Urokinase 투여가 Euglobulin Fibrinolytic Activity에 미치는 영향

김영선,오도연,홍세용 대한내과학회 1990 대한내과학회지 Vol.38 No.4

Euglobulin fibrinolytic activity was measured during urokinase infusion.(loading dose 200,000IU, maintenance dose 200,000IU/hr for five hours) by a wellcontrolled fibrin plate in seven cases of thrombosis(six cases of cerebral thrombosis and one case of deep vein thrombosis) with regard to the change of the blood fibrinogen PTT, TT and FDP. The basal fibrinolytic activity activity was lower than the normal control in five of the case. The fibrinolytic activity in euglobulin fraction increased during urokinase infusion in all cases, but the range of increase was variable(range: 27.4~8. 1 in case 2, 175.6~69.7 in Case 1). Fibrinolytic activity decreased below the basal level by two hours after the cessation of urokinase infusion in all cases. The basal level of fibrinogen concentration ranged from 335-880 mg/㎗ and decreased after the loading dose of urokinase(200,000IU) in all except case 2. The TT and/or PTT, measured in cases 1,4,6 and 7 related well to the fibrinogen level but was not prolonged more than three times the basal level through the urokinase infusion in all cases. Our results emphasize that an adequate dose of urokinase should be assessed by the fibrinolytic activity itself. If the fibrinolytic activity is evaluated only by a fibrinogen-related coagulation profile such as TT or PTT, the adequate fibrinolytic activity might by measured by the individual variance of the reserve of the fibrinogen synthesis.

Lumbricus Rubellus 가 인체의 섬유소 용해력에 미치는 영향

홍세용(S . Y . Hong),양동호(D . H . Yang),오도연(D . Y . Oh),박선양(S . Y . Park) 대한내과학회 1991 대한내과학회지 Vol.40 No.1

To evaluate the lumbrikinase effect on plasmin (ogen) related physiologic fibrinolytic activity, we measured euglobulin fibrinolytic activity, t-PA activity and t-PA antigen during lumbrikinase therapy and the results were compared with the data of control day of before and after lumbrikinase ingestion. Serial dilution of lumbrikinase showed the fibrinolytic activity on fibrin plate by dose dependent pattern between 375㎍/ml and 1㎍/ml concentration. In regard to the fibrinolytic area, 300㎍ of lunbrikinase equivalent to 25 unit of t-PA and 75㎍ of lumbrikinase equivalent to 5 unit of t-PA. Euglobulin fibrinolytic activity increased in all cases by two or three days after lumbrikinase ingestion but decreased to basal level 36 hours after discontinuation of lunbrikinase. The lumbrikinase added to plasma did not influence to the euglobulin fibrinolytic activity. t-PA ag was increased in 5 out of 6 cases and t-PA activity was increased all of the cases after lumbrikinase ingestion. There was direct correlation between euglobulin fibrinolytic activity and t-PA activity. This finding suggest that the increased fibrinolytic activity of euglobulin after lumbrikinase ingestion is due to increased t-PA rather than direct effect of lumbrikinase itself.

Low Molecular Weight Heparin 이 Fibrinolytic Activity 에 미치는 영향

홍세용(Sae Yong Hong),양동호(Dong Ho Yang) 대한내과학회 1991 대한내과학회지 Vol.40 No.4

To evaluate the influence of standard and low molecular weight heparin (LMWH) on intrinsic and extrinsic fibrinolytic activity, we measured total fibrinolytic activity in dextran sulfate euglobulin fraction, t-PA activity, and t-PA ag in plasma from 10 patients with cerebral infarction before and after heparin injection. The results were compared with that of a control day (on heparin injection day). The increase of total fibrinolytic activity in the 1.MWH group was significantly higher than in the standard heparin group (0. 025 < p < 0.05) and control group ((0.025<p<0,05). The t-PA activity increased in the at 1:00 P.M. samples in 3 cases of the standard heparin group 4 cases of the LMWH group and 4 cases of the saline group. But it decreased in 6 cases of the standard heparin group 6 cases of the LMWH group and 6 cases of the saline group. The change of t-PA activity and the rate of increase and decrease did not differ significantly between the groups (p>0.1 ANOVA). The t-PA ag of the 1:00 P.M. samples increased in 9 cases of the standard heparin group, 4 cases of the LMWH group and 7 cases of the saline group. But it decreased in 1 case of the standard heparin group, 6 cases of the LMWH group and 3 cases of the saline group, The change of t-PA ag and the rate of increase and decrease did not differ significantly between the group (p>0.1 ANOVA). The finding that LMWH in- creased the total fibrinolytic activity without an in- crease of extrinsic fibrinolytic activity suggests that LMWH increases intrinsic fibrinolytic activity.

유전자 cloning에 의한 Bacillus subtilis의 fibrinolytic enzyme 활성 변화

이홍석,유천권,이철수,강상모 한국산업미생물학회 2000 한국미생물·생명공학회지 Vol.28 No.1

구조유전자와 enhancer 유전자를 cloning 하여 fibrinolytic enzyme을 생산하는 B. subtilis K-54와 J-10의 fibrinolytic enzyme의 생산 능력을 높이기 위하여 이 유전자를 포함하는 재조합 vector를 만들어 형질전환하여 그 생산능의 변화를 조사하였다. aprN과 enhancer 유전자인 prtR 유전자의 primer를 제작하여 PCR한 결과, B. subtilis J-10주에서는 aprN과 prtR 유전자 band를 확인하였고 염기서열 분석결과 자체 promoter를 갖는 완전한 유전자임을 확인하여 cloning에 사용하였다. Cloning된 두 유전자를 가지고 B. subtilis J-10과 B. subtilis K-54를 형질전환하기 위해 E. coli/B. subtilis shuttle vector에 cloning하여 재조합 vector를 제작하였다. 그 결과 aprN 유전자를 갖는 pAPR2, prtR 유전자를 갖는 pENC2, 두 유전자 모두 가지는 pFLA1을 만들었으며 B. subtilis J-10과 K-54를 형질전환하여 재조합된 Bacillus속의 균주를 만들어 fibrinolytic activity의 생산량을 조사하였다. pAPR2과 pFLA1로 형질전환하였을 경우 각각 약 27.3 와 16%의 활성 증가를 보였으나 pENC2의 경우 활성 증가를 보이지 않았다. pENC2로 분리주인 B. subtilis K-54를 형질전환하였을 경우 약 5배 정도의 활성 증가를 보여 효소의 생산이 증가하였음을 확인할 수 있었다. The transformation of Bacillus subtilis K-54 and J-10 was carried out with constructed vectors containing structure and enhancer genes of aprN and prtR, to increase their fibrinolytic enzyme activity, Bands for the aprN and prtR genes were indentified from B. subtilis J-10 by PCR that was carried out with the constructed primers for the genes. In addition, the gene fragments contained promoter site based on the results of analysing their nucleotide sequence. The two gene fragments, aprN and prtR, obtained by the PCR, were, then, inserted to vector such as T-vector and E.coli/Bacillus shuttle vector. The constructed vector were designated as pAPR2 (aprN) pENC2(prtR) and pFLA1 (aprN and prtR), respectively. the constructed vector was used for transformation of the strains of B. subtilis J-10 and B. subtilis K-54 and the fibrinolytic activity of the transformed strains was investigated. The introduction of the vector, pAPR2 and pFLA1, resulted in the increase of fibrinolytic enzyme activity in B. subtilis J-10 by 27.3 % and 16 %, respectively. However, the introduction of pENC2 to B. subtilis J-10 did not seem to induce increase of the enzyme activity. The strain of B. subtilis K-54 transformed with pENC2 showed an increased fibrinolytic activity by 5 folds compared with that of the original strain of B. subtilis K-54.

Change of Fibrinolytic and Antioxidative Activities of Codonopsis lanceolata According to Various Storage Conditions, and Heat or Salt Treatments

김준호,--,-- 대한의생명과학회 2005 Biomedical Science Letters Vol.11 No.1

We investigated the changes of fibrinolytic and antioxidative activities of wild or cultivated Codonopsis lanceolata, which were packed in woven polypropylene (WP) film or low density polyethylene (LDPE) film and stored for 0, 15, and 30 days at refrigerated (2~4℃) or room (18~20℃) temperature (So we have 16 samples, and wrote them as the abbreviated words such as W-WP-RE-15, W-WP-RO-15, W-LDPE-RE-15, W-LDPE-RO-15, W-WP-RE-30, W-WP-RO-30, W-LDPE-RE-30, W-LDPE-RO-30, C-WP-RE-15, C-WP-RO-15, C-LDPE-RE-15, C-LDPE-RO-15, C-WP-RE-30, C-WP-RO-30, C-LDPE-RE-30, C-LDPE-RO-30). Fibrinolytic activity of fresh Codonopsis lanceolata cultivated in the mountain or field were 0.8 unit (plasmin unit/ml) or 7.3 units, respectively. In descending order, the activities of wild Codonopsis lanceolata stored for 15 days were as followed ; LDPE-RE (0.70 unit), WP-RO (0.52 unit), WP-RE (0.45 unit), and LDPE-RO (0.30 unit). After 30 days, fibrinolytic activities of them decreased to 0.47 unit (LDPE-RE), 0.28 unit (WP-RO), 0.21 unit (WP-RE), and 0.30 unit (LDPE-RO). Considering from the point of fibrinolytic activity, the optimal storage condition of wild Codonopsis lanceolata was packing with LDPE film and storing at 4℃. The change of fibrinolytic activities of Codonopsis lanceolata cultivated in the field revealed the similar trend as wild samples, but the fibrinolytic activities of 30 days-stored samples were maintained better than the wild Codonopsis lanceolata. Fibinolytic activity of wild Codonopsis lanceolata was increased by heating for 5 min at 100℃ and decreased by addition of NaCl. Antioxidative activities of Codonopsis lanceolata were also compared from the electron donating activity. Fresh Codonopsis lanceolata had about 70% of electron donating activity. Independent of cultivation area, electron donating activity dropped to 19~74% (wild Codonopsis lanceolata) and 27~59% (cultivated Codonopsis lanceolata) during 15 days storage. But after 30 days storage, we obtained the unexpected results, which meant that the activities were higher activities than 15 days-stored samples or even though the fresh samples. In general, Codonopsis lanceolata could maintain antioxidative activities most strongly with LDPE film and chilled condition.

아위버섯 추출물의 혈전용해, 트롬빈저해, 항산화 및 항염증 활성

김은정,김준호 한국버섯학회 2015 한국버섯학회지 Vol.13 No.1

Our study investigated the fibrinolytic, thrombin inhibitory, anti-oxidative and anti-inflammatory activities of the water extract and solvent fractions isolated from Pleurotus ferulea. Fibrinolytic activity was investigated using the fibrin plate method. Thrombin inhibitory activity was used to analyze thrombin inhibitor assay. The DPPH assay was used to estimate antioxidative activity. Inhibition of NO production was measured for anti-inflammatory activity in LPS-activated murine RAW 264.7 macrophage cells. An MTS assay was used to evaluate the effects of the water extract and solvent fractions isolated from Pleurotus ferulea on cell viability. Our results showed the fibrinolytic activity to be strong in the ethyl acetate fraction at 1.33 plasmin units. The ethyl acetate fraction also showed high thrombin inhibitory activity at 94.45%. The anti-oxidative activity of the water extract was 37.01% and the anti-inflammatory activity of the chloroform fraction was 98.13%. These findings suggest that Pleurotus ferulea's extract and fractions could be applicable in the development of functional foods for the treatment and prevention of cardiovascular diseases.

아위버섯 추출물의 혈전용해, 트롬빈저해, 항산화 및 항염증 활성

김은정,김준호 한국버섯학회 2015 한국버섯학회지 Vol.13 No.1

아위버섯을 혈관계질환 예방과 치료를 위한 기능성 식품 개발에 이용하기 위한 기초 자료를 얻기 위해 아위버섯 물추출물과 유기용매 분획물의 혈전용해활성과 트롬빈 저해활성, 항산화활성 및 항염증활성을 확인하였다. 에틸아세테이트 분획물은 1.33 plasmin unit의 높은 혈전용해 활성과, 94.45%의 높은 트롬빈 저해활성을 나타냈으며, 물 추출물은 37.01%의 항산화활성을 나타냈으며, 클로로포름 분획물은 98.13%의 가장 높은 항염증활성을 나타냈다. 실험 결과로부터 아위버섯의 에틸아세테이트 분획물은 높은 혈전용해활성, 트롬빈 저해활성 그리고 클로로포름 분획물은 높은 항염증활성을 나타내 심혈관계 질환 관련 치료나 예방을 위한 기능성식품 개발에 이용 가능할 것으로 기대된다. Our study investigated the fibrinolytic, thrombin inhibitory, anti-oxidative and anti-inflammatory activities of the water extract and solvent fractions isolated from Pleurotus ferulea. Fibrinolytic activity was investigated using the fibrin plate method. Thrombin inhibitory activity was used to analyze thrombin inhibitor assay. The DPPH assay was used to estimate anti-oxidative activity. Inhibition of NO production was measured for anti-inflammatory activity in LPS-activated murine RAW 264.7 macrophage cells. An MTS assay was used to evaluate the effects of the water extract and solvent fractions isolated from Pleurotus ferulea on cell viability. Our results showed the fibrinolytic activity to be strong in the ethyl acetate fraction at 1.33 plasmin units. The ethyl acetate fraction also showed high thrombin inhibitory activity at 94.45%. The anti-oxidative activity of the water extract was 37.01% and the anti-inflammatory activity of the chloroform fraction was 98.13%. These findings suggest that Pleurotus ferulea's extract and fractions could be applicable in the development of functional foods for the treatment and prevention of cardiovascular diseases.

더덕과 일부 한약재 열수추출물의 혼합 비율에 따른 생리활성

오혜숙,김준호 한국지역사회생활과학회 2007 한국지역사회생활과학회지 Vol.18 No.3

The aim of this study was to investigate the physiological functionalities of Codonopsis lanceolata, Glycyrrhiza uralensis, Chaenomeles sinensis, Crataegus pinnafida, and their mixtures. We also determined their antioxidative, fibrinolytic, and αglucosidase inhibitory activities. The antioxidative activities of Codonopsis lanceolata, Glycyrrhiza uralensis, Chaenomeles sinensis, and Crataegus pinnafida were 79%, 88.3%, 89.9%, and 89.3% respectively. Their fibrinolytic activities were 0.80plasmin unit/ml, 0.57 plasmin unit/ml, 0.52 plasmin unit/ml, and 0.53 plasmin unit/ml respectively. The αglucosidase inhibitory activity of Codonopsis lanceolata was 25%. The 10fold diluents of Glycyrrhiza uralensis, Chaenomeles sinensis, and Crataegus pinnafida showed αglucosidase inhibitory activities of 93.6%, 65.3%, and 61.3% respectively. In antioxidative activity tests of the medicinal plants mixtures at various ratios, the mixtures of Glycyrrhiza uralensis, Chaenomeles sinensis, and Crataegus pinnafida with Codonopsis lanceolata showed antioxidative activities of approximately 90%. In fibrinolytic activity tests mixtures(1:1) of Codonopsis lanceolata with Chaenomeles sinensis and Crataegus pinnafida exhibited increases of 23% and 24% in activity respectively. In αglucosidase inhibitory activity tests, a mixture (4:1) of Codonopsis lanceolata and 10fold diluted Glycyrrhiza uralensis showed an inhibitory activity of 98%, a mixture (3:1) of Codonopsis lanceolata and 10fold diluted Chaenomeles sinensis showed an inhibitory activity of 69.6%, and a mixture (1:1) of Codonopsis lanceolata and 10fold diluted Crataegus pinnafida showed an inhibitory activity of 50.2%. In conclusion, the mixtures of Glycyrrhiza uralensis, Chaenomeles sinensis, and Crataegus pinnafida with Codonopsis lanceolata will be used as a material for the development of biofunctional foods.

전통대두발효식품들에서 분리한 Bacillus 균주들의 혈전용해능

이재용,정선주,조민정,조계만,김경민,신정혜,이창권,김정상,김정환 경상대학교 농업생명과학연구원 2014 농업생명과학연구 Vol.48 No.2

The purpose of this study is obtaining strains useful as starters for fermented soyfoods by first isolating strains and then examining the characteristics of each isolate such as antimicrobial activities, fibrinolytic activities, and resistance to NaCl. Through this study, EMD17 and HCD2 strains isolated from fermented soyfoods were identified as B. amyloliquefaciens strains. Both strains possessed very high levels of fibrinolytic activities, which was confirmed by incubation on 4 different media. Previously isolated B. amyloliquefaciens MJ1-4 and B. subtilis W42 possessed very strong antifungal and antibacterial activities, respectively, but their fibrinolytic activities were not known. Through this work, the 2 strains s also possessed fibrinolytic activities. Use of Bacillus strains with antimicrobial activities and improved fibrinolytic activities as starters will help to reduce the contamination by toxinogenic microorganisms and improve the functionalities of foods, eventually contributing to the production of high-quality fermented soyfoods. 본 연구는 전통대두발효식품에서 분리한 Bacillus amyloliquefaciens EMD17, HCD2, 그리고 MJ1-4 3 균주들과 B. subtilis W42 균주의 혈전용해능과 식염저항성을 조사함으로써 대두발효식품 제조에 종균으로 유용한 균주들을 확보하는 것이다. EMD17과 HCD2는 본 연구를 통해 B. amyloliquefaciens 균주들로 동정되었고 우수한 혈전용해능을 지님이 4 종류의 배지에서의 배양을 통해 확인되었다. B. amyloliquefaciens MJ1-4와 B. subtilis W42 균주들은 각각 항진균력과 항균력이 우수한 균주들로 보고된 바 있으나 혈전용해능은 알려지지 않았다. 본 연구를 통해 이들 2 균주도 혈전용해능을 지님이 확인되었다. 항균력과 혈전용해능이 우수한 바실러스 균주들을 종균으로 사용하는 것은 비단 식중독균 오염을 줄여줄 뿐 아니라 혈전용해능과 같은 기능성도 높여줌으로써 궁극적으로 고품질의 대두발효식품 제조에 기여할 것이다.

Fibrinolytic Enzyme Production by Bacillus subtilis KH-4 Isolated from Deonjang

Kim, J.M.,Suh, H.J.,Ahn, S.W.,Kim, M.S.,Oh, S.H. The Korean Society of Food Science and Nutrition 2002 Preventive Nutrition and Food Science Vol.7 No.4

A strong fibrin-specific fibrinolytic enzyme was produced from Bacillus subtilis KH-4 isolated from Deonjang, a Korean fermented soybean paste similar to Japanese miso. The addition of glucose as a carbon source resulted in the highest levels of caseinolytic and fibrinolytic activities. Likewise, the addition of yeast extract as the nitrogen source resulted in the highest caseinolytic and fibrinolytic activities (3473.2 unit and 47.4 munit, respectively), It was observed that out of all metal ion sources only calcium (chloride) enhanced caseinolytic and fibrinolytic activities, with increases of 4949.3 unit and 58.2 unit/mg, respectively. The optimal temperature for the production of the enzyme was found to be 4$0^{\circ}C$ in the optimal medium (glucose 20 g, yeast extract 5 g, CaCl$_2$l g, and NaCl 2 g). The maximum fibrinolytic activity was observed at the late stationary phase. B. subtilis KH-4 produced a fibrinolytic enzyme at 4$0^{\circ}C$, after 30 h growth, which increased up to 54 h and then remained constant. These results suggest that Deonjang has potential as a source of physiologically active anti-thromotic enzymes.