• The Journal of Internal Korean Medicine
• /
• v.17 no.1
• /
• pp.34-50
• /
• 1996

The present experiment was desinged to investigate the effects of Tongdosan water extracts on the Cardiovascular System in the Experimental Animals. Thus, the changes of blood pressure and heart rate were measured after oral admini- stration. Measurment of Mortality rate was observed for measuring the effect of Tongdosan water extract. Tongdosan water extract against pulmonary thrombo- embolism induced by collagen the mixture(0.1ml/10g, 2mg/kg B.W) plus serotonin(5mg/kg B.W) in mouse. The effect of Tongdosan water extract was examined by observing the change of collagen-induced platelet aggregation, coagulation activity, ex vivo and in vitro fibrinolytic activity of euglobulin fraction in rats. The results were summarized as followings. 1. Tongdosan dropped the blood pressure in spontaneous hypertensive rat. 2. The drug increased the auricular blood flow in rabbit. 3. The drug relaxed the artery contraction by pretreated norepinephrine in rat. 4. The drug inhibited the death rate of mouse which was led to thromboembo- lism by serotonin and collagen. 5. The drug inhibited the platelet aggregation in rat. 6. The drug prolonged the prothrombin time and activated partial thromboplastin time on the test of plasma coagulation factor activity in rat, but was not valuable. 7. The drug reduced the fibrinogen lyses time of rat ex vivo assay and lyses area was increased. 8. Tongdosan reduced fibrinogen lyses time of rat in vitro assay. According to the above mentioned results, Tongdosan increased the blood flow and dropped the blood pressure by the dilation of blood vessel. And the drug presented the antithrombin acivity, inhibited the platelet aggregation.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.18 no.5
• /
• pp.1494-1504
• /
• 2004

The present paper reports the effects of Hwaotang an atherosclerosis using a spontaneous experimental model, We have also investigated the pharmacological effect of Hwaotang on collagen- and ADP-induced blood platelet aggregation, thrombin-induced conversion of fibrinogen and fibrinolysis in in vitro experiments, and various effects on stimuli-induced superoxide generation in human neutrophils. Hwao-tang was shown to have inhibitory effect on collagen- and ADP-induced blood platelet aggregation, on thrombin-induced conversion of fibrinogen to fibrin and on the activity of plasminogen or plasmin. Hwao-tang also significantly inhibited fMLP-induced superoxide generation in a concentration-dependent manner, but not that induced by arachidonic acid. Hwao-tang inhibited neutrophil functions, including degranulation, superoxide generation, and leukotriene B4 production, without any effect on 5-lipoxygenase activity. In conclusion, the protection of extracts of Hwao-tang on the ischemic infarction induced artificially might be involved to their inhibition of thrombotic action. The results also indicate that Hwao-tang exerts the effects on superoxide generation related to the inhibition of neutrophil functions.

• Journal of Society of Preventive Korean Medicine
• /
• v.2 no.1
• /
• pp.193-208
• /
• 1998

In order to study the effects of emitted-qi therapy, Youngyeonondamtang and Jeokyeonondamtang on the mice bearing ascites cancer caused by Sarcoma 180, the author divided the mice into normal group, control group, EQ group, YO group, JO group, CTX+QE group, CTX+YO group, CTX+JO group, and treated with emitted -qi therapy and administrated Youngyeonondamtang, Jeokyeononondaintang and Cyclophosphamide (CTX). After collecting blood from the mice, measured the values of Prothrombin Time(PT), Partial Thromboplastin Time(PTT), Fibrinogen, White Blood Cell(WBC) and Platelet. The results were as follows, 1. PT was not showed any significant change in ever)r group, being compared with the control group. 2. H increased significantly in YO group, being compared with the control group. 3. Fibrinogen decreased significantly in EQ group, YO group, CTX+EQ group, CTX+YO group, CTX+JO group, being compared with the control group. 4. WBC increased significantly in CTX+EQ group, CTX+YO group, CTX+JO group, being compared with the CTX group. 5. Platelet decreased significantly in every group, being compared with the control group. These results suggest that emitted-qi therapy, Youngyeonondamtang and Jeokyeonondamtang had the effects that could heal the hematopoiesis system in the mice bearing ascites cancer, and especially had the better effects in the case that hematopoiesis system has been impaired by CTX.

• The Korean Journal of Mycology
• /
• v.32 no.2
• /
• pp.119-124
• /
• 2004

Metalloenzyme was purified from the fruiting bodies of Tricholoma sejunctum. MALDI-TOF and ICP/MS analyses revealed that the enzyme had a molecular weight of 18788.25 and includes $Zn^{2+}$ ion. The N-terminal amino acid sequence of the enzyme was Ala-Thr-Tyr-Lys-Ile-X-Ser-Ala-Thr-His-Gln-X-X-Leu-Val. The activity of the enzyme was inhibited by EDTA and 1,10-phenanthroline, indicating that the enzyme was a metalloprotease. No inhibition was found with E-64 and pepstatin. It has broad substrate specificity for synthetic peptides. The enzyme was stable up to $40^{\circ}C$. The activity of the enzyme was increased by $Zn^{2+}$ and $Co^{2+}$, while it was totally inhibited by $Hg^{2+}$. The enzyme hydrolyzes $A{\alpha}$ subunit of human fibrinogen but did not show any reactivity for $B{\beta}$ and ${\gamma}$ form of human fibrinogen.

• The Journal of Korean Medicine
• /
• v.16 no.1 s.29
• /
• pp.351-369
• /
• 1995

In order to investigate the effect of Qingbaozhuyutang(淸胞逐瘀湯), Bee Veenom on 'Blood Stasis Pattern', here We regard Thrombosis and Elevated Blood viscosity as Blood Stasis Pattern. rats were injected Endotoxin into the caudal vein to make experimental thrombosis model(control group), and at sample group, rats were orally administrated solid extract of Qingbaozhuyutang(淸胞逐瘀湯) 0.5g/200g or were injected Apitoxin 1mg/Kg into the abdominal region(sample group), and then we observed platelet, fibrinogen, prothrombin time and FDP level. Another group were injected HA into the muscular rump to make experimental elevated blood viscosity instead of Endotoxin. Thereafter we measured whole blood viscosity and plasma viscosity. The results were summarized as follows. 1. Platelets were increased significantly in sample I compared with the control group. 2. Fibrinogen levels were increased significantly in sample I compared with the control group. 3. Prothrombin times were shortened significantly in sample II and sample III compared with the control group. 4. FDP levels were decreased significantly in sample II and sample III compared with the control group. 5. Whole blood viscosity and plasma viscosity were decreased significantly in all shear rates compared with the control group. According to the above results, it is considered that Qingbaozhuyutang(淸胞逐瘀湯) has significant effects on platelets, fibrinogens, prothrombin times, FDP levels, whole blood viscosity and plasma viscosities, therefore it seems to be applicable to disease related to 'Blood Stasis Pattern' through the thrombosis and elevated blood viscosity. But, Veenom has a little effect. the mechanism concerned for the effects is to be investigated in future.

• The Journal of Internal Korean Medicine
• /
• v.18 no.1
• /
• pp.97-113
• /
• 1997

The present experiment was designed to investigate the effects of Sambutang water extracts on the serum cholesterol levels and the cardiovascular system in the experimental animals. Thus, the changes of blood pressure and heart rate were measured after oral administration. Measurment of Mortality rate was observed for measuring the effect of Sambutang water extract. Sambutang water extract against pulmonary thromboembolism induced by collagen the mixture(0.1 ml/10 g, 2 mg/kg) plus serotonin(5 mg/kg) in mouse. The effect of Sambutang water extract was examined by observing the change of collagen-induced platelet aggregation, coagulation activity, ex vivo and in vitro fibrinolytic activity of euglobulin fraction in rats. The results were summarized as follows. 1. Sambutang decreased the serum cholesterol levels in rats. 2. Sambutang dropped the blood pressure in spontaneous hypertensive rat. 3. The drug increased the auricular blood flow in rabbit. 4. The drug relaxed the artery contraction by pretreated norepinephrine in rat. 5. The drug inhibited the death rate of mouse which was led to thromboembolism by serotonin and collagen. 6. The drug inhibited the platelet aggregation in rat. 7. The drug prolonged the prothrombin time and activated partial thromboplastin time on the test of plasma coagulation factor activity in rat, but was not valuable. 8. The drug increased the antithrombin activity in rat and the fibrinogen lysis time was reduced and lysis area was increased. 9. Sambutang reduced fibrinogen lysis time of rat in vitro assay. According to the above mentioned results. Sambutang increased the blood flow and dropped the blood pressure by the dilation of blood vessel. And the drug presented the antithrombin activity, inhibited the platelet aggregation.

• Herbal Formula Science
• /
• v.6 no.1
• /
• pp.199-214
• /
• 1998

This study was performed to prove the antithrombotic effects of DoHongYeum fluid by way of experimental methods. The thrombosis was induced by injection of collagen the mixture (0.1ml/10g, 2mg/kg B.W)plus serotonin (5mg/kg B.W) into the caudal vein of rat, 2 hours after liquid extract of DoHongYeum was oral administration. The effect of the fluid of DoHongYeum was rxamined by the number of RBC and platelets, bleeding time, blood clotting time, death rate, platelet aggregation, plasmacoagulation factor activity, exvivo and fibrinolytic activity of englobulin fracture in the rats. The results were summerized as followings. 1. The number of RBC and platelets was significantly increased in DoHomeYeum group incomparison with the control group. 2. Bleeding time was significantly shortened in DHY group in comparison with the controlgroup. 3. Blood clotting time was significantly prolonged in DHY group in comparison with the control group. 4. The death rate of mouse was inhibited in DHY group in comparison with the control group. 5. The platelet aggregation was inhibited in DHY group in comparison with the control group. 6. The prothrombin time and activated partial thromboplastin time on the test of plasmacoagulation factor activity was prolonged but was not valuable in DHY group. 7. Fibrinogen lyses time of rat was reduced and lyses area was increased in DHY group incomparison with the control group. 8. Fibrinogen lyses time of rat in vitro assay was reduced in DHY group. From the above results, it was thought that the DoHongYeum could be applied effectively in the thrombosis.

• Toxicological Research
• /
• v.17
• /
• pp.97-104
• /
• 2001

Three types of proteases (MEF-1, MEF-2 and MEF-3) were purified from the egg cases of Ten-odera sinensis using ammonium sulfate fractionation, gel filtration on Bio-Gel P-60 and affinity chromatography on DEAE Affi-Gel blue gel. The proteases were assessed homogeneous by SDS-polyacrylamide gel electrophoresis and have molecular weight of 31,500, 32,900 and 35,600 Da, respectively. The N-terminal regions of the primary structure were compared and they were found to be different each other. MEFs readily digested the $A\alpha$ - and B$\beta$-chains of fibrinogen and more slowly the ${\gamma}$-chain. The action of the enzymes resulted in extensive hydrolysis of fibrinogen and fibrin, releasing a variety of fibrinopeptides. MEF-1 was inactivated by Cu$^{2+}$ and Zn$^{2+}$ and inhibited by PMSF and chymostatin. MEF-2 was inhibited by PMSF, TLCK. soybean trypsin inhibitor. MEF-3 was only inhibited by PMSF and chymostatin. Antiplasmin was not sensitive to MEF-1 but antithrombin III inhibited the enzymatic activity qf MEF-1. MEF-2 specifically bound to anti plasmin Among the chromogenic protease substrates, the most sensitive one to the hydrolysis of MEFs was benzoyl-Phe-Val-Arg-p-nitroanilide with maximal activity at pH 7.0 and 3$0^{\circ}C$. MEF-1 preferentially cleaved the oxidized B-chain of insulin between Leu15 and Tyr16. In contrast, MEF-2 specifically cleaved the peptide bond between Arg23 and Gly24. D-dimer concentrations increased on incubation of cross-linked fibrin with MEF-1, indicating the enzyme has a strong fibrinolytic activity.ity.

• Journal of Microbiology and Biotechnology
• /
• v.24 no.2
• /
• pp.245-253
• /
• 2014

A fibrinolytic enzyme was produced by an edible mushroom of Pleurotus ostreatus using submerged culture fermentation. The enzyme was purified from the culture supernatant by applying a combination of freeze-thaw treatment, ammonium sulfate precipitation, hydrophobic interaction, and gel filtration chromatographies. The enzyme was purified by a 147-fold, with a yield of 7.54%. The molecular masses of the enzyme an determined by gel filtration and SDS-PAGE were 13.6 and 18.2 kDa, respectively. The isoelectric point of the enzyme was 8.52. It hydrolyzed fibrinogen by cleaving the ${\alpha}$ and ${\beta}$ chains of fibrinogen followed by the ${\gamma}$ chains, and also activated plasminogen into plasmin. The enzyme was optimally active at $45^{\circ}C$ and pH 7.4. The enzyme activity was completely inhibited by EDTA, whereas protease inhibitors of TPCK, SBTI, PMSF, aprotinin and pepstatin showed no inhibition on its activity. The partial amino acid sequences of the enzyme as determined by Q-TOF2 were ATFVGCSATR, GGTLIHESSHFTR, and YTTWFGTFVTSR. These sequences showed a high degree of homology with those of metallo-endopeptidases from P. ostreatus and Armillaria mellea. The purified enzyme can also be applied as a natural agent for oral fibrinolytic therapy or prevention of thrombosis.

• The Journal of Internal Korean Medicine
• /
• v.16 no.1
• /
• pp.197-213
• /
• 1995

The present experiment was desinged to investigate the effects of Sopungtang water extracts on the Cardiovascular System in the Experimental Animals. Thus, the changes of blood pressure and heart rate were measured after oral administration. Measurments of Mortality rate were observed for measuring the effect of Sopungtang water extract. Sopungtang water extract against pulmonary thromboembolism induced by collagen the mixture(0.1ml/10g, 2mg/kg B.W) plus serotonin(5mg/kg B.W) in mouse. The effects of Sopungtang water extract were examined by observing the change of collagen-induced platelet aggregation, coagulation activity, ex vivo and in vitro fibrinolytic activity of euglobulin fraction in rats. The results were summarized as followings. 1. Sopungtang dropped the blood pressure in spontaneous hypertensive rat. 2. The drug increased the auricular blood flow in rabbit. 3. The drug relaxed the artery contraction by pretreated norepinephrine in rat. 4. The drug inhibited the death rate of mouse which was led to thromboembolism by serotonin and collagen. 5. The drug inhibited the platelet aggregation in rat. 6. The drug prolonged the prothrombin time and activated partial thromboplastin time on the test of plasma coagulation factor activity in rat, but was not valuable. 7. The drug increased the antithrombin activity in rat and the fibrinogen lyses time was reduced and lyses area was increased. 8. Sopungtang reduced fibrinogen lyses time of rat in vitro assay. According to the above mentioned results, Sopungtang increased the blood flow and dropped the blood pressure by the dilation of blood vessel. And the drug presented the antithrombin acivity, inhibited the platelet aggregation.