• Reproductive and Developmental Biology
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• 제32권2호
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• pp.89-95
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• 2008

The present study was undertaken to identify changes of plasminogen activators (PAs) in porcine oviductal epithelial cells (POECs) during the estrous cycle classified with post-ovulatory stages (Post-Ov), early to mid-luteal stages (Early-mid L) and pre-ovulatory (Pre-Ov) stages. The urokinase-type plasminogen activator (uPA) was only observed on day 5 and day 7 of culture in the POECs on all the estrous cycles and gradually increased according to increasing culture times, but not Early-mid L. In POECs-conditioned medium, uPA, tissue-type (tPA) and tPA-PA inhibitor (tPA-PAI) activity were observed at all culture times during estrous cycles. The uPA activity of POECs-conditioned medium on Post-Ov stage were significantly (p<0.05) decreased during prolonged cultures. On the other hand, the tPA activity of POECs-conditioned medium at Post-Ov stage was significantly (p<0.05) higher on day 5 than compared to the other days. Although was higher on day 1 at Post-Ov stage, the tPA-PAI activity of POECs-conditioned medium was significantly (p<0.05) higher on day 7 at all stage than that of day 5 of the culture. Taken together, these results suggest that uPA, tPA and tPA-PAI are produced by POECs, and the variations of the PAs activity are regulated in the different stages of the estrous cycle.

• 생약학회지
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• 제29권3호
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• pp.149-154
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• 1998

Binding of urokinase-type plasminogen activator to its cellular receptor accelerate production of plasmin from plasminogen on the cell surface. Plasmin can digest extracellular matrix components and basement membranes through activating certain proMMPs, which is related to the invasiveness to the cells. Plasmin also acts the regulation of blood coagulation and relates closely to cardiovascular diseases such as stroke and coronary occlusion. Therefore, its inhibitors may be useful as antimetastatic agents and to the treatment of cardiovascular diseases. To search for plasmin inhibitors from plant resources, we screened plasmin inhibitory activities with 76 methanol extract of plant species. Among them, three plant samples showed strong inhibitory activities (>70%) and thirteen plant samples showed more than 50% inhibitory activities of plasmin. Their inhibitory activities were not directly related with uPA inhibitory activites and cell viability.

• Radiation Oncology Journal
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• 제24권1호
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• pp.58-66
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• 2006

목적: 방사선에 의한 신장손상은 궁극적으로 신장 섬유화로 인한 신부전으로 나타나며 여기에는 세포외기질의 변화가 동반된다. 방사선 신장손상에서 신사구체 상피세포의 역할을 알아보기 위하여 방사선에 의한 세포외기질과 연관된 여러 유전자 발현의 변화를 알아보고자 하였다. 대상 및 방법 : 백서 사구체 상피세포 (rat glomerular epithelial cell: GEpC) 에 6 MV 선형가속기 (Siemens, USA)를 이용하여 0, 2, 5, 10, 20 Gy 의 단일 방사선량을 조사한 후 각각 6, 24, 48, 72 시간에 시료를 채취하였다. Northern blot, Western blot, Zymography를 이용하여 fibronectin (Fn), plasminogen activator inhibitor-1 (Pai-1), matrix metalloproteinases-2, 9 (MMP-2, 9), tissue inhibitor of matrix metallproteinase-2 (TIMP-2), tissue-type plasminogen activator (t-PA), Urokinase-type plasminogen activator (u-PA)의 발현을 측정하였다. 결과: GEpC 에 대한 10 Gy 단일 방사선 조사후 24 시간부터 Fn mRNA 가 유의한 증가를 나타냈으며 48 시간에 측정한 Fn 단백질은 5, 10 Gy 의 방사선량에서 유의하게 증가되었다. 방사선조사에 의해서 Pai-1 유전자의 발현도 mRNA 및 단백질 단계에서 증가되었으며, 특히 10Gy 조사 후 24, 48 시간에 측정한 mRNA 의 증가는 통계적으로 유의하였다. GEpC에 방사선조사 후 24 시간에 측정한 MMP-2 활성형은 방사선량에 따라 증가하였으나 통계학적 유의성은 없었다. 그밖의 MMP-9, TIMP-2, t-PA 와 u-PA 는 아무런 변화를 나타내지 않았다. 결론 : 방사선에 의하여 GEpC에서 세포외 기질과 관련된 유전자 발현의 번화가 관찰되었으며 이는 방사선 신장 손상에 GEpC가 관여함을 나타낸다.

• 한국동물생명공학회지
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• 제34권3호
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• pp.240-246
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• 2019

The aim of this study was to investigate effect of heat stress on expression levels of plasminogen activators (PAs) related mRNAs and proteins, and changes of PAs activity in porcine endometrial explants. The endometrial explants (200 ± 50 mg) were isolated from middle part of uterine horn at follicular phase (Day 19-21) and were pre-incubated in serum-free culture medium at 38.5℃ in 5% CO₂ for 18 h. Then, the tissues were transferred into fresh medium and were cultured at different temperature (38.5, 39.5, 40.5 or 41.5℃) for 24 h. The expression level of urokinase-type PA (uPA), type-1 PA inhibitor (PAI-1), type-2 PAI (PAI-2), and heat shock protein-90 (HSP-90) mRNA were analysis by reverse-transcription PCR and proteins were measured by western blotting. The supernatant were used for measurement of PAs activity. In results, mRNA and protein levels of HSP-90 was higher in 41.5℃ treatment groups than other treatment groups (p < 0.05). The expression of uPA, PAI-1, and PAI-2 mRNA were slightly increased by heat stress, however, there were no significant difference. Heat stress condition suppressed expression of active uPA and PAI-2 proteins (p < 0.05), whereas PAI-1 protein was increased (p < 0.01). Although PAI-1 protein was increased and active uPA was decreased, PAs activity was greatly enhanced by exposure of heat stress (p < 0.05). These results suggest that heat stress condition could change intrauterine microenvironment through regulation of PAs activity and other factors regarding with activation of PAs might be regulate by heat stress. Therefore, more studies regarding with regulatory mechanism of PAs activation are needed.

• Journal of Gastric Cancer
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• 제4권4호
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• pp.207-212
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• 2004

목적: 고형암이 주위조직으로의 침윤과 타장기로의 전이에는 단백분해효소의 활동이 필요하다. Urokinase type plasminogen activator (uPA)는 serine proteinase의 하나로 세포주의 단백분해와 혈관형성에 중요한 물질로서 고형암의 침윤과 전이에 중요한 역할을 한다. uPA는 유방암, 폐암, 방광암, 위암, 직장결장암, 난소암, 및 연조직 육종에서 중요한 예후지표로 알려져 있다. 본 연구에서 저자들은 위암 조직과 정상 점막에서 uPAR유전자의 발현양상에 대해서 조사하고 기존에 알려진 예후인자와의 연관성을 비교 분석하여 향후 중요한 분자학적 치료 target으로의 역할로서의 의의를 살펴보고자 한다. 대상 및 방법: 1997년 5월 이후 위암으로 진단되어 근치적 혹은 고식적 위장절제술을 시행한 위암 환자 35명을 대상으로 하였다. Northern blot analysis와 RT-PCR을 통해 uPAR mRNA의 발현을 확인하였으며, RNA추출을 위한 재료는 위장절제술 직후 종양조직과 동결절편 생검상 종양세포가 없는 것으로 확인된 부위의 정상 위암 조직을 동일 환자로부터 추출하였다. 이들 환자의 육안적 소견, 조직학적 소견, 생존율 등은 내시경과 수술보고서 병리보고서, 병록지 등을 참고하였다. 결과: uPAR mRNA의 expression과 환자의 예후와의 관계를 평가하기 위해서, gene expression과 이미 확립된 clinicopathologic prognostic와의 관계를 비교하였다. 이러한 factor를 중에서 uPAR mRNA expression은 림프절 전이 여부 (P=0.03), TNM stage (P=0.01)와는 일정한 연관성이 있음을 보였으며, 나이나 성별, 종양크기, histologic type, Lauren classification, Ming classification, serosa invasion, vascular invasion, lymphatic vessel invasion, Neural invasion, omental invasion, macroscopic type와는 상관관계를 보이지 않았다. 결론: 종양 세포에서 uPAR를 평가하는 것은 위암의 예후 뿐만 아니라 질병의 재발을 예측하는데 사용될 수 있을 것으로 생각된다.

• Reproductive and Developmental Biology
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• 제35권3호
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• pp.221-225
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• 2011

This study was undertaken to evaluate the relationship between in vitro maturation and plasminogen activators (PAs) activity on porcine cumulus-oocytes complexes (COCs) exposed to oxidative stress. When COCs were cultured in maturation medium with hydrogen peroxide ($H_2O_2$), the proportion of the germinal vesicle breakdown (GVBD) and oocytes maturation were decrease with addition of $H_2O_2$, and were significantly (p<0.05) lower in medium with 0.1 mM $H_2O_2$ than control group. Also, the rate of degenerated oocytes was increased in as $H_2O_2$ concentration in eased. When COCs were cultured for 48 h, three plasminogen-dependent lytic bands were observed: tissue-type PA (tPA); urokinase-type PA (uPA); and tPA-PA inhibitor (tPA-PAI). PA activity was quantified using SDS-PAGE and zymography. When $H_2O_2$ concentration was increased, tPA and tPA-PAI activities also increased in porcine oocytes cultured for 48 h, but not uPA. In other experiment, embryos were divided into three groups and cultured in (1) control medium, (2) control medium with 1.0 mM $H_2O_2$ and (3) control medium with 1.0 mM $H_2O_2$ along with catalase in concentrations of 0.01, 0.1, and 1.0 mg/ml, respectively. $H_2O_2$ decreased the rate of GVBD and maturation in porcine COCs but catalase revealed protective activity, against oxidative stress caused by $H_2O_2$. In this experiment, tPA and tPA-PAI activities were higher in media with 1.0 mM $H_2O_2$ alone. Increasing concentration of catalase decreased tPA and tPA-PAI activities in porcine oocytes. These results indicate that the exposure of porcine follicular oocytes to ROS inhibits oocytes maturation to metaphase-II stage and increase the oocytes degeneration. Also, we speculated that increased ROS level may trigger tPA and tPA-PAI activities in porcine oocytes matured in vitro.

• 한국수정란이식학회지
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• 제28권3호
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• pp.273-280
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• 2013

The aim of this study was to establish a three dimensional (3D) culture system of endometrial cells and to examine the plasminogen activators (PAs) activity in porcine uterine. The 3D culture system in porcine endometrial cells was composed to mixture 3D gel, stromal cells and epithelial cells. The 3D culture system was used to identify normal structure search as uterine tissue and PAs expression in this study. In results, porcine endometrium epithelial cells forming a top monolayer and endometrium stromal cells developed as fibroblast-like within 3D matrix scaffold. Expression of urokinase-type PA (uPA) and tissue-type PA (tPA) were observed during the 3D culture using immunofluorescence. PA activity in 3D-cultured endometrial cells was no significant difference between the tissue type, but 2D culture system were significantly lower than in 3D-cultured endometrial cells (P<0.05). Therefore, basic system and functional aspect of 3D culture could be established with similar system of endometrium tissue. We suggest that this study was assumed applicable as baseline data to investigate mechanism between porcine uterus cells in vitro.

• Asian Pacific Journal of Cancer Prevention
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• 제13권6호
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• pp.2887-2889
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• 2012

Malignant tumors have a capacity to degrade the extracellular matrix by controlled proteolysis. One system involved in these processes is the urokinase-type plasminogen activator (uPA) system. uPAR levels are elevated in tumors from several types of cancer. Our study was planned to investigate serum and urine levels of uPAR in breast cancer patients (n=180) and healthy controls (n=60) by ELISA. Serum (p<0.001) and urine (p<0.001) uPAR values in the patients were both significantly elevated. High serum and urine levels of uPAR can be used as diagnostic tools in lymph node positive patients.

• BMB Reports
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• 제47권7호
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• pp.388-392
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• 2014

Berberine, a type of isoquinoline alkaloid isolated from Chinese medicinal herbs, has been reported to have various pharmacological activities. Studies have demonstrated that berberine has beneficial effects on vascular remodeling and alleviates restenosis after vascular injury. However, its mechanism of action on vascular smooth muscle cell migration is not fully understood. We therefore investigated the effect of berberine on human aortic smooth muscle cell (HASMC) migration. Boyden chamber assay was performed to show that berberine inhibited HASMC migration dose-dependently. Real-time PCR and Western blotting analyses showed that levels of matrix metalloproteinase (MMP)-2, MMP-9, and urokinase-type plasminogen activator (u-PA) were reduced by berberine at both the mRNA and protein levels. Western blotting assay further confirmed that activities of c-Fos, c-Jun, and NF-${\kappa}B$ were significantly attenuated. These results suggest that berberine effectively inhibited HASMC migration, possibly by down-regulating MMP-2, MMP-9, and u-PA; and interrupting AP-1 and NF-${\kappa}B$ mediated signaling pathways.

• 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.