• 약학회지
• /
• 제39권6호
• /
• pp.666-670
• /
• 1995

The plasmin activity of euglobulin fraction treated with earthworm extract(EWE) increased concentration dependently when EWE(0.5, 1.0 and 2.0 mg each) were added to normal plasma. It was shown that lumbrokinase in EWE may be moved to euglobulin fraction and activated plasminogen. Also anticoagulation and fibbrinolysis were studied when EWE(7.5, 30 and 120 mg/kg) were added to rat model. Prothrombin time (PT) were 19.7${\pm}$ 3.8, 22.5${\pm}$ 2.5 and 24.5${\pm}$ 5.0sec. activated partial thromboplastin time (APTT) were 25.7${\pm}$ 5.9, 28.7${\pm}$ 5.2 and 36.5${\pm}$ 19.1 sec. After 15 days, the production of D-dimer were 242.3${\pm}$ 47.4, 250.0${\pm}$ 11.9 and 205.8${\pm}$ 12.2mg/ml plasma, respectively. These data showed that EWE acted on the coagulation factor of intrinsic and extrinsic system.

• 한국잠사학회:학술대회논문집
• /
• 한국잠사학회 2002년도 Join Meetings of Korean Society of Sericultural Science and Japanse Society of Sericultural Science
• /
• pp.66-66
• /
• 2002

No Abstract, See Full Text

• Bulletin of the Korean Chemical Society
• /
• 제33권9호
• /
• pp.3048-3054
• /
• 2012

The use of aspirin is widely recommended for the prevention of heart attacks owing to its ability to inhibit platelet activation by irreversibly blocking cyclooxygenase 1. However, aspirin also affects the fibrinolytic and hemostatic pathways by mechanisms that are not well understood, causing severe hemorrhagic complications. Here, we investigated the ability of aspirin and aspirin metabolites to inhibit thrombin-activatable fibrinolysis inhibitor (TAFI), the major inhibitor of plasma fibrinolysis. TAFI is activated via proteolytic cleavage by the thrombin-thrombomodulin complex to TAFIa, a carboxypeptidase B-like enzyme. TAFIa modulates fibrinolysis by removing the C-terminal arginine and lysine residues from partially degraded fibrin, which in turn inhibits the binding of plasminogen to fibrin clots. Aspirin and its major metabolites, salicylic acid, gentisic acid, and salicyluric acid, inhibit TAFIa carboxypeptidase activity. Salicyluric acid effectively blocks activation of TAFI by thrombin-thrombomodulin; however, salicylates do not inhibit carboxypeptidase N or pancreatic carboxypeptidase B. Aspirin and other salicylates accelerated the dissolution of fibrin clots and reduced thrombus formation in an in vitro model of fibrinolysis. Inhibition of TAFI represents a novel hemostatic mechanism that contributes to aspirin's therapy-associated antithrombotic activity and hemorrhagic complications.

• BMB Reports
• /
• 제37권5호
• /
• pp.574-581
• /
• 2004

The full-length cDNA of the lumbrokinase fraction 6 (F6) protease gene of Lumbricus rubellus was amplified using an mRNA template, sequenced and expressed in E. coli cells. The F6 protease gene consisted of pro- and mature sequences by gene sequence analysis, and the protease was translated and modified into active mature polypeptide by N-terminal amino acid sequence analysis of the F6 protease. The pro-region of F6 protease consisted of the 44 residues from methionine-1 to lysine-44, and the mature polypeptide sequence (239 amino acid residues and one stop codon; 720 bp) started from isoleucine-45 and continued to the terminal residue. F6 protease gene clones having pro-mature sequence and mature sequence produced inclusion bodies in E. coli cells. When inclusion bodies were orally administrated rats, generated thrombus weight in the rat' venous was reduced by approximately 60% versus controls. When the inclusion bodies were solubilized in pepsin and/or trypsin solutions, the solubilized enzymes showed hemolytic activity in vitro. It was concluded the F6 protease has hemolytic activity, and that it is composed of pro- and mature regions.

• BMB Reports
• /
• 제35권2호
• /
• pp.236-238
• /
• 2002

Based on the zymography analysis, Bacillus sp. DJ-4 (screened from Doen-Jang, a Korean traditional fermented food) secretes seven extracellular fibrinolytic enzymes (EFEs; 68, 64, 55, 45, 33, 27, and 13 kDa) in culture broth. These seven EFEs were analyzed by newly applied SDS-fibrin zymography combined with gradient polyacrylamide (SDS-FZGP). This improved gel system was used with a 5-20% acrylamide gradient in a fibrin zymogram gel for the separation of proteins with molecular masses from below 10kDa to over 100kDa on one gel plate. Using this system, high molecular weight bands (HMWBs) were clearly and sharply resolved. We also examined the relationship between an acrylamide concentration and the enzymatic activity of EFE using densitometric analysis.

• 생약학회지
• /
• 제35권1호통권136호
• /
• pp.52-61
• /
• 2004

Inhibitory activities of 264 methanol extracts, which were prepared from different parts of 210 kinds of wild and medicinal plants, against human thrombin were evaluated. Based on the anti-coagulation activity determined by thrombin time and activated partial thromboplastin time, the 14 extracts were screened. The fibrinolytic activity, heat stability and inhibition of other proteolytic digestive enzymes, such as pepsin, papain, trypsin and chymotrypsin, of the 14 extracts were further determined, and Ginko biloba (herba), Ephedra sinica (radix), Reynoutria elliptica (herba), Amomum tsao-ko Crevost (fructus), and Magnolia officinalis Rehd. et Wils (bark) were finally selected as possible plant sources for anti-thrombosis agent. These results suggested that medicinal and wild plants could be the potential source of thrombin inhibitor.

• 대한한방내과학회지
• /
• 제18권1호
• /
• 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.

• Biomolecules & Therapeutics
• /
• 제3권2호
• /
• pp.165-170
• /
• 1995

The biochemical properties of the fibrinolytic protease of 50,800 Da isolated from the venom of Kgdistrodon blomhoffi brevicaudus were characterized. The enzyme hydrolyzed the carboxyl side of arginine in the synthetic chromogenic peptides, N-Benzoyl-Phe-Val-Arg-pNA and N-p-Tosyl-Gly-Pro-Arg-pNA, and the enzyme activity was inhibited by phenylmethylsulfonylfluoride indicating that the enzyme belongs to the serine protease family. The pretense showed maximum activity at pH 7.5 and inhibited by ZnCl$_2$, CuSO$_4$, but not by soybean trypsin inhibitor, pepstatin A, 2-mercaptoethanol and EDTA. The fm value determined with N-p-Tosyl-Gly-Pro-Arg-pNA was 0.2 mM. The thrombolytic activity of the purified enzyme was evaluated by platelet aggregation test in rabbits. While the platelet count ratio in blood of the rabbits injected with thrombin alone declined from 1.0 to 0.6 within 7 min and maintained around 0.6 for 24 hours thereafter, the ratio rapidly recovered from around 0.6 to 0.8 in 1 hr, to 1.0 in 24 hrs when the rabbits were sequentially treated with thrombin and the purified enzyme. The result showed that the serine protease from A. blomhoffi brevicoudus of 50,800 Da had a thrombolytic activity in vivo and the enzyme might be developed as a therapuetic agent for the treatment of thrombic disease.

• 한국식품영양과학회지
• /
• 제36권7호
• /
• pp.889-895
• /
• 2007

본 연구에서는 홍삼과 청국장을 융합하여 기호도 및 기능적 특성이 강화된 홍삼청국장 개발을 위하여 홍삼엑기스와 홍삼분말 첨가량 및 첨가 방법에 따른 품질특성을 조사하였다. 홍삼엑기스를 첨가하여 청국장을 제조한 결과, 설정된 조건에서는 엑기스의 첨가량이 많을수록 점질물을 생성이 많아지고 환원당 함량과 혈전용해능도 높게 나타났다. 홍삼 제품 규격 사포닌 함량은 홍삼엑기스 4%에서 적합하게 나타났다. 또한 홍삼분말 첨가 청국장은 점질물 생성, 환원당 함량 및 혈전용해능이 엑기스 첨가 청국장과 비슷한 경향으로 나타났고 아미노태질소와 암모니아태질소 함량은 분말 첨가량이 높을수록 점점 감소하는 경향으로 나타났으며 조사포닌 함량은 분말 14% 첨가구가 홍삼성분함유 제품 제조 기준에 적합한 것으로 나타났다. 홍삼첨가 시기를 달리하여 청국장을 제조한 결과, 점질물, 아미노태질소, 암모니아태질소, 환원당 함량 및 혈전용해능은 첨가시기에 따른 차이가 크지 않았으며, 조사포닌 함량은 발효전에 첨가한 구간이 발효후에 첨가한 것보다 더 높은 함량을 나타내었다. 진세노사이드 함량은 첨가시기에 따른 차이는 크지 않았으며 $Rg_{1}$, Re, Rf, $Rh_{1}$, $Rg_{2}$, $Rb_{1}$, Rc, $Rb_{2}$, Rd 및 $Rg_{3}$가 확인되었다. 이상의 결과로 홍삼엑기스 4%를 발효전에 첨가하는 방법이 홍삼융합청국장의 품질특성이 우수하여 최적 조건으로 설정할 수 있었다.

• Journal of Microbiology
• /
• 제45권2호
• /
• pp.158-167
• /
• 2007

In this study, we have cloned a novel cDNA encoding for a papain-family cysteine protease from the Uni-ZAP XR cDNA library of the polychaete, Periserrula leucophryna. This gene was expressed in Escherichia coli using the T7 promoter system, and the protease was characterized after partial purification. First, the partial DNA fragment (498 bp) was amplified from the total RNA via RT-PCR using degenerated primers derived from the conserved region of cysteine protease. The full-length cDNA of cysteine protease (PLCP) was prepared via the screening of the Uni-ZAP XR cDNA library using the $^{32}P-labeled$ partial DNA fragment. As a result, the PLCP gene was determined to consist of a 2591 bp nucleotide sequence (CDS: 173-1024 bp) which encodes for a 283-amino acid polypeptide, which is itself composed of an 59-residue signal sequence, a 6-residue propeptide, a 218-residue mature protein, and a long 3'-noncoding region encompassing 1564 bp. The predicted molecular weights of the preproprotein and the mature protein were calculated as 31.8 kDa and 25 kDa, respectively. The results of sequence analysis and alignment revealed a significant degree of sequence similarity with other eukaryotic cysteine proteases, including the conserved catalytic triad of the $Cys^{90},\;His^{226},\;and\;Asn^{250}$ residues which characterize the C1 family of papain-like cysteine protease. The nucleotide and amino acid sequences of the novel gene were deposited into the GenBank database under the accession numbers, AY390282 and AAR27011, respectively. The results of Northern blot analysis revealed the 2.5 kb size of the transcript and ubiquitous expression throughout the entirety of the body, head, gut, and skin, which suggested that the PLCP may be grouped within the cathepsin F-like proteases. The region encoding for the mature form of the protease was then subcloned into the pT7-7 expression vector following PCR amplification using the designed primers, including the initiation and termination codons. The recombinant cysteine proteases were generated in a range of 6.3 % to 12.5 % of the total cell proteins in the E. coli BL21(DE3) strain for 8 transformants. The results of SDS-PAGE and Western blot analysis indicated that a cysteine protease of approximately 25 kDa (mature form) was generated. The optimal pH and temperature of the enzyme were determined to be approximately 9.5 and $35^{\circ}C$, respectively, thereby indicating that the cysteine protease is a member of the alkaline protease group. The evaluation of substrate specificity indicated that the purified protease was more active towards Arg-X or Lys-X and did not efficiently cleave the substrates with non-polar amino acids at the P1 site. The PLCP evidenced fibrinolytic activity on the plasminogen-free fibrin plate test.