Protease Nexin 1 Is a Potent Urinary Plasminogen Activator Inhibitor in the Presence of Collagen Type IV

SummaryWhile recombinant tissue plasminogen activator (rt-PA) is successfully used in human ischemic stroke, it may also cause hemorrhagic complications. Animal experiments have shown that hemorrhages are related to microvascular basal lamina damage. We investigated the effects of different doses of rt-PA on the brain microvasculature. Experimental cerebral ischemia in rats was induced for 3 h and followed by 24 h reperfusion (suture model). Each group of rats (n = 6) received either treatment (0.9, 9, or 18 mg rt-PA/kg body weight) or saline (control group) at the end of ischemia. The loss of microvascular basal lamina antigen collagen type IV was measured by Western blot of the ischemic and non-ischemic basal ganglia and cortex. Compared with the contralateral non-ischemic area, collagen type IV was significantly reduced in the ischemic area: (basal ganglia/cortex) 43% +/- 9% / 64% +/- 4 %. Low/moderate doses of rt-PA had a protective effect: 0.9 mg 79% +/- 3% / 89% +/-6%, 9 mg 72% +/- 9%/ 81% +/- 12% (p < 0.05). Higher doses of rt-PA (18 mg) had a similar effect as seen in untreated controls: 57% +/- 11% / 59% +/- 9% (p < 0.05, Anova). MMP-9 and MMP-2, measured by gelatine zymography, steadily increased over higher doses of rt-PA: MMP-9 (basal ganglia/cortex): control 115% +/- 4% / 123% +/- 3% compared with 18 mg rt-PA 146% +/- 5%/ 162% +/- 6% (p < 0.05) and MMP-2: control 109% +/-4%/ 116% +/- 5% and 18 mg rt-PA 222% +/- 15%/ 252% +/- 2% (p < 0.05). Low to moderate doses of rt-PA protect the microvascular basal lamina, whereas high doses of rt-PA have the opposite effect, probably due to increased coactivation of MMP-2 and MMP-9.

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Rosiglitazone Alone Or in Combination with Tissue Plasminogen Activator Improves Ischemic Brain Injury in an Embolic Model in Rats

Journal of Cerebral Blood Flow & Metabolism ◽

10.1038/jcbfm.2009.87 ◽

2009 ◽

Vol 29 (10) ◽

pp. 1683-1694 ◽

Cited By ~ 29

Author(s):

Chen Xu Wang ◽

Xiuqing Ding ◽

Raza Noor ◽

Christina Pegg ◽

Chunyan He ◽

...

Keyword(s):

Brain Injury ◽

Plasminogen Activator ◽

Collagen Type ◽

Collagen Type Iv ◽

Ischemic Brain Injury ◽

Ischemic Brain ◽

Type Iv ◽

Injured Brain ◽

Tissue Plasminogen ◽

Rosiglitazone Treatment

In this study, we examined whether rosiglitazone, a peroxisome proliferator-activated receptor gamma (PPAR γ) agonist, is neuroprotective in focal ischemic brain injury, and whether rosiglitazone can enhance the protective action of tissue plasminogen activator (tPA), an agent used clinically for thrombolytic therapy. Rats were subjected to ischemic brain injury by embolizing preformed clots into the middle cerebral artery (MCA). Treatment with rosiglitazone reduced infarction and improved functional recovery; it also enhanced the neuroprotective action of tPA and lengthened the time window for initiating tPA treatment. Occlusion of MCA resulted in a loss of collagen type IV, a major structural protein of the microvascular basal lamina, and tPA treatment worsened this loss. Rosiglitazone treatment prevented the reduction of collagen type IV in the ischemic injured brain by inhibiting the activation of matrix metallopeptidase-9 (MMP-9). In addition, rosiglitazone treatment reduced inflammatory reactions in the ischemic injured brain. Rosiglitazone either alone or in combination with tPA is an effective agent in the reduction of ischemic brain injury. The reduction of microvascular damage and inflammation contributes to the beneficial actions of rosiglitazone.

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Matrix metalloproteinase (MMP) induction and inhibition at different doses of recombinant tissue plasminogen activator following experimental stroke

Thrombosis and Haemostasis ◽

10.1160/th07-03-0194 ◽

2007 ◽

Vol 98 (11) ◽

pp. 963-969 ◽

Cited By ~ 11

Author(s):

Dorothe Burggraf ◽

Helge Martens ◽

Martin Dichgans ◽

Gerhard Hamann

Keyword(s):

Matrix Metalloproteinase ◽

Plasminogen Activator ◽

Tissue Plasminogen Activator ◽

Basal Lamina ◽

Collagen Type ◽

Recombinant Tissue Plasminogen Activator ◽

Collagen Type Iv ◽

Type Iv ◽

Tissue Plasminogen ◽

Different Doses

SummaryAlthough recombinant tissue plasminogen activator (rt-PA) is successfully used for thrombolysis in human stroke, it may increase the risk of haemorrhagic complications. It was shown that the matrix metalloproteinase (MMP) system is critically involved in basal lamina degradation after middle cerebral artery occlusion and reperfusion following rt-PA administration. We describe the effects of different doses of rt-PA (saline, 0.9, 9, or 18 mg rt-PA/kg body weight) on the MMPs, their specific inhibitors (TIMPs), and also their inducer protein EMMPRIN following experimental cerebral ischemia (3 hours [h], 24 h reperfusion, suture model) in rats. The amount of MMP-2 and -9 was measured by gelatine zymography, TIMP-1 and -2 by reverse gelatine zymography, and the content of EMMPRIN and the basal lamina component collagen type IV by Western blotting. The amount of both MMPs steadily rose with increasing doses of rt- PA (p<0.05). In contrast, their endogenous inhibitors TIMPs decreased (p<0.001). A balance between the proteases and their inhibitors was achieved at the low dose of 0.9 mg/kg rt-PA in the rats, which significantly coincided with the demonstrated protection of collagen type IV degradation at this dose. The inducer protein EMMPRIN increased in parallel to its substrate MMP-2. Exogenous rt-PA leads to an increase of the MMP-inducing system by EMMPRIN, and a rise of the degrading MMPs follows. However, at low to moderate doses of rt-PA the microvascular basal lamina was protected, probably due to inhibition of MMP-2 and MMP-9 by the upregulation of their inhibitors. This strongly supports use of the lowest effective dosage of rt-PA available.

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Urokinase plasminogen activator independent early experimental thrombus resolution: MMP2 as an alternative mechanism

Thrombosis and Haemostasis ◽

10.1160/th10-03-0184 ◽

2010 ◽

Vol 104 (12) ◽

pp. 1174-1183 ◽

Cited By ~ 24

Author(s):

Vikram Sood ◽

Catherine E. Luke ◽

Joseph Baldwin ◽

Erin M. Miller ◽

Megan Elfline ◽

...

Keyword(s):

Plasminogen Activator ◽

Collagen Type ◽

Vena Cava ◽

Urokinase Plasminogen Activator ◽

Collagen Type Iv ◽

Alternative Mechanism ◽

Compensatory Mechanism ◽

Plasmin Activity ◽

Venous Thrombus ◽

Type Iv

SummaryDeep-vein thrombosis (DVT) resolution is thought to be primarily a urokinase plasminogen activator (uPA) -dependent mechanism, although observations suggest other non-fibrinolytic mechanisms may exist. We explored the role of matrix metalloproteinase (MMP) -2 and –9 in early DVT resolution in uPA-deficient mice. Male B6/SVEV (WT) and genetically matched uPA -/- mice underwent inferior vena cava (IVC) ligation to create stasis venous thrombi, with IVC and thrombus harvest. Thrombus size was similar between WT and uPA -/- mice at day 4, suggesting early non uPA-dependent resolution. Intrathrombus neutrophils and monocytes were reduced 3- and 3.5-fold in uPA -/- mice as compared with WT. By ELISA, tumour necrosis factor α and interleukin 1β were not altered, while interferon (IFN)γ was significantly elevated in uPA -/- mice. A compensatory increase in thrombus tPA was not observed, plasmin activity was reduced and PAI-1 was elevated 2.5-fold in uPA -/- mice. Active MMP2, but not MMP9, was elevated 3-fold in uPA-/- mice as compared with WT as well as MMP-14, an MMP2 activator. Collagen type IV and fibrinogen were reduced in uPA -/- mice thrombi as compared with WT. IFNγ induces MMP2, and blockade of IFNγ was associated with larger venous thrombi and reduced active MMP2, as compared with WT. Consistently, MMP2 -/- mice had larger VT as compared with WT controls, despite normal thrombus plasmin levels. Taken together, early experimental venous thrombus resolution is independent of uPA, and, in part, inflammatory cell influx. MMP2-dependent thrombolysis is an important compensatory mechanism of venous thrombus resolution, possibly by collagen type IV metabolism, and may represent an exploitable therapeutic avenue.

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Reciprocal regulation of urokinase receptor (CD87)-mediated cell adhesion by plasminogen activator inhibitor-1 and protease nexin-1

Journal of Cell Science ◽

10.1242/jcs.00861 ◽

2003 ◽

Vol 117 (3) ◽

pp. 477-485 ◽

Cited By ~ 15

Author(s):

S. M. Kanse

Keyword(s):

Cell Adhesion ◽

Plasminogen Activator ◽

Plasminogen Activator Inhibitor ◽

Urokinase Receptor ◽

Plasminogen Activator Inhibitor 1 ◽

Reciprocal Regulation ◽

Protease Nexin ◽

Activator Inhibitor

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Purification and partial characterization of a plasminogen activator inhibitor from the human glioblastoma, U138

Biochemistry and Cell Biology ◽

10.1139/o88-147 ◽

1988 ◽

Vol 66 (12) ◽

pp. 1270-1277 ◽

Cited By ~ 12

Author(s):

A. Rehemtulla ◽

P. Murphy ◽

M. Dobson ◽

D. A. Hart

Keyword(s):

Plasminogen Activator ◽

Interleukin 1 ◽

Sodium Dodecyl ◽

Biological Response ◽

Apparent Molecular Weight ◽

Plasminogen Activator Inhibitor ◽

Apparent Homogeneity ◽

Inflammatory Stimuli ◽

Protease Nexin ◽

Activator Inhibitor

A plasminogen activator inhibitor was purified to apparent homogeneity from conditioned media of U138 cells. The inhibitor is a glycoprotein with a pI of 5.4 and an apparent molecular weight of 45 000. The inhibitor forms sodium dodecyl sulfate-stable complexes with plasminogen activators and trypsin but not with plasmin, thrombin, or pancreatic kallikrein. Some biochemical and immunochemical characteristics of the U138 inhibitor distinguish it from other known plasminogen activator inhibitors. The expression of this inhibitor by U138 cells could be modulated by incubation in phorbol myristate acetate, interleukin-1, tumor necrosis factor, and γ interferon, but not in β interferon. Thus, the expression of the plasminogen activator inhibitor can be influenced by biological response modifiers known to be active in the brain and in the neural response to inflammatory stimuli. Therefore, this inhibitor, along with protease nexin, may be involved in brain development and regulation.

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Expression of plasminogen activator inhibitor-1 and protease nexin-1 in human astrocytes: Response to injury-related factors

Journal of Neuroscience Research ◽

10.1002/jnr.22412 ◽

2010 ◽

pp. n/a-n/a ◽

Cited By ~ 4

Author(s):

Karin Hultman ◽

Fredrik Blomstrand ◽

Michael Nilsson ◽

Ulrika Wilhelmsson ◽

Kristina Malmgren ◽

...

Keyword(s):

Plasminogen Activator ◽

Plasminogen Activator Inhibitor ◽

Plasminogen Activator Inhibitor 1 ◽

Related Factors ◽

Human Astrocytes ◽

Protease Nexin ◽

Activator Inhibitor

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Urinary Plasminogen Activator Inhibitor-1: A Biomarker of Acute Tubular Injury

American Journal of Nephrology ◽

10.1159/000518455 ◽

2021 ◽

pp. 1-11

Author(s):

María Paniagua-Sancho ◽

Yaremi Quiros ◽

Alfredo G. Casanova ◽

Víctor Blanco-Gozalo ◽

Consuelo Agüeros-Blanco ◽

...

Keyword(s):

Plasminogen Activator ◽

Kidney Injury ◽

Plasminogen Activator Inhibitor ◽

Renal Perfusion ◽

Diagnostic Tools ◽

Plasminogen Activator Inhibitor 1 ◽

Urinary Plasminogen Activator ◽

Activator Inhibitor ◽

Pai 1

Introduction: Acute kidney injury (AKI) is a threatening, multiaetiological syndrome encompassing a variety of forms and damage patterns. AKI lacks sufficiently specific diagnostic tools to evaluate the distinct combination of pathophysiological events underlying each case, which limits personalized and optimized handling. Therefore, a pathophysiological diagnosis based on new urinary biomarkers is sought for practical (readiness and noninvasiveness) and conceptual reasons, as the urine is a direct product of the kidneys. However, biomarkers found in the urine may also have extrarenal origin, thus conveying pathophysiological information from other organs or tissues. Urinary plasminogen activator inhibitor-1 (PAI-1) has been associated to AKI, although its origin and traffic to the urine are not known. Methods: Herein, we studied the blood or renal origin of urinary PAI-1 (uPAI-1) in experimental AKI in Wistar rats, by means of the in situ renal perfusion method. For this purpose, urine was collected while the kidneys of rats with AKI showing increased uPAI-1 excretion, and controls, were in situ perfused with a saline solution. Results: Our results show that during perfusion, PAI-1 remained in the urine of AKI rats, suggesting that renal cells shed this protein directly to the urine. PAI-1 is also significantly increased in the urine of AKI patients. Its low correlation with other urinary markers such as NGAL or NAG suggests that PAI-1 provides complementary and distinct phenotypical information. Conclusion: In conclusion, uPAI-1 is a biomarker produced by damaged kidneys following AKI, whose precise pathophysiological meaning in AKI needs to be further investigated.

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