Bone Marrow Mesenchymal Stem Cell-derived Exosomes Promote Plasminogen Activator Inhibitor 1 Expression in Vascular Cells in the Local Microenvironment During Rabbit Osteonecrosis of the Femoral Head

Abstract Background Nontraumatic osteonecrosis of the femoral head (NONFH) is a highly disabling orthopedic disease in young individuals. Plasminogen activator inhibitor 1 (PAI-1) has been reported to be positively associated with NONFH. We aimed to investigate the dysregulating PAI-1 in bone marrow mesenchymal stem cells (BMMSCs) and vascular cells in rabbit steroid-induced NONFH. Methods To verify the hypothesis that BMMSCs could promote thrombus formation in a paracrine manner, we collected exosomes from glucocorticoid-treated BMMSCs (GB-Exo) to determine their regulatory effects on vascular cells. microRNA sequencing was conducted to find potential regulators in GB-Exo. Utilizing gain-of-function and knockdown approaches, we testified regulatory effect of microRNA in exosomes. Results The expression of PAI-1 was significantly increased in local microenvironment of the femoral head in ONFH model. GB-Exo promoted PAI-1 expression in vascular smooth muscle cells and vascular endothelial cells. We also revealed that miR-451-5p in GB-Exo should be responsible for the elevated PAI-1. Moreover, we identified miR-133b-3p and tested its role as an potential inhibitor of PAI-1. Conclusions This study provided considerable evidence for BMMSC exosomal miR-mediated upregulation of the fibrinolytic regulators PAI-1 in vascular cells. The disruption of coagulation and low fibrinolysis in the femoral head will eventually lead to a disturbance in microcirculation of NONFH. We believe that our findings could be of great significance for guiding clinical trials in the future.

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Bone marrow mesenchymal stem cell-derived exosomes promote plasminogen activator inhibitor 1 expression in vascular cells in the local microenvironment during rabbit osteonecrosis of the femoral head

Stem Cell Research & Therapy ◽

10.1186/s13287-020-01991-2 ◽

2020 ◽

Vol 11 (1) ◽

Author(s):

Lu Li ◽

Yikai Wang ◽

Xiaobing Yu ◽

Yongming Bao ◽

Lijia An ◽

...

Keyword(s):

Bone Marrow ◽

Femoral Head ◽

Plasminogen Activator ◽

Vascular Endothelial Cells ◽

Thrombus Formation ◽

Plasminogen Activator Inhibitor ◽

Vascular Cells ◽

Plasminogen Activator Inhibitor 1 ◽

Activator Inhibitor ◽

Pai 1

Abstract Background Nontraumatic osteonecrosis of the femoral head (NONFH) is a highly disabling orthopedic disease in young individuals. Plasminogen activator inhibitor 1 (PAI-1) has been reported to be positively associated with NONFH. We aimed to investigate the dysregulating PAI-1 in bone marrow mesenchymal stem cells (BMMSCs) and vascular cells in rabbit steroid-induced NONFH. Methods To verify the hypothesis that BMMSCs could promote thrombus formation in a paracrine manner, we collected exosomes from glucocorticoid-treated BMMSCs (GB-Exo) to determine their regulatory effects on vascular cells. microRNA sequencing was conducted to find potential regulators in GB-Exo. Utilizing gain-of-function and knockdown approaches, we testified the regulatory effect of microRNA in exosomes. Results The expression of PAI-1 was significantly increased in the local microenvironment of the femoral head in the ONFH model. GB-Exo promoted PAI-1 expression in vascular smooth muscle cells and vascular endothelial cells. We also revealed that miR-451-5p in GB-Exo plays a crucial role for the elevated PAI-1. Moreover, we identified miR-133b-3p and tested its role as a potential inhibitor of PAI-1. Conclusions This study provided considerable evidence for BMMSC exosomal miR-mediated upregulation of the fibrinolytic regulator PAI-1 in vascular cells. The disruption of coagulation and low fibrinolysis in the femoral head will eventually lead to a disturbance in the microcirculation of NONFH. We believe that our findings could be of great significance for guiding clinical trials in the future.

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Bone Marrow Mesenchymal Stem Cell-derived Exosomes Promote Plasminogen Activator Inhibitor 1 Expression in Vascular Cells in the Local Microenvironment During Rabbit Osteonecrosis of the Femoral Head

10.21203/rs.2.23418/v2 ◽

2020 ◽

Author(s):

Lu Li ◽

Yikai Wang ◽

Xiaobing Yu ◽

Yongming Bao ◽

Lijia An ◽

...

Keyword(s):

Bone Marrow ◽

Femoral Head ◽

Plasminogen Activator ◽

Vascular Endothelial Cells ◽

Thrombus Formation ◽

Plasminogen Activator Inhibitor ◽

Vascular Cells ◽

Plasminogen Activator Inhibitor 1 ◽

Activator Inhibitor ◽

Pai 1

Abstract Background: Nontraumatic osteonecrosis of the femoral head (NONFH) is a highly disabling orthopedic disease in young individuals. Plasminogen activator inhibitor 1 (PAI-1) has been reported to be positively associated with NONFH. We aimed to investigate the dysregulating PAI-1 in bone marrow mesenchymal stem cells (BMMSCs) and vascular cells in rabbit steroid-induced NONFH. Methods: To verify the hypothesis that BMMSCs could promote thrombus formation in a paracrine manner, we collected exosomes from glucocorticoid-treated BMMSCs (GB-Exo) to determine their regulatory effects on vascular cells. microRNA sequencing was conducted to find potential regulators in GB-Exo. Utilizing gain-of-function and knockdown approaches, we testified regulatory effect of microRNA in exosomes.Results: The expression of PAI-1 was significantly increased in local microenvironment of the femoral head in ONFH model. GB-Exo promoted PAI-1 expression in vascular smooth muscle cells and vascular endothelial cells. We also revealed that miR-451-5p in GB-Exo plays a crucial role for the elevated PAI-1. Moreover, we identified miR-133b-3p and tested its role as a potential inhibitor of PAI-1. Conclusions: This study provided considerable evidence for BMMSC exosomal miR-mediated upregulation of the fibrinolytic regulators PAI-1 in vascular cells. The disruption of coagulation and low fibrinolysis in the femoral head will eventually lead to a disturbance in microcirculation of NONFH. We believe that our findings could be of great significance for guiding clinical trials in the future.

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Plasminogen Activator Inhibitor-1 Confirms the Diagnosis of Hepatic Veno-Occlusive Disease in Patients With Hyperbilirubinemia After Bone Marrow Transplantation

Blood ◽

10.1182/blood.v89.6.2184 ◽

1997 ◽

Vol 89 (6) ◽

pp. 2184-2188 ◽

Cited By ~ 97

Author(s):

Christoph Salat ◽

Ernst Holler ◽

Hans-Jochem Kolb ◽

Brigitte Reinhardt ◽

Rudolf Pihusch ◽

...

Keyword(s):

Bone Marrow ◽

Bone Marrow Transplantation ◽

Plasminogen Activator ◽

Marrow Transplantation ◽

Hepatic Injury ◽

Plasminogen Activator Inhibitor ◽

Occlusive Disease ◽

Plasminogen Activator Inhibitor 1 ◽

Activator Inhibitor ◽

Pai 1

AbstractHepatic veno-occlusive disease (VOD) is a frequent and severe complication after bone marrow transplantation (BMT). We previously have described plasminogen activator inhibitor-1 (PAI-1) as a possible marker of VOD. To confirm the significance of this finding, we now determined PAI-1 levels in 31 of 186 consecutive patients undergoing BMT who developed hyperbilirubinemia greater than 3 mg/dL for various reasons. Diagnoses were made by clinical criteria and confirmed by biopsy in 23 of 31 patients. They included VOD (n = 7), acute graft-versus-host disease (GVHD) of the liver (n = 7), and other hepatic injury (n = 17). PAI-1 (mean ± SD) was significantly (P < .001) elevated in patients with VOD (321.6 ± 161.2 ng/mL) as compared with patients with GVHD (22.8 ± 8.4 ng/mL) or other hepatic damage (32.8 ± 30.8 ng/mL) at the timepoint of bilirubin increase. At the peak bilirubin concentration, the corresponding PAI-1 levels were 426.1 ± 230.0 ng/mL in patients with VOD, 41.0 ± 20.6 ng/mL in patients with GVHD, and 44.6 ± 32.9 ng/mL in patients with other hepatic injury (P < .001 VOD v GVHD/other hepatic injury). Our results underline the relevance of PAI-1 in the differential diagnosis of hyperbilirubinemia after BMT and its significance as a sensitive and specific marker of severe VOD.

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The Relevance of Plasminogen Activator Inhibitor 1 (PAI-1) as a Marker for the Diagnosis of Hepatic Veno-Occlusive Disease in Patients after Bone Marrow Transplantation

Leukemia & Lymphoma ◽

10.3109/10428199909093722 ◽

1999 ◽

Vol 33 (1-2) ◽

pp. 25-32 ◽

Cited By ~ 15

Author(s):

C. Salat ◽

E. Holler ◽

H J. Kolb ◽

R. Pihusch ◽

B. Reinhardt ◽

...

Keyword(s):

Bone Marrow ◽

Bone Marrow Transplantation ◽

Plasminogen Activator ◽

Marrow Transplantation ◽

Plasminogen Activator Inhibitor ◽

Occlusive Disease ◽

Plasminogen Activator Inhibitor 1 ◽

Activator Inhibitor ◽

Pai 1

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Vascular release of plasminogen activator inhibitor-1 impairs fibrinolysis during acute arterial thrombosis in mice

Blood ◽

10.1182/blood.v96.1.153.013k11_153_160 ◽

2000 ◽

Vol 96 (1) ◽

pp. 153-160

Author(s):

Tomihisa Kawasaki ◽

Mieke Dewerchin ◽

Henri R. Lijnen ◽

Jos Vermylen ◽

Marc F. Hoylaerts

Keyword(s):

Carotid Artery ◽

Plasminogen Activator ◽

Thrombus Formation ◽

Blood Platelets ◽

Plasma Concentrations ◽

Plasminogen Activator Inhibitor ◽

Plasminogen Activator Inhibitor 1 ◽

Deficient Mice ◽

Activator Inhibitor ◽

Pai 1

The role of plasminogen activator inhibitor-1 (PAI-1) in the plasma, blood platelets, and vessel wall during acute arterial thrombus formation was investigated in gene-deficient mice. Photochemically induced thrombosis in the carotid artery was analyzed via transillumination. In comparison to thrombosis in C57BL/6J wild-type (wt) mice (113 ± 19 × 106 arbitrary light units [AU] n = 15, mean ± SEM), thrombosis in PAI-1−/− mice (40 ± 10 × 106 AU, n = 13) was inhibited (P < .01), indicating that PAI-1 controls fibrinolysis during thrombus formation. Systemic administration of murine PAI-1 into PAI-1−/− mice led to a full recovery of thrombotic response. Occurrence of fibrinolytic activity was confirmed in 2-antiplasmin (2-AP)–deficient mice. The sizes of thrombi developing in wt mice, in 2-AP+/− and 2-AP−/− mice were 102 ± 35, 65 ± 8.1, and 13 ± 6.1 × 106 AU, respectively (n = 6 each) (P < .05), compatible with functional plasmin inhibition by 2-AP. In contrast, thrombi in wt mice, t-PA−/− and u-PA−/−mice were comparable, substantiating efficient inhibition of fibrinolysis by the combined PAI-1/2-AP action. Platelet depletion and reconstitution confirmed a normal thrombotic response in wt mice, reconstituted with PAI-1−/− platelets, but weak thrombosis in PAI-1−/− mice reconstituted with wt platelets. Accordingly, murine (wt) PAI-1 levels in platelet lysates and releasates were 0.43 ± 0.09 ng/109 platelets and plasma concentrations equaled 0.73 ± 0.13 ng/mL. After photochemical injury, plasma PAI-1 rose to 2.9 ± 0.7 ng/mL (n = 9, P < .01). The plasma rise was prevented by ligating the carotid artery. Hence, during acute thrombosis, fibrinolysis is efficiently prevented by plasma 2-AP, but also by vascular PAI-1, locally released into the circulation after endothelial injury.

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Down-regulation of Plasminogen Activator Inhibitor 1 Expression Promotes Myocardial Neovascularization by Bone Marrow Progenitors

Journal of Experimental Medicine ◽

10.1084/jem.20040221 ◽

2004 ◽

Vol 200 (12) ◽

pp. 1657-1666 ◽

Cited By ~ 28

Author(s):

Guosheng Xiang ◽

Michael D. Schuster ◽

Tetsunori Seki ◽

Alfred A. Kocher ◽

Shawdee Eshghi ◽

...

Keyword(s):

Bone Marrow ◽

Plasminogen Activator ◽

Plasminogen Activator Inhibitor ◽

Plasminogen Activator Inhibitor 1 ◽

Down Regulation ◽

Type Plasminogen Activator ◽

Urokinase Type Plasminogen Activator ◽

Mrna And Protein Expression ◽

Activator Inhibitor ◽

Pai 1

Human adult bone marrow–derived endothelial progenitors, or angioblasts, induce neovascularization of infarcted myocardium via mechanisms involving both cell surface urokinase-type plasminogen activator, and interactions between β integrins and tissue vitronectin. Because each of these processes is regulated by plasminogen activator inhibitor (PAI)-1, we selectively down-regulated PAI-1 mRNA in the adult heart to examine the effects on postinfarct neovascularization and myocardial function. Sequence-specific catalytic DNA enzymes inhibited rat PAI-1 mRNA and protein expression in peri-infarct endothelium within 48 h of administration, and maintained down-regulation for at least 2 wk. PAI-1 inhibition enhanced vitronectin-dependent transendothelial migration of human bone marrow–derived CD34+ cells, and resulted in a striking augmentation of angioblast-dependent neovascularization. Development of large, thin-walled vessels at the peri-infarct region was accompanied by induction of proliferation and regeneration of endogenous cardiomyocytes and functional cardiac recovery. These results identify a causal relationship between elevated PAI-1 levels and poor outcome in patients with myocardial infarction through mechanisms that directly inhibit bone marrow–dependent neovascularization. Strategies that reduce myocardial PAI-1 expression appear capable of enhancing cardiac neovascularization, regeneration, and functional recovery after ischemic insult.

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Serotonin induces the expression of tissue factor and plasminogen activator inhibitor-1 in cultured rat aortic endothelial cells

Blood ◽

10.1182/blood.v97.6.1697 ◽

2001 ◽

Vol 97 (6) ◽

pp. 1697-1702 ◽

Cited By ~ 37

Author(s):

Hidehiko Kawano ◽

Hajime Tsuji ◽

Hiromi Nishimura ◽

Shinzo Kimura ◽

Shingo Yano ◽

...

Keyword(s):

Endothelial Cells ◽

Receptor Antagonist ◽

Tissue Factor ◽

Plasminogen Activator ◽

Thrombus Formation ◽

Plasminogen Activator Inhibitor ◽

Plasminogen Activator Inhibitor 1 ◽

Aortic Endothelial Cells ◽

Activator Inhibitor ◽

Pai 1

Serotonin (5-hydroxytryptamine, or 5-HT), released from activated platelets, not only accelerates aggregation of platelets but also is known to promote mitosis, migration, and contraction of vascular smooth muscle cells (VSMCs). These effects are considered to contribute to thrombus formation and atherosclerosis. The aim of this study was to investigate the effects of 5-HT on the expressions of coagulative and fibrinolytic factors in rat aortic endothelial cells. Endothelial cells were stimulated with various concentrations of 5-HT (0.1∼10 μM), and the expressions of tissue factor (TF), tissue factor pathway inhibitor (TFPI), plasminogen activator inhibitor-1 (PAI-1), and tissue-type plasminogen activator (TPA) messenger RNAs (mRNAs) were evaluated by Northern blot analysis. The activities of TF and PAI-1 were also measured. TF and PAI-1 mRNA were increased significantly in a concentration- and time-dependent manner. However, TFPI and TPA mRNA expression did not change. The inductions of TF and PAI-1 mRNAs were inhibited by a 5-HT1/5-HT2 receptor antagonist (methiothepin) and a selective 5-HT2A receptor antagonist (MCI-9042). These results indicate that 5-HT increases procoagulant activity and reduces fibrinolytic activities of endothelial cells through the 5-HT2A receptor. It was concluded that the modulation of procoagulant and hypofibrinolytic activities of endothelial cells by 5-HT synergistically promotes thrombus formation at the site of vessel injury with the platelet aggregation, VSMC contraction, and VSMC proliferation.

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Vascular release of plasminogen activator inhibitor-1 impairs fibrinolysis during acute arterial thrombosis in mice

Blood ◽

10.1182/blood.v96.1.153 ◽

2000 ◽

Vol 96 (1) ◽

pp. 153-160 ◽

Cited By ~ 46

Author(s):

Tomihisa Kawasaki ◽

Mieke Dewerchin ◽

Henri R. Lijnen ◽

Jos Vermylen ◽

Marc F. Hoylaerts

Keyword(s):

Carotid Artery ◽

Plasminogen Activator ◽

Thrombus Formation ◽

Blood Platelets ◽

Plasma Concentrations ◽

Plasminogen Activator Inhibitor ◽

Plasminogen Activator Inhibitor 1 ◽

Deficient Mice ◽

Activator Inhibitor ◽

Pai 1

Abstract The role of plasminogen activator inhibitor-1 (PAI-1) in the plasma, blood platelets, and vessel wall during acute arterial thrombus formation was investigated in gene-deficient mice. Photochemically induced thrombosis in the carotid artery was analyzed via transillumination. In comparison to thrombosis in C57BL/6J wild-type (wt) mice (113 ± 19 × 106 arbitrary light units [AU] n = 15, mean ± SEM), thrombosis in PAI-1−/− mice (40 ± 10 × 106 AU, n = 13) was inhibited (P < .01), indicating that PAI-1 controls fibrinolysis during thrombus formation. Systemic administration of murine PAI-1 into PAI-1−/− mice led to a full recovery of thrombotic response. Occurrence of fibrinolytic activity was confirmed in 2-antiplasmin (2-AP)–deficient mice. The sizes of thrombi developing in wt mice, in 2-AP+/− and 2-AP−/− mice were 102 ± 35, 65 ± 8.1, and 13 ± 6.1 × 106 AU, respectively (n = 6 each) (P < .05), compatible with functional plasmin inhibition by 2-AP. In contrast, thrombi in wt mice, t-PA−/− and u-PA−/−mice were comparable, substantiating efficient inhibition of fibrinolysis by the combined PAI-1/2-AP action. Platelet depletion and reconstitution confirmed a normal thrombotic response in wt mice, reconstituted with PAI-1−/− platelets, but weak thrombosis in PAI-1−/− mice reconstituted with wt platelets. Accordingly, murine (wt) PAI-1 levels in platelet lysates and releasates were 0.43 ± 0.09 ng/109 platelets and plasma concentrations equaled 0.73 ± 0.13 ng/mL. After photochemical injury, plasma PAI-1 rose to 2.9 ± 0.7 ng/mL (n = 9, P < .01). The plasma rise was prevented by ligating the carotid artery. Hence, during acute thrombosis, fibrinolysis is efficiently prevented by plasma 2-AP, but also by vascular PAI-1, locally released into the circulation after endothelial injury.

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Bone Marrow Plasminogen Activator Inhibitor-1 Influences the Development of Obesity

Journal of Biological Chemistry ◽

10.1074/jbc.m606214200 ◽

2006 ◽

Vol 281 (43) ◽

pp. 32796-32805 ◽

Cited By ~ 31

Author(s):

Bart M. De Taeye ◽

Tatiana Novitskaya ◽

Linda Gleaves ◽

Joseph W. Covington ◽

Douglas E. Vaughan

Keyword(s):

Bone Marrow ◽

Plasminogen Activator ◽

Visceral Fat ◽

Primary Source ◽

Plasminogen Activator Inhibitor ◽

Plasminogen Activator Inhibitor 1 ◽

Diet Induced Obesity ◽

Activator Inhibitor ◽

Pai 1

Plasma levels of plasminogen activator inhibitor-1 (PAI-1) are elevated in obesity and correlate with body mass index. The increase in PAI-1 associated with obesity likely contributes to increased cardiovascular risk and may predict the development of type 2 diabetes mellitus. Although adipocytes are capable of synthesizing PAI-1, the bulk of evidence indicates that cells residing in the stromal fraction of visceral fat are the primary source of PAI-1. We hypothesized that bone marrow-derived PAI-1, e.g. derived from macrophages located in visceral fat, contributes to the development of diet-induced obesity. To test this hypothesis, male C57BL/6 wild-type mice and C57BL/6 PAI-1 deficient mice were transplanted with either PAI-1-/-, PAI-1+/-, or PAI-1+/+ bone marrow. The transplanted animals were subsequently fed a high fat diet for 24 weeks. Our findings show that only the complete absence of PAI-1 protects from the development of diet-induced obesity, whereas the absence of bone marrow-derived PAI-1 protects against expansion of the visceral fat mass. Remarkably, there is a link between the PAI-1 levels, the degree of inflammation in adipose tissue, and the development of obesity. Based on these findings we suggest that bone marrow-derived PAI-1 has an effect on the development of obesity through its effect on inflammation.

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