikia fusiformis did most effectively (216,197, and 251% at ca 367 ppm, respectively).

• Applied Biological Chemistry
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• 제39권2호
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• pp.159-164
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• 1996

혈액 항응고 활성물질을 검색하기 위하여 73종의 한약재, 41종의 산채류 및 일반 채소류, 5종의 해조류들의 열수추출물들에 대해 혈액응고 지연시간(Tr)을 시험하였다 1차검색에서 비교적 활성이 높았던 택사, 삼칠근, 현호색, 운지, 마늘, 청각의 산성 수추출물에서 가장 높은 항응고 활성(대조군에 비해 4.3배)을 보였다. 청각(1.2KG)을 0.8% 염산 수용액(24l)으로 추출한 후 메탄을 환류, 에탄올 환류, 침전 및 투석하여 활성이 약 2배 증가된 조다당 CF-1을 얻었다. CF-1은 80.8%의 다당과, 8.7%의 단백질 및 13.3%의 황산기로 구성되어 있었으며, 다당류는 arabinose, galactose, 및 glucose가 주구성당으로 확인되었다. 이 조다당 CF-1의 항응고 활성 본체를 확인하기 위해 pronase처리와 periodate 산화를 행한 결과 pronase 처리시에는 활성의 변화가 없었으나 periodate 산화시에는 약 70%로 활성이 감소한다는 사실로부터 CF-1의 항응고 활성은 다당에 의해 기인됨을 알 수 있었다.

• 한국동물위생학회지
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• 제34권2호
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• pp.191-193
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• 2011

A 3-month-old intact male, Labrador retriever was presented with the history of coagulopathy and anemia. The results of initial screening tests of the hemostatic system yielded a tentative diagnosis of hemophilia. Activated partial thromboplastin time (APTT) was distinctly prolonged (106 seconds) and prothrombin time (PT) was not detected due to markedly prolonged test time. Whole blood transfusions (20 me l/kg body weight) were carried out prior to assays of coagulation factor. After transfusion, the patient recovered well and hemorrhage ceased. Blood samples were assessed for coagulation factor activity. The patient showed markedly low factor IX coagulation activity (5%, reference range: 7~140%) and was diagnosed with hemophilia B. After recovery, the patient was discharged from the hospital. However, 4 months later the patient was re-hospitalized for recurrence of the initial symptoms. The owner did not want to pursue further treatment and the patient died of respiratory distress two days later.

• BMB Reports
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• 제29권6호
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• pp.500-506
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• 1996

We have isolated a water-extracted novel regulator for blood coagulation from an earthworm, Lumbricus rubellus. As a folk remedy, the earthworm has been known to facilitate blood circulation. After complete heat inactivation of endogenous proteases in the earthworm, an anticoagulant(s) was purified through ammonium sulfate fractionation and three consecutive gel permeation chromatography of Sephacryl S-300, Sephadex G-75, and G-150 by measuring activated partial thromboplastin time (APTT) The anticoagulant was further purified to 2,800 fold with a C4 reversed-phase HPLC This activity was stable under heat ($100^{\circ}C$ for 30 min) and acidic conditions (0.4 N HCl). The effects of this partially purified anticoagulant on thrombin were observed with various substrates such as N${\alpha}$-benzoyl-DL-arginine-p-nitroanilide (BApNA), H-D-phenylalanyl-L-pipecoyl-L-arginine-p-nitroanilide (S-2238), N${\alpha}$-p-tosyl-L-arginine methyl ester (TAME), and fibrinogen as a natural substrate. Only TAME hydrolysis, due to an esterase activity of the enzyme, was inhibited among the chromogenic substrates. In addition, the anticoagulant not only inhibited the conversion of fibrinogen to fibrin but also prolonged the fibrin clot formation monitored with the in vitro coagulation test. Based on these observations, we suggest the significance of measuring the ability of antithrombotic drugs to inhibit the esterase activity of thrombin. In this report, it was also shown that the earthworm indeed contained a water-extractable, heat- and acid-stable anticoagulant which could be used as a novel antithrombotic agent.

• 약학회지
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• 제43권6호
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• pp.809-817
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• 1999

The effect of 2-(4-cyanophenyl)amino-1,4-naphthalenedione-3-pyridinium perchlorate (PQ5) on pla-telet aggregation and its action mechanisms were investigated with rat platelet. PQ5 inhibited the platelet aggregation induced by collagen ($6{\;}{\mu\textrm{g}}/ml$), thrombin (0.4 U/ml) and A23187 ($3{\mu}M$) in concentration-dependent manner with $IC_{50}$ values of 5.50, 25.89 and $37.12{\;}{\mu}M$, respectively. PQ5 also significantly reduced the thromboxane $A_2$ (TXA2) formation in a concentration dependent manner. The collagen-induced arachidonic acid (AA) release in [-3H]-AA incorporated platelet, an indication of the phospholipase $A_2$ activity, was decreased by PQ5 pretreatment PQ5 significantly inhibited the activity of thormboxane synthase only at high concentration ($100{\mu}M$), but did not affect the cyclooxygenase activity at all. Collagen-induced ATP release was significantly reduced by PQ5. Calcium-induced platelet aggregation experiment suggests that the elevation of intracellular free $Ca^{2+}$ concentration ($[Ca^{2+}]_i$) by collagen stimulation is decreased by the pretreatment of PQ5, which is due to the inhibition of calcium release from intracellular store and influx from outside of the cell. PQ5 did not showed the effect of anticoagulation as prothrombin time (PT) or activated partial thromboplastin time (APTT). Form these results, it is suggested that PQ5 exerts its antiplatelet activity through the inhibition of the intracellular $Ca^{2+}$ mobilization and the decrease of the $TXA_2$ synthesis.

• 한국임상수의학회지
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• 제24권2호
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• pp.160-163
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• 2007

Patients often present with spontaneous bleeding, or a bleeding disorder may be discovered when an otherwise healthy dog develops marked bleeding during or after surgery. In this study, we were aimed to elucidate whether the cesarean section in dogs has influence on the coagulation profiles. And we gained the normal data on a panel of screening laboratory tests which allow accurate characterization of a hemostatic defects in dogs. Of the 20 healthy adult dogs, buccal mucosa bleeding time (BMBT) was $83.0{\pm}10.5$ seconds, platelet count was $24.0{\pm}3.5{\times}10^4/{\mu}l$, activated partial thromboplastin time (APTT) was $8.8{\pm}2.0$ seconds, the concentration of fibrinogen was $288.5{\pm}77.9mg/dl$, and the concentration of fibrin degradation products (D-dimer) was <250.0 ng/ml. Coagulation profiles before and after cesarean section of 13 cesarean sectioned dogs were in the normal range and there were no statistical differences in coagulation profiles between normal dogs and cesarean sectioned dogs (p>0.05). The results suggested that labor and cesarean section in healthy dogs did not alter coagulation profiles.

• 동의생리병리학회지
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• 제20권4호
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• pp.957-965
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• 2006

This study was peformed to evaluate antithrombotic activities and anti-inflammatory effects of Kami-BoyangHwanoh-Tang(KBHT). The major findings were summarized as follows. In experiment of anti-thrombotic effect; KBHT inhibited human platelet aggregation induced by ADP and epinephrine as compared with the control group and inhibited pulmonary embolism induced by collagen and epinephrine (inhibitory rate is 50 %). KBHT increased platelet number significantly and also KBHT shortened PT and APTT significantly as compared with the control group in thrombus model induced by dextran. In experiment of anti-inflammatory effect; KBHT inhibited $IL-1{\beta}$, IL-6, $TNF-{\alpha}$, COX-2 and NOS-II mRNA expression as compared with the control group in a concentration-dependent degree, and inhibited NO production significantly at 50, $100\;{\mu}g/^{ml}$, and also inhibited ROS production in a concentration-dependent degree as compared with the control group in RAW 264.7 cell line. KBHT inhibited $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$ production significantly in serum of acute inflammation-induced mice, and decreased $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$ production in spleen tissue, and also decreased IL-6 and $TNF-{\alpha}$ production in liver tissue, but increased $IL-1{\beta}$ production in liver tissue of acute inflammation-induced mice. KBHT increased survival rate at 3rd day in mice with lethal endotoxemia induced by LPS. These results suggest that KBHT can be useful in treating diverse female diseases caused by thrombosis and inflammation such as endometrosis, myoma, pelvic congestion, chronic cervicitis, chronic pelvic inflammatory disease and so on.

• Biomolecules & Therapeutics
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• 제2권4호
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• pp.336-342
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• 1994

The general pharmacological properties of EPO were investigated in various animals administering intravenously and in vitro system. The results were as follows. 1. Central nervous system: EPO at doses of 70, 700, 7000 U/kg showed no effect In mice on general behavior, on strychnine- and pentetrazol-induced convulsion and on acetic acid-induced writhing syndrome. The hexobarbital-induced sleeping time in mice was slightly reduced by EPO at a dose of 7000 U/kg but did not change at doses of 70, 700 U/kg. The body temperature in rats was slightly decreased by EPO at doses of 700, 7,000 U/kg but the change was in normal physiological range. 2. Respiratory and cardiovascular system: EPO showed no effect on respiratory rate, blood pressure, heart rate, femoral blood flow, and electrocardiogram in anesthetized dogs at doses of 70, 700, 7000 U/kg. 3. Smooth muscle: EPO at concentrations of 70, 700 U/ml had no effect on the contractile response of isolated guinea pig ileum to histamine and acetylcholine. 4. Water and electrolytes excretion: EPO at dose above 700 U/kg increased urine volume in rats but did not affect the concentrations of $Na^{+},\;K^{+},\;Cl^{-}$ in urine. 5. Gastrointestinal system: EPO(70, 700, 7000 U/kg) had no effect on the intestinal charcoal meal propulsion 6. Blood coagulation system: The administration of EPO(70, 700, 7000 U/kg) had no effect on the plasma prothrombin time(PT) and activated partial thromboplastin time(APTT) in mice. Platelet aggregation induced by ADP and collagen was not influenced by EPO(70 U/ml, 700 U/ml). The overall results obtained indicated that EPO exerts almost no serious pharmacological effect even at a 100-fold clinical dose(7000 U/kg).

• 대한한방부인과학회지
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• 제20권3호
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• pp.45-64
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• 2007

Purpose: This study was performed to evaluate anti-thrombotic and anti-inflammatory effects of NeungaSoJeokTang water extract (NSJT). Methods: In the study of anti-inflammatory effects, NSJT was investigated using cultured cells and murine models. As for the parameters of inflammation, levels of several inflammatory cytokines and chemical mediators which are known to be related to inflammation were determined in mouse lung fibroblast cells(mLFC) and RAW 264.7 cells. Results: Prior to the experiment, we evaluated sGOT, sGPT, BUN and creatine after the treatment. As a result, NSJT was innoxious on liver and kidney. In experiment of anti-thrombotic effect, NSJT inhibited the platelet aggregation induced by ADP and epinephrine, and inhibited pulmonary embolism induced by collagen and epinephrine. NSJT did not affect significantly the blood flow rate both in vitro and in vivo. NSJT increased platelet number and fibrinogen amount, and NSJT shortened PT and APTT in thrombus model induced by dextran. In experiment of anti-inflammatory effect, NSJT inhibited $IL-1{\beta}$, IL-6, $TNF-{\alpha}$, COX-2 and NOS-II mRNA expression in a concentration-dependent manner in RAW 264.7 cell line, and inhibited significantly NO production at 50, 100 ${\mu}g/ml$, and also inhibited ROS production in a concentration-dependent manner. NSJT inhibited $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$ production significantly in serum of acute inflammation-induced Balb/c mice, and decreased $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$ production in spleen tissue, but increased $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$ production in liver tissue. NSJT increased survival rate at the 3th day in ICR mice with lethal endotoxemia induced by LPS. Conclusion: These results suggest that NSJT can be used for treating diverse female diseases caused by thrombosis and inflammation such as pelvic pain, pelvic inflammatory disease as well as vulvar pain due to vulvitis, vulvar vestibulitis and so on.

• 대한의생명과학회지
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• 제8권3호
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• pp.115-125
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• 2002

Gram-negative septicemia, which continues to be a serious clinical problem, is one of the major causes of morbidity and mortality in hospitalized patients. Endotoxin of gram-negative bacteria is a pivotal pathogen of sepsis. To understand the effect of endotoxin on hematological aspect and the time course in early childhood, this study was designed with experimental septic model of young rabbits (8 week-old). Rabbits were divided into control (n=7) and endotoxin group (0.50 mg/kg of endotoxin). The endotoxin group was subdivided into six groups by the sampling times: 3, 6, 12, 24, 48 and 72 hr-group (E-G$_{3}$, E-G$_{6}$, E-G$_{12}$, E-G$_{24}$, E-G$_{48}$ and E-G$_{72hrs}$, each n=7). The evaluation of CBC, activated partial thromboplastin time (APTT), prothrombin time (PT), fibrinogen concentration, coagulation factors and D-dimer were taken from the bloods. The number of leukocytes was lower in E-G$_{3}$ and E-G$_{6hrs}$ (due to pantocytopenia), whereas it was higher in E-G$_{24}$ and E-G$_{48}$ (due to neutrophilia and/or lymphophilia) than in control group (P<0.05). Platelet counts in E-G$_{3}$, E-G$_{6}$, E-G$_{12}$, E-G$_{24}$ and E-G$_{48hrs}$ were lower than those of control group (P<0.05). Normoblast counts in E-G$_{3}$, E-G$_{6}$, E-G$_{12}$, E-G$_{24}$ and E-G$_{48hrs}$ were higher than those of control group (p<0.01). APTT in E-G$_{3}$, E-G$_{6}$, E-G$_{12}$, E-G$_{24}$ and E-G$_{72hrs}$ were longer while PT in E-G$_{3}$, E-G$_{6}$, E-G$_{48}$ and E-G$_{72hrs}$ were higher than those of control group (p<0.05). Fibrinogen concentrations were lower in E-G$_{3}$, E-G$_{6}$ and E-G$_{12}$ but higher in E-G$_{48}$ and E-G$_{72hrs}$ than those of control (p<0.05). Intrinsic coagulation factors (XII, XI, IX, VIII) in all endotoxin groups were significantly lower than those of control group (p<0.05). Extrinsic coagulation factor (X, VII, V, II) were lower in E-G$_{3}$, E-G$_{6}$, E-G$_{12}$ and E-G$_{24hrs}$ whereas they were higher in E-G$_{48}$ and E-G$_{72hrs}$ than in control group (p<0.05). D-dimer concentrations in E-G$_{48}$ and E-G$_{72hrs}$ were higher than those of control group (P<0.001). We concluded that endotoxin led to extensive hematological disturbances including disseminated intravascular coagulation in the young rabbits and that this pathologic condition in the infant and childhood groups will cause the grave results.