• 생명과학회지
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• 제24권8호
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• pp.873-879
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• 2014

소나무(Pinus densiflora)의 잎의 성분을 분리하여 항트롬빈 활성을 조사하였다. 잎은 70% ethanol로 3회 추출하였으며, 그 추출물은 순차적으로 chloroform과 n-buthanol로 분획하였다. n-buthanol 분획으로부터 얻은 수용성 분획을 MPLC와 HPLC에 의해 분리하였다. 분리한 compound를 $^1H-$과 $^{13}C-NMR$로 동정한 결과 2, 3-dihydro-7-hydroxyl-3-hydroxymethyl-2-(4'-hydroxyl-3-methoxyphenyl)-5-benzofuranpropanol-3-O-${\alpha}$-rhamnopyranoside(a neolignan glycoside)와 2, 3-dihyro-3-hydroxymethyl-7-methoxy-2-(4'-hydroxyphenyl-3'-methoxy)-5-benxofuran propanol 4'-O-${\alpha}$-rhamnopyranoside (icariside $E_4$)의 neolignan 구조임을 확인하였다. 트롬빈 저해활성은 분리된 neolignan glycoside와 icariside $E_4$을 작용하여 혈장안에서 응고시간으로 측정하였다. 그 결과, neolignan glycoside의 응고시간이 icariside $E_4$보다 4배 이상 지연하였다. 저해활성은 neolignan glycoside의 농도가 증가함에 따라 그 시간이 지연되는 것을 확인하였다. 더 나아가 지연기전을 확인하기 위해 thrombin과 순수한 fibrinogen를 반응하였다. 그 결과, 트롬빈과 순수한 피브리노겐의 작용에 의해 응고시간은 지연되었으며, 이는 neolignan glycoside가 피브린형성에 주요한 트롬빈의 활성을 직접 저해하는 것으로 사료된다.

• 생명과학회지
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• 제20권11호
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• pp.1640-1647
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• 2010

혈전성 질환을 예방, 개선할 수 있는 안전한 항혈전제를 개발하기 위해, 일반적으로 항혈전 효과가 있다고 알려진 식용 해조류 35종(갈조류 17종, 홍조류 11종 및 녹조류 7종)을 대상으로 메탄올 추출물을 조제하고, 이들의 thrombin time (TT), prothrombin time (PT), activated partial thromboplastin time (aPTT)을 평가하였다. 트롬빈 저해(TT)의 경우 35종 중에서 감태, 곰피, 대황, 넓패, 패, 알송이모자반 및 야마다모자반에서 강력한 활성을, 프로트롬빈 저해(PT)의 경우 트롬빈 저해활성이 나타난 7종 중 야마다모자반을 제외한 6종에서 강력한 활성을 확인하였으며, 혈액응고인자 저해(aPTT)에서는 감태, 곰피, 대황, 넓패, 패, 알송이모자반 및 톳에서 강력한 활성이 나타났다. 선별된 8종의 시료를 대상으로 다양한 농도에서 항혈전 활성을 평가한 결과 알송이모자반에서 가장 강력한 활성을 확인하였으며, 곰피, 대황 및 넓패가 효과적이었다. 항혈전 활성과 해조류 추출물의 총 flavonoid, 총 polyphenol, 총당 및 환원당 함량과의 상관 관계를 검토한 결과, 활성물질은 flavonoid성 물질로 추측되었으며, 선별된 8종은 총 flavonoid 함량 및 총 polyphenol 함량이 매우 높은 특징을 나타내었다. 본 연구 결과는 경제적이면서 대량공급이 가능한 식용 해조류로부터 신규의 안전한 항혈전제 개발이 가능함을 제시하며, 야마다모자반과 톳은 트롬빈 또는 혈장내 응고 인자에 대한 특이 저해제로 개발 가능함을 제시하고 있다.

• BMB Reports
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• 제46권10호
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• pp.484-489
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• 2013

Piperlonguminine (PL), an important component of Piper longum fruits, is known to exhibit anti-hyperlipidemic, antiplatelet and anti-melanogenic activities. Here, the anticoagulant activities of PL were examined by monitoring activated-partial-thromboplastin-time (aPTT), prothrombin-time (PT), and the activities of thrombin and activated factor X (FXa). The effects of PL on the expressions of plasminogen activator inhibitor type 1 (PAI-1) and tissue-type plasminogen activator (t-PA) were also tested in tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) activated HUVECs. The results showed that PL prolonged aPTT and PT significantly and inhibited the activities of thrombin and FXa. PL inhibited the generation of thrombin and FXa in HUVECs. In accordance with these anticoagulant activities, PL prolonged in vivo bleeding time and inhibited TNF-${\alpha}$ induced PAI-1 production. Furthermore, PAI-1/t-PA ratio was significantly decreased by PL. Collectively, our results suggest that PL possesses antithrombotic activities and that the current study could provide bases for the development of new anticoagulant agents.

• Archives of Pharmacal Research
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• 제24권2호
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• pp.109-113
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• 2001

Curdlan is a natural $\beta$-1,3-glucan produced by Agrobacterium biovar 1. In this study, the anticoagulant activity of sulfoalkyl derivatives of curdlan was investigated by carrying out activated partial thromboplastin time (APTT) assay and compared with that of o-sulfonated curdlan. Approximately 100-fold higher concentration of o-sulfonated curdlan than heparin was required to obtain the same level of the clotting time. Anticoagulant activity of curdlan derivatives was dependent on the degree of sulfation in prolonging the clotting time. However, the chain length of the substituent did not play a role in prolonging the clotting time. The curdlan derivatives enhanced thrombin inhibition by mediating through antithrombin III. The inhibition of thrombin by o-sulfonated curdlan was found to be approximately 10-fold weaker than that by heparin.

• BMB Reports
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• 제32권6호
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• pp.567-572
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• 1999

We have previously shown that a DNA fragment is responsible for the anticoagulatory effect of an earthworm, Lumbricus rubellus. The anticoagluant increased the activated partial thromboplastin time (APTT) and also inhibited the thrombin activity observed with either N-${\alpha}$-p-tosyl-L-arginine methyl ester (TAME) or H-D-phenyl-alanyl-L-pipecoil-L-arginine-p-nitroanilide (S-2238). Since trypsin digestion of the anticoagulant further increased the APTT, the possible presence of a regulatory protein for the anticoagulatory DNA was investigated by digesting the anticoagulant with trypsin and isolating the DNA fragment with C4-reversed phase HPLC. The DNA fragment lacking a regulatory protein was eluted in the flow-through fraction, and analyzed with thrombin and activated factor X. Activated factor X activity was more strongly inhibited than thrombin activity. For DNA digestion, we treated the anticoagulant with DNase and purified the DNA-binding protein with a FPLC Resource-S cation exchange column. The regulatory protein, with an $M_r$ of 55.0 kDa, reduced the anticoagulatory effect of the DNA fragment.

• Archives of Pharmacal Research
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• 제10권2호
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• pp.115-120
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• 1987

Methanol extract of Ilex pubescens roots prolonged bleeding time threefold, and inhibited the generation of malondialdehyde released during platelet aggregation inducted by thrombin. Through several purification procedures, its saponin, named ilexoside, was proved to be responsible for the antithrombotic activities of the plant. Ilexosides A, -D and -J and 24-carboxypomolic acid showed strong inhibitory activities on platelet aggregation induced by thrombin.

• Biomolecules & Therapeutics
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• 제22권3호
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• pp.254-259
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• 2014

The pentacyclic triterpenoid ursolic acid (UA) and its isomer oleanolic acid (OA) are ubiquitous in food and plant medicine, and thus are easily exposed to the population through natural contact or intentional use. Although they have diverse health benefits, reported cardiovascular protective activity is contentious. In this study, the effect of UA and OA on platelet aggregation was examined on the basis that alteration of platelet activity is a potential process contributing to cardiovascular events. Treatment of UA enhanced platelet aggregation induced by thrombin or ADP, which was concentration-dependent in a range of $5-50{\mu}M$. Quite comparable results were obtained with OA, in which OA-treated platelets also exhibited an exaggerated response to either thrombin or ADP. UA treatment potentiated aggregation of whole blood, while OA failed to increase aggregation by thrombin. UA and OA did not affect plasma coagulation assessed by measuring prothrombin time and activated partial thromboplastin time. These results indicate that both UA and OA are capable of making platelets susceptible to aggregatory stimuli, and platelets rather than clotting factors are the primary target of them in proaggregatory activity. These compounds need to be used with caution, especially in the population with a predisposition to cardiovascular events.

• BMB Reports
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• 제48권10호
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• pp.577-582
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• 2015

Cyclopia subternata is a medicinal plant commonly used in traditional medicine to relieve pain. Here, the anticoagulant effects of scolymoside, an active compound in C. subternata, were examined by monitoring activated partial thromboplastin time (aPTT), prothrombin time (PT), and the activities of thrombin and activated factor X (FXa). The effects of scolymoside on plasminogen activator inhibitor type 1 (PAI-1) and tissue-type plasminogen activator (t-PA) expression were evaluated in tumor necrosis factor (TNF)-α-activated human endothelial cells. Treatment with scolymoside resulted in prolonged aPTT and PT and the inhibition of thrombin and FXa activities and production. In addition, scolymoside inhibited thrombin-catalyzed fibrin polymerization and platelet aggregation. Scolymoside also elicited anticoagulant effects in mice, including a significant reduction in the PAI-1 to t-PA ratio. Collectively, these findings indicate that scolymoside possesses anticoagulant activities and could be developed as a novel anticoagulant.

• Korean Journal of Radiology
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• 제22권11호
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• pp.1834-1840
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• 2021

Objective: To analyze the computational fluid dynamics (CFD) of femoral artery pseudoaneurysm (FAP), identify a suitable location and timing for percutaneous thrombin injection (PTI) based on this analysis, and report our clinical experience with the procedure. Materials and Methods: CFD can be used to analyze the hemodynamics of the human body. An analysis using CFD recommended that the suitable location of the needle tip for PTI is at the center of the aneurysm sac and the optimal timing for starting PTI is during the early inflow phase of blood into the sac. Since 2011, seven patients (three male and four female; median age, 60 years [range, 43-75 years]) with FAP were treated with PTI based on the devised suitable location and time. Prior to the procedure, color Doppler ultrasonography was performed to determine the location and timing of the thrombin injection. Results: The technical success rate of the PTI was 100%. The amount of thrombin used for the procedure ranged from 200 IU to 1000 IU (median, 500 IU). None of the patients experienced any symptoms or signs of embolic complications during the procedure. Follow-up CT images did not reveal any embolism in the lower extremities and showed complete thrombosis of the pseudoaneurysm. Conclusion: Based on our study of CFD, PTI administered centrally in the FAP during early inflow, as seen on color Doppler, can be an effective technique.

• 생명과학회지
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• 제18권3호
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• pp.363-368
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• 2008

국내의 자생곤충 76종으로부터 다양한 용매를 이용하여 304종의 추출물을 조제한 후, 이들의 항혈전 활성을 평가하였다. 먼저 트롬빈 저해활성 평가 결과, 방아깨비, 왕잠자리, 비단노린재 및 끝검은메뚜기의 DMSO 추출물 4종, 알락수염노린재, 가시길쭉바구미, 잔날개여치, 털두꺼비하늘소, 송장벌레과 유충 및 등빨간거위벌레의 물 추출물 6종에서 우수한 활성을 확인하였다. 선정된 10종을 대상으로 aPTT 측정결과 비단노린재, 방아깨비 및 잔날개여치 추출물에서 미약한 증가가 나타났으나, 전반적으로 혈전 생성 저해의 유의적인 변화는 인정되지 않아, 이들 추출물들은 트롬빈을 직접 저해하는 것으로 추측되었다. 한편 항트롬빈 활성이 우수한 10종 추출물 중에서 방아깨비, 가시길쭉바구미, 알락수염노린재 및 송장벌레과 유충 추출물에서는 우수한 DPPH 소거능을 나타내어 이들의 약용 곤충자원으로의 개발 가능성을 확인하였다.