• BMB Reports
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• 제40권2호
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• pp.180-188
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• 2007

In vivo studies have demonstrated that aldosterone is an independent contributor to glomerulosclerosis. In the present study, we have investigated whether aldosterone itself mediated glomerulosclerosis, as angiotensin II (Ang II) did, by inducing cultured renal mesangial cells to produce plasminogen activator inhibitor-1 (PAI-1), and whether these effects were mediated by aldosterone-induced increase in transforming growth factor $\beta_1$ (TGF-$\beta_1$) expression and cellular reactive oxygen species (ROS) activity. Quiescent rat mesangial cells were treated by aldosterone alone or by combination of aldosterone and spironolactone, Ang II, neutralizing antibody to TGF-$\beta_1$ or antioxidant Nacetylcysteme (NAC). This study indicate that aldosterone can increase PAI-1 mRNA and protein expression by cultured mesangial cells alone, which is independent of aldosterone-induced increases in TGF-$\beta_1$ expression and cellular ROS. The effects on PAI-1, TGF-$\beta_1$ and ROS generation were markedly attenuated by spironolactone, a mineralocorticoid receptor antagonist, which demonstrate that mineralocorticoid receptor (MR) may play a role in mediating these effects of aldosterone.

• Journal of Microbiology and Biotechnology
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• 제21권12호
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• pp.1299-1305
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• 2011

An important modification of thrombolytic agents is resistance to plasminogen activator inhibitor-1 (PAI-1). In previous studies, a new truncated PAI-1-resistant variant was developed based on deletion of the first three domains in t-PA and the substitution of KHRR 128-131 amino acids with AAAA in the truncated t-PA. The novel variant expressed in a static culture system of Chinese Hamster Ovary (CHO) DG44 cells exhibited a higher resistance to PAI-1 when compared with the full-length commercial drug; Actylase. In the present study, the truncated-mutant protein was expressed in CHO DG44 cells in 50 ml orbital shaking bioreactors. The final yield of the truncated-mutant in the culture was 752 IU/ml, representing a 63% increase compared with the static culture system. Therefore, these results suggest that using the combined features of a transient and stable expression system is feasible for the production of novel recombinant proteins in the quantities needed for preclinical studies.

• Archives of Pharmacal Research
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• 제21권3호
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• pp.260-265
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• 1998

Angiogenic activity of Aloe vera gel was investigated by in vitro assay. We obtained the most active fraction from dichloromethane extract of Aloe vera gel by partitioning between hexane and 90% aqueous methanol. The most active fraction (F3) increased the proliferation of calf pulmonary artery endothelial (CPAE) cells. In addition, F3 fraction induced CPAE cells to invade type I collagen gel and form capillary-like tube through in vitro angiogenesis assay, and increased the invasion of CPAE cells into matrigel through in vitro invasion assay. Furthermore, the effect on the MRNA expression of proteolytic enzymes which are key participants in the regulation of extracellular matrix degradation was investigated by northern blot analysis. F3 fraction enhanced mRNA expression of urokinase-type plasminogen activator (u-PA), matrix metalloproteinase-2 (MMP-2), and membrane-type MMP (MT-MMP) in CPAE cells whereas the expression of plasminogen activator inhibitory (PAl-1) mRNA was not changed.

• 한국임상수의학회지
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• 제34권4호
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• pp.275-278
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• 2017

An 8-year old female Korean Short Hair cat with a history of paralysis of both hind limbs less than 1 hour before admission was referred. On physical examination, the left hind limb was cold and there was no pulsation or mobility. On abdominal ultrasound examination, a thrombus 8 mm in length was found at the aortic bifurcation. The patient was diagnosed with hypertrophic cardiomyopathy (HCM) and cardiogenic pulmonary edema through radiologic evaluation and echocardiography. A tissue plasminogen activator (tPA) was applied intravenously using an accelerated dosing protocol (1 mg administered intravenously [IV] bolus, 2.5 mg IV over 30 min, 1.5 mg IV over 1 h) to treat the feline arterial thromboembolism. Within 12 h after administration of tPA, pulsation and mobility of both hind limbs were normal, without any noticeable complications. Clopidogrel was prescribed to prevent additional thrombus formation, and pimobendan, benazepril, and furosemide were prescribed for administration at home. The patient was discharged and survived 377 days.

• Preventive Nutrition and Food Science
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• 제9권4호
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• pp.336-341
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• 2004

Leptin, resisitn and PAI-1 (plasminogen activator inhibitor-1) are synthesized and secreted by rodent fat cells and recently postulated to be an important link to obesity. This study was conducted to characterize the hormonal regulation of leptin, resistin, and PAI-1 gene expression in the 3T3-L1 adipocytes. The cells were treated with 0.5 $\mu$M insulin, 1 $\mu$M dexamethasone (Dex), or 0.05 $\mu$M triiodothyronine (T3) for 72 hours. The mRNA levels of each peptide were measured by semi-quantitative RT-PCR. The mRNA level of the leptin-producing ob gene was significantly increased by insulin, Dex, and T3 by 3.2-, 3.1- and 2.7-fold, respectively, compared to the control (p < 0.05). The level of resistin mRNA was increased by insulin, Dex, and T3 by 2.7-, 2.5- and 2-fold, respectively, compared to the control (p < 0.05). Likewise, the level of PAI-1 mRNA was significantly increased by insulin, Dex, and T3 compared to the control (p < 0.05). Taken together, our results suggest that insulin, Dex, and T3 may regulate the gene expression of leptin, resistin, and PAI-1 in 3T3-L1 adipocytes.

• Biomolecules & Therapeutics
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• 제25권2호
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• pp.177-185
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• 2017

Auranofin has been developed as antirheumatic drugs, which is currently under clinical development for the treatment of chronic lymphocytic leukemia. Previous report showed that auranofin induced apoptosis by enhancement of annexin A5 expression in PC-3 cells. To understand the role of annexin A5 in auranofin-mediated apoptosis, we performed microarray data analysis to study annexin A5-controlled gene expression in annexin A5 knockdown PC-3 cells. Of differentially expressed genes, plasminogen activator inhibitor (PAI)-2 was increased by annexin A5 siRNA confirmed by qRT-PCR and western blot. Treatment with auranofin decreased PAI-2 and increased annexin A5 expression as well as promoting apoptosis. Furthermore, auranofin-induced apoptosis was recovered by annexin A5 siRNA but it was promoted by PAI-2 siRNA. Interestingly, knockdown of annexin A5 rescued PAI-2 expression suppressed by auranofin. Taken together, our study suggests that induction of annexin A5 by auranofin may enhance apoptosis through suppression of PAI-2 expression in PC-3 cells.

• Childhood Kidney Diseases
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• 제8권1호
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• pp.26-32
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• 2004

목 적 : 소아 신증후군 환자에서 과응고성 경향을 가지고 있으며 Plasminogen activator inhibitor-1(PAI-1)은 강력하게 섬유소의 용해를 감소시키는 당단백으로 최근 몇몇 연구에 의하면 신증후군에서 증가된 PAI-1과 사구체내의 섬유소원이나 섬유소와 관련된 항체의 침착이 연관되어 있음을 시사하고 있다. 저자는 PAI-1 유전자 다형성 중에서 A-844 G 대립유전자 다형성의 빈도와 유전형을 소아의 신증후군에서 임상경과와의 연관성을 비교 평가하였다. 방 법 : 2001년 10월부터 2003년 1월까지 경희대학교 부속병원 동서신장병연구소에 방문한 146명의 신증후군 환아와 249명의 대조군을 대상으로 하였고 신증훈군 환아는 infrequent relapser(IR)와 frequent relapser(FR)로 다시 나누었다. 이들에 대해 PAI-1 promoter gene의 A-844 G에 대한 중합효소 연쇄반응-제한효소절편길이 다형현상(PCR-RFLP)을 이용하여 유전자형을 A/A, A/G, G/G로 분류하여 비교 분석하였다. 통계는 Graph Pad Prism 통계분석 소프트웨어 version 2.0을 사용하였고 95% 신뢰구간과 P value는 0.05보다 작은 것을 의미 있게 보았다. 결 과 : PAI-1 promoter gene의 A- 844G 다형성의 분포는 대조군애서 G/G 81(32.5%), A/A 42(16.9%), G/A 126(50.6%)이였고 신증후군 그룹 중 IR 그룹은 G/G 29(34.1%), A/A 15(17.7%), G/A 41(48.2%)이였으며 FR 그룹에서는 G/G 17(27.9%), A/A 18(29.5%), G/A 26(42.6%)이었다. 단지 PAI-1 gene의 A-844G의 다형성중 A/A 유전자형이 신증후군 환아 중 FR군에서만 대조군에 비하여 유의하게 증가하였다(16.9% vs 29.5%, OR=2.06, P=0.0251). 반면 A/G 유전자형(OR=0.73, P=0.2639)이나 G/G 유전자형(OR=0.80, P=0.4828)은 통계학적인 의의가 없었고 IR군에서 대조군에 비하여 A/A(OR=1.06, P=0.8690), A/G(OR=0.91, P=0.7063), G/G(OR=1.07, P=0.7880)으로 모두 통계학적으로 유의하지 않았다. 결 론 : A-844G AA 유전자형을 가진 신증후군 환자와 FR와 통계적인 상관관계가 있음을 알수 있었다. 이는 향후 PAI-1 유전자와 과응고성과 관련된 생화학적 검사와의 연관성에 대한 추가적인 조사와 특히, 신증후군의 과응고성과 신증후군 재발과의 연관성이 있는지에 관한 연구가 더 필요할 것으로 사료된다.

• Biomolecules & Therapeutics
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• 제21권2호
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• pp.107-113
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• 2013

Plasminogen activator inhibitor-1 (PAI-1) is a member of serine protease inhibitor family, which regulates the activity of tissue plasminogen activator (tPA). In CNS, tPA/PAI-1 activity is involved in the regulation of a variety of cellular processes such as neuronal development, synaptic plasticity and cell survival. To gain a more insights into the regulatory mechanism modulating tPA/PAI-1 activity in brain, we investigated the effects of proteasome inhibitors on tPA/PAI-1 expression and activity in rat primary astrocytes, the major cell type expressing both tPA and PAI-1. We found that submicromolar concentration of MG132, a cell permeable peptide-aldehyde inhibitor of ubiquitin proteasome pathway selectively upregulates PAI-1 expression. Upregulation of PAI-1 mRNA as well as increased PAI-1 promoter reporter activity suggested that MG132 transcriptionally increased PAI-1 expression. The induction of PAI-1 downregulated tPA activity in rat primary astrocytes. Another proteasome inhibitor lactacystin similarly increased the expression of PAI-1 in rat primary astrocytes. MG132 activated MAPK pathways as well as PI3K/Akt pathways. Inhibitors of these signaling pathways reduced MG132-mediated upregulation of PAI-1 in varying degrees and most prominent effects were observed with SB203580, a p38 MAPK pathway inhibitor. The regulation of tPA/PAI-1 activity by proteasome inhibitor in rat primary astrocytes may underlie the observed CNS effects of MG132 such as neuroprotection.

• 생명과학회지
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• 제19권10호
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• pp.1438-1443
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• 2009

이 연구는 신장섬유아 세포를 이용하여 LPS에 의해 유도된 신장섬유화 표적유전자인 plasminogen activator inhibitor (PAI-1) 발현과 Ascofuranone (AF)에 의한 신장섬유화 저해효과를 연구하였다. 이 연구를 통해 LPS가 PAI-1의 발현을 농도 및 시간 의존적으로 증가시켜 LPS가 신장섬유화 유도물질임을 확인 할 수 있었다. 또한 LPS로 유도된 PAI-1 mRNA 및 단백질 발현 레벨이 AF에 의해 저해되었으며, 신장섬유화의 또 다른 대표유전자인 fibronectin의 단백질 발현도 AF에 의해 억제되어 AF가 신장섬유화를 저해하는 사실을 확인할 수 있었다. 그리고 AF에 대한 PAI-1 프로모터 활성을 조사하기 위하여 p800-PAI-1-luc을 신장섬유아 세포에 형질전환 시킨 결과, AF가 PAI-1의 전사 활성 조절을 통해 발현을 억제한다는 것을 확인하였다. ERK-1/2의 상위에 존재하는 MEK inhibitor를 처리하여 PAI-1의 발현을 확인한 결과에서도 AF를 처리한 경우와 동일하게 PAI-1 발현이 저해되어 LPS로 유도된 PAI-1의 발현이 ERK-1/2에 의해 조절됨을 알 수 있었다. 또한 LPS로 유도된 ERK-1/2의 인산화가 AF 농도의존적으로 저해된 결과는, AF가 ERK-1/2의 활성저해를 통하여 PAI-1 발현을 조절한다는 사실을 확인 할 수 있었다. 따라서 이러한 연구결과 AF가 신장섬유화를 저해하는 유력한 후보물질로서의 가능성을 제시하였다.

• BMB Reports
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• 제47권12호
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• pp.691-696
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• 2014

${\alpha}$-Enolase is a glycolytic enzyme and a surface receptor for plasminogen. ${\alpha}$-Enolase-bound plasminogen promotes tumor cell invasion and cancer metastasis by activating plasmin and consequently degrading the extracellular matrix degradation. Therefore, ${\alpha}$-enolase and plasminogen are novel targets for cancer therapy. We found that the amino acid sequence of a peptide purified from enzymatic hydrolysates of seahorse has striking similarities to that of ${\alpha}$-enolase. In this study, we report that this peptide competes with cellular ${\alpha}$-enolase for plasminogen binding and suppresses urokinase plasminogen activator (uPA)-mediated activation of plasminogen, which results in decreased invasive migration of HT1080 fibrosarcoma cells. In addition, the peptide treatment decreased the expression levels of uPA compared to that of untreated controls. These results provide new insight into the mechanism by which the seahorse-derived peptide suppresses invasive properties of human cancer cells. Our findings suggest that this peptide could emerge as a potential therapeutic agent for cancer.