• Animal cells and systems
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• 제10권2호
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• pp.79-83
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• 2006

Tissue plasminogen activator (tPA) is used to lyse clots and reperfuse brain in ischemic stroke. However, sideeffects of intracerebral hemorrhage (ICH) and edema limit their clinical application. In part, these phenomena has been linked with elevations in matrix metalloproteinase-9 (MMP-9) in neurovascular unit. However little is known about their regulatory signaling pathways in brain cells. Here, I examine the role of MAP kinase pathways in tPA-induced MMP-9 regulation in rat cortical astrocytes. tPA $(1-10\;{\mu}g/ml)$ induced dose-dependent elevations in MMP-9 and MMP-2 in conditioned media. Although tPA increased phosphorylation in two MAP kinases (ERK, JNK), only inhibition of the JNK pathway by the JNK inhibitor SP600126 significantly reduced MMP-9 upregulation. Neither ERK inhibition with U0126 nor p38 inhibition with SB203580 had any significant effects. Taken together, these results suggest that c-jun N-terminal kinase (JNK) plays an essential role for tPA-induced MMP-9 upregulation.

• 한국수정란이식학회지
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• 제25권3호
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• pp.171-177
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• 2010

Plasminogen activators (PAs) are serine proteases that convert plasminogen to plasmin. The PA/plasmin system has been associated with a number of physiological processes such as fibrinolysis, ovulation and fertilization. Although correlations have been reported between reactive oxygen species (ROS) and oocyte maturation, the relationship between PA activity and ROS is unknown. The present study was undertaken to determine the effects of cumulus cells on PA activity in matured porcine oocytes under xanthine (X)-xanthine oxidase (XO) system. When oocytes were matured under the X-XO system, the proportion of oocytes remaining GV stage was higher (p<0.05) in oocytes without cumulus cells. The incidence of degenerated oocytes was higher (p<0.05) in the X+XO ($11.1{\pm}6.1$ and $21.6{\pm}3.4%$) than in the control group ($2.9{\pm}1.8$ and $4.0{\pm}1.6%$). The proportion of TUNEL-positive oocytes and activity of caspase-3 were higher (p<0.05) in cumulus-free oocytes and oocytes exposed to ROS. Tissue-type plasminogen activator-plasminogen activator inhibitor (tPA-PAI) and tissue-type plasminogen activator (tPA) activity were detected in oocytes that were separated from cumulus-oocytes complexs (COCs) at 44 h of maturation culture, and only tPA was produced in oocytes that were denuded before the onset of maturation culture. On the other hand, the activities of PA were increased (p<0.05) when oocytes were cultured under the X-XO system. The higher activity of tPA was observed in denuded oocytes (DOs) underwent apoptotic changes by oxidative stress. In COCs, however, tPA-PAI as well as tPA activity was detected and apoptotic changes such as DNA cleavage or caspase-3 activation were not observed. These results suggest that tP A may be relevant to apoptotic cell death in porcine oocytes by oxidative stress.

• 한국산학기술학회논문지
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• 제18권7호
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• pp.140-148
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• 2017

본 연구는 고혈압전단계 남자 대학생들이 단순 일회성 유산소 운동을 운동강도를 달리하여 혈압, 혈중지질, 섬유소용해 인자에 미치는 영향에 대하여 연구하였다. 총 6명이 최종 실험 참가자로 선정되어 일회성 달리기 운동을 저강도(50-60% 여유심박수(Heart rate reserve(HRR)), 중강도(60-70%HRR), 고강도(70-80%HRR)로 실시하였다. 혈압(수축기와 이완기), 혈중지질(총 콜레스테롤(total cholesterol(TC), 고밀도단백질 콜레스테롤(high-density lipoprotein cholesterol(HDL-C)), 섬유소 용해인자(tissue plasminogen activator(tPA) and plasminogen activator inhibitor-1(PAI-1))는 운동 전과 후 그리고 운동 후 60분 각각의 강도마다 측정되었다. 실험결과는 저강도 그룹의 수축기 혈압은 운동 후에 유의하게 상승하였다(p=0.013). 하지만 중강도와 고강도 그룹의 수축기 혈압은 유의미하지는 않았으나 효과크기로 분석시 운동 후 60분에 -1.33과 -1.23으로 큰 효과를 보이며 낮아졌다. HDL-C는 중강도(p=0.003)와 고강도 그룹(p=0.002)에서 운동 후 60분에 유의하게 증하였다. tPA는 중강(p=0.021)와 고강도 그룹이(p=0.042) 운동 후에 유의하게 증가하였다. 그리하여 일회성 중강도와 고강도 운동을 했을 때 운동 후 저혈압이 나타나고 HDL-C가 높아지고 또한 tPA 농도가 높아져서 고혈압 전 단계 젊은 남자들이 지속적인 중강도 이상의 운동을 하게 되면 고혈압과 심혈관질환을 예방하고 지연시킬 것이다.

• Journal of Ginseng Research
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• 제22권3호
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• pp.216-221
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• 1998

We examined the anti-invasive activity of ginsenosides Rhl, Rha on the highly metastatic HT1080 human fibrosarcoma cell line. In vitro invasion assay showed ginsenoside Rhr reduced tumor cell invasion through a reconstituted basement membrane in a transwell chamber more than ginsenoside Rh1. Significant down-regulation of matrix metalloproteinase-9 (MMP-9) by ginsenosides Rh, and Rh2 was detected by Northern blot analysis. However, the expression of MMP-2 was not affected by Rh, and Rhr. The expression of tissue inhibitor of metalloproteinase-2 (TIMP-2) was increased by Rhl after 0.5, 1 or 3 day-treatment but reduced after 6 day-treatment. However, the expression of TIMP-2 was not changed by treatment with Rh2. Plasminogen activator inhibitor (PAI) and urokinase-type plasmlnogen activator (uPA) were not changed by treatment with Rh1 and Rh2 for 3 and 6 days. Quantitative gelatin-based zymography confirmed a markedly reduced expression of MMP-9 but MMP-2 after treatments with ginsenosides Rhl and Rha. These results suggest that down-regulation of MMP-9 contributes to the anti-invasive activity of ginsenosides Rhl and Rhr in the HT1080 cells.

• 한국식품영양과학회지
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• 제44권4호
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• pp.483-490
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• 2015

본 연구는 본태성 고혈압 쥐인 SHR을 이용하여 복분자 완숙과 에탄올 50% 추출물의 혈압 강하 효과 및 그와 관련된 인자들[endothelial nitric oxide synthase(eNOS), homocysteine, plasminogen activator inhibitor-1(PAI-1), albumin, creatinine, blood lipid 등]에 미치는 영향을 조사하였다. 본태성 고혈압 쥐에게 복분자 완숙과 에탄올 50% 추출물을 12주간 투여한 결과 수축기 혈압이 고혈압대조군에 비하여 약 12% 억제되었다. 또한 eNOS 유전자 발현을 유의하게 증가하였으며 혈청 homocysteine 및 PAI-1의 활성이 눈에 띄게 감소됨을 확인하였다. 또한 심근 호산성 세포수를 감소시켜 심장의 손상 및 병변을 경감시키는 효과가 있음을 확인하였다. 그러나 혈중 지질 및 혈청 albumin, creatinine의 농도에는 각 군 간의 변화가 뚜렷하게 나타나지 않았다. 본 연구 결과 복분자 완숙과 에탄올 50% 추출물은 혈관의 손상을 야기할 수 있는 homocysteine이나 PAI-1과 같은 염증성 인자로부터 혈관을 보호하고 이를 통해 eNOS의 유전자 발현을 개선시킴으로써 혈압의 조절에 영향을 미치는 것으로 나타났으며, 향후 심혈관계 질환의 개선에 도움이 되는 식품소재로서의 가능성이 있음을 확인하였다.

• Bulletin of the Korean Chemical Society
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• 제30권1호
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• pp.77-82
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• 2009

Thrombin activatable fibrinolysis inhibitor (TAFI) also known as plasma procarboxypeptidase B or U is a 60 kD glycoprotein, which is the major modulator of fibrinolysis in plasma. TAFI is a proenzyme, which is activated by proteolytic cleavage to an active carboxypeptidase B-like enzyme (TAFIa, 35.8 kD) by thrombin/thrombomodulin and plasmin. Modulation of fibrinolysis occurs when TAFIa enzymatically removes C-terminal lysine residues of partially degraded fibrin, thereby inhibiting the stimulation of tissue plasminogen activator (t-PA) modulated plasminogen activation. TAFIa undergoes a rapid conformational change at $37{^{\circ}C}$ to an inactive isoform called TAFIai. Potato tuber carboxypetidase inhibitor (PTCI) was shown to specifically bind to TAFIa as well as TAFIai. In this study, a novel immunoassay TAFIa/ai ELISA was used for quantitation of the two TAFI activation isoforms TAFIa and TAFIai. The ELISA utilizes PTCI as the capture agent and a double antibody sandwich technique for the detection. Low levels of TAFIa/ai antigen levels were detected in normal plasma and elevated levels were found in hemophilia A plasmas. TAFIa/ai antigen represents a novel marker to monitor fibrinolysis and TAFIa/ai ELISA may be a valuable assay for studying the role of TAFI in normal hemostasis and in pathological conditions.

• 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.

• 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.

• 생명과학회지
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• 제20권6호
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• pp.838-844
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• 2010

토양에서 생육하는 Streptomyces corcohrussi의 혈전용해효소가 DEAE-Sephadex A-50 그리고 Sephadex G-50 젤 여과를 이용한 크로마토그라피 방법에 의해 순수분리 되었다. SDS-PAGE 분석결과, 분리된 효소는 단일 단백질이고, 그 분자량은 약 34 kDa 이라는 것을 알 수 있었다. 순수분리된 효소의 혈전용해활성은 plasminogen-rich fibrin plate에서 0.8 U/ml 이었으나, plasminogen-free fibrin plate 에서의 그 효소활성은 0.36 U/ml 이하이었다. 이러한 결과로, 순수 분리된 효소가 plasminogen activator 로 작용한다는 것을 알 수 있었다. 단백질 저해제인 $\varepsilon$-ACA, t-AMCHA 와 mercuric chloride의 존재시에 그 혈전용해활성은 24% 이하이었는데, 이러한 결과는 이들 plasmin 저해제 그리고(혹은) fibrinogen을 fibrin으로 전환시키는 과정과 관련된 fibrinogen 저해제에 의해 이 효소가 조절될 수 있음을 나타낼 수 있다. 한편으로, 중금속 이온인 $Zn^{2+}$은 그 활성을 58% 감소시켰다. 순수 분리된 효소의 최적 온도는 약 $50^{\circ}C$ 이었고, 그 효소활성의 92% 이상은 pH 5.0과 8.0 사이에서 유지되었다. 그러므로, 이러한 결과들은 하나의 강력한 혈전용해효소를 제공해서, S. corcohrussi 유래 새로운 혈전용해제의 개발에 기여하도록 한다.

• Biomolecules & Therapeutics
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• 제14권1호
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• pp.30-35
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• 2006

Tissue-type plasminogen activator (t-PA) is a multidomain serine protease containing two kringle domains, TK1-2. Previously, Pichia-derived recombinant human TK1-2 has been reported as an angiogenesis inhibitor although t-PA plays an important role in endothelial and tumor cell invasion. In this work, in order to improve in vivo efficacy of TK1-2 through elimination of immune reactivity, we mutated wild type TK1-2 into non-glycosylated form (NE-TK1-2) and examined whether it retains anti-angiogenic activity. The plasmid expressing NE-TK1-2 was constructed by replacing $Asn^{l17}\;and\;Asn^{184}$ with glutamic acid residues. After expression in Pichia pastoris, the secreted protein was purified from the culture broth using S-sepharose and UNO S1-FPLC column. The mass spectrum of NE-TK1-2 showed closely neighboring two peaks, 19631.87 and 19,835.44 Da, and it migrated as one band in SDS-PAGE. The patterns of CD-spectra of these two proteins were almost identical. Functionally, purified NE-TK1-2 was shown to inhibit endothelial cell migration in response to bFGF stimulation at the almost same level as wild type TK1-2. Therefore, the results suggest that non-glycosylated NETK1-2 can be developed as an effective anti-angiogenic and anti-tumor agent devoid of immune reactivity.