To examine the relationship between plasma plasminogen activator inhibitor-1[PAI-1]antigen concentration and diabetic retinopathy in non-insulin dependent diabetic patients, PAI-1 antigen levels and some fibri-nolytic parameters were studied in 89 non-insulin dependent diabetic patients[mean age 59.8 +/-11.3 years]and 25 normal adults as control[meanage 52.8 +/-14.7 years]. The diabetic patients were classified as three subgroups: no DR[n=34], NPDR[n=29]and PDR[n=26]according to the degree of retinopathy.The PAI-1 antigen concentration was measured by enzyme immunoassay[Innotest PAI Ag kit].The diabetic patients had a significantly higher mean PAI-1 antigen level [34.56 +/-17.80ng/milliliter ]compared to a control group[20.35 +/-15.78 ng/milliliter ][p<0.05].Plasma PAI-1 antigen level was significantly lower in diabetic patients with PDR[27.39 +/-15.54 ng/milliliter ]than in diabetics with no DR[36.87 +/-23.31 ng /milliliter ]or NPDR[39.43 +/-2 0.17 ng/milliliter ][p<0.05], probably because of more extensive systemic endothelial damage. These results support the hypothesis that impaired fibrinolysis due to elevated PAI-1 is associated with the development of retinopathy, and therefore the levels of PAI-1 can be used as useful indicator for the development and progression of proliferative retinopathy.
Adult ; Diabetes Mellitus, Type 2* ; Diabetic Retinopathy* ; Fibrinolysis ; Humans ; Plasma* ; Plasminogen Activator Inhibitor 1* ; Plasminogen Activators

PURPOSE: Renal irradiation can lead to the development of radiation nephropathy, and this is characterized by the accumulation of extracellular matrix and final fibrosis. To determine the possible role of the glomerular epithelial cell, the radiation-induced changes in the expression of its genes associated with the extracellular matrix were analyzed. MATERIALS AND METHODS: Rat glomerular epithelial cells (GEpC) were irradiated with a single dose of 0, 2, 5, 10 and 20 Gy with using 6 MV LINAC (Siemens, USA), and the samples were collected 6, 24, 48 and 72 hours post-irradiation, respectively. Northern blotting, western blotting and zymography were used to measure the expression level of fibronectin (Fn), plasminogen activator inhibitor-1 (Pai-1), matrix metalloproteinases-2, 9 (MMP-2, 9), tissue inhibitor of metalloproteinase-2 (TIMP-2), tissue-type plasminogen activator (t-PA) and urokinase-type plasminogen activator (u-PA). RESULTS: Irradiation with a single dose of 10 Gy resulted in a significant increase in Fn mRNA since 24 hours post-irradiation, and a single dose of 5 and 10 Gy significantly increased the Fn immunoreactive protein measured 48 hours post-irradiation. An increase in Pai-1 mRNA and protein was also observed and especially, a single dose of 10 Gy significantly increased the mRNA measured 24 and 48 hours post-irradiation. The active MMP-2 measured 24 hours post-irradiation slightly increased in a dose dependent manner, but this increase did not reach statistical significance. The levels of MMP-9, TIMP-2, t-PA and u-PA appeared unaltered after irradiation. CONCLUSION: Irradiation of the glomerular epithelial cells altered the expression of genes associated with the extracellular matrix, implying that the glomerular epithelial cell may be involved in the development of radiation nephropathy.
Animals ; Blotting, Northern ; Blotting, Western ; Epithelial Cells* ; Extracellular Matrix ; Fibronectins* ; Fibrosis ; Plasminogen Activator Inhibitor 1* ; Plasminogen Activators ; Rats* ; RNA, Messenger ; Tissue Inhibitor of Metalloproteinase-2 ; Tissue Plasminogen Activator ; Urokinase-Type Plasminogen Activator

BACKGROUND: After multiple trauma, blood coagulation activity is enhanced and fibrinolytic activity is suppressed by overproduction of plasminogen activator inhibitor-1 (PAI-1). Intermittent sequential pneumatic compression (ISPC) is an effective method to prevent deep vein thrombosis. Its action is explained by the mechanical effect on blood flow, as well as by the enhancement of fibrinolysis by the reduction of PAI-1. The aim of this study was to determine the effects of ISPC on coagulation and fibrinolysis after multiple trauma. METHODS: Thirty-nine trauma patients were either treated with ISPC (ISPC group, 20 patients) or without ISPC (control group, 19 patients). We measured the plasma levels of the thrombin antithrombin III complex (TAT), the plasmin alpha 2 plasmin inhibitor complex (PIC), the tissue plasminogen activator (t-PA), and the plasminogen activator inhibitor-1 (PAI-1) on admission and at 1, 2, 3, 6, 12, and 24 hours after admission. RESULTS: The TAT was higher than normal in both groups, with no significant difference between the two groups throughout the study period. The PIC level of ISPC group was significantly higher than that of the control group. In the ISPC group, the PIC level increased gradually, reaching a peak at 3 hours and decreasing thereafter. In the control group, the PIC level increased to a peak level at 2 hours. The TAT/PIC ratio dropped in the first two hours and increased at 3 hours, dropping again thereafter. In the ISPC group, the ratio dropped gradually without an intermittent fluctuation. At 3 and 6 hours, the control group showed a significantly greater ratio compared to the ISPC group. PAI-1 was higher than normal in bothgroups, with a significantly lower level in the ISPC group from 2 hours to 24 hours. For the t-PA level, no difference was noted between the two groups, with the peak level occurring at 1 hour. The PAI-1/t-PA ratio was significantly greater in the control group from 2 hours to 12 hours than in the ISPC group, but the difference was not significant at 24 hours. CONCLUSIONS: In multiple trauma patients, ISPC does not seem to affect coagulation, but enhances fibrinolysis through suppressed PAI-1 production. This effect of ISPC may be maintained for 12 hours.
alpha-2-Antiplasmin ; Antithrombin III ; Blood Coagulation ; Fibrinolysin ; Fibrinolysis* ; Humans ; Multiple Trauma* ; Plasma ; Plasminogen Activator Inhibitor 1 ; Plasminogen Activators ; Thrombin ; Tissue Plasminogen Activator ; Venous Thrombosis

BACKGROUND: After multiple trauma, blood coagulation activity is enhanced and fibrinolytic activity is suppressed. Due to high tissue thromboplastin concentration in cerebral tissue, more serious coagulation and fibrinolytic abnormalities may occur when concomitant head trauma is present. The aim of this study was to determine the changes in coagulation and fibrinolysis after trauma and the effects of head trauma on coagulation and fibrinolysis. METHODS: This study includes 35 trauma patients: 16 patients with head trauma (group A) and 19 patients without head trauma (group B). We measured the plasma levels of functional protein C, antithrombin III (AT III), thrombin antithrombin III complex (TAT), plasmin alpha 2 plasmin inhibitor complex (PIC), tissue plasminogen activator antigen (t-PA), and plasminogen activator inhibitor-1 antigen (PAI-1) on admission and on days 1, 2, 4, and 6 after the trauma. RESULTS: The TAT and the TAT/PIC were significantly higher in group A than in group B on all days. PIC was significantly lower in group A than in group B on all days except the day of admission. Over the course of time, the TAT and the TAT/PIC decreased in both groups and PIC increased. On admission, the PAI-1 of both groups was increased, but it decreased over the course of time. The t-PA was increased on admission, was suppressed on the 1st day, and then increased again. The PAI-1 and the t-PA showed no significant difference between the two groups. CONCLUSIONS: After multiple trauma, coagulation was enhanced and fibrinolysis was suppressed. Enhanced coagulation and suppressed fibrinolysis were significantly greater in group A than in group B.
alpha-2-Antiplasmin ; Antithrombin III ; Blood Coagulation ; Craniocerebral Trauma ; Fibrinolysin ; Fibrinolysis* ; Humans ; Multiple Trauma* ; Plasma ; Plasminogen Activator Inhibitor 1 ; Plasminogen Activators ; Protein C ; Thrombin ; Thromboplastin ; Tissue Plasminogen Activator

OBJECTIVES: Plasma fibrinolytic activity is determined by the balance between plasmonogen activators and their inhibitors. The aim of this study was to compare the fibrinolytic activity before and after exercise of the type 2 diabetic patients with control group. METHODS: We measured plasma tissue-plasminogen activator (t-PA) and plasminogen activator inhibitor-1 (PAI-1) antigen before and after standardized treadmill exercise in 21 type 2 diabetic patients (14 men, 11 women, ages 46.2+/-5.6 years) and 21 sex and age- matched control group (10 men, 11 women, ages 48.6+/-5.4 years). RESULTS: 1) Post exercise t-PA antigen increased in both diabetic group (from 7.36+/-3.89 to 10.62+/-4.81 ng/ml, p<0.05) and control group (from 8.30+/-3.99 to 10.99+/-5.52 ng/ml, p<0.05). But the rise in t-PA antigen with exercise was similar in both group. 2) Both base line and post exercise PAI-1 antigen levels were similar between the diabetic group (from 29.46+/-10.35 to 31.48+/-12.94 ng/ml, p>0.05) and control group (from 30.04+/-10.40 ng/ml to 31.06+/-10.88 ng/ml, p>0.05). 3) In diabetic group, significant correlations between base line PAI-1 antigen levels and serum triglyceride levels were observed. And post exercise PAI-1 antigen levels were correlated with systolic blood pressure. CONCLUSION: The results show that plasma t-PA antigen level is increased after vigorous exercise in patients with type 2 diabetes mellitus and plasma PAI-1 antigen level is not changed. The increment of plasma t-PA level is not different with healthy subjects.
Blood Pressure ; Diabetes Mellitus* ; Diabetes Mellitus, Type 2 ; Female ; Humans ; Male ; Plasma ; Plasminogen Activator Inhibitor 1* ; Plasminogen Activators ; Triglycerides

OBJECTIVETo investigate the association between plasminogen activator inhibitor (PAI)-2 expression and invasive potential in hepatocellular carcinoma (HCC) cells.

METHODSThe HCC cell lines with high, low, and non-metastatic potentials, namely MHCC97-H, MHCC97-L, and SMMC-7721 respectively, were cultured in vitro. Matrigel invasion assay and Western blot of PAI-2 protein expression were conducted.

RESULTSThe number of invaded cells in MHCC97-L was significantly higher than that in SMMC-7721 (P=0.005), whereas that in MHCC97-H was higher than in MHCC97-L (P=0.017) and SMMC-7721 (P=0.001). Contrarily, PAI-2 protein expression was gradually reducing from SMMC-7721, MHCC97-L, to MHCC97-H (MHCC97-H vs. MHCC97-L, P<0.001; MHCC97-H vs. SMMC-7721, P=0.001; MHCC97-L vs. SMMC-7721, P=0.001). The Pearson's correlation analysis revealed a significant negative association between invaded cell number and PAI-2 expression (r=-0.892, P=0.001).

CONCLUSIONPAI-2 expression may be negatively associated with the invasive potential of HCC.

Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Humans ; Liver Neoplasms ; pathology ; Neoplasm Invasiveness ; Plasminogen Activator Inhibitor 2 ; physiology

OBJECTIVEThe purpose of this work was to investigate the distribution pattern of fibrinolytic factors and their inhibitors in rabbit tissues.

METHODSThe components of the fibrinolytic system in extracts from a variety of rabbit tissues, including tissue plasminogen activator (tPA), plasminogen activator inhibitor-1 (PAI-1), plasminogen (Plg), plasmin (Pl) and alpha(2) plasmin inhibitor (alpha(2)PI), were determined by colorimetric assay.

RESULTSThe tissue extracts in renal, small intestine, lung, brain and spleen demonstrated strong fibrinolytic function, in which high activity of tPA, Plg and Pl was manifested; whereas in skeletal muscle, tongue and stomach, higher activity of PAI-1 and alpha(2)PI showed obviously. Also excellent linear correlations were found between levels of tPA and PAI-1, Pl and alpha(2)PI, Plg and Pl. In related tissues, renal cortex and renal marrow showed distinctly higher activity of tPA and lower activity of PAI-1, with the levels of Plg and Pl in renal cortex being higher than those in renal marrow, where the alpha(2)PI level was higher than that in renal cortex. Similarly, the levels of tPA, Plg and Pl in small intestine were higher than those in large intestine, but with respect to PAI-1 and alpha(2)PI, the matter was reverse. In addition, the fibrinolytic activity in muscle tissue was lower, however, the levels of tPA, Plg, and Pl in cardiac muscle were obviously higher than those in skeletal muscles, and the levels of PAI-1 and alpha(2)PI were significantly lower than those in skeletal muscle.

CONCLUSIONOur data demonstrate that a remarkable difference of the fibrinolytic patterns exists in rabbit tissues, which has probable profound significance in understanding the relationship between the function of haemostasis or thrombosis and the physiologic function in tissues.

Animals ; Female ; Fibrinolysin ; metabolism ; Fibrinolysis ; Gastric Mucosa ; metabolism ; Gastrointestinal Tract ; metabolism ; Intestinal Mucosa ; metabolism ; Male ; Organ Specificity ; Plasminogen ; metabolism ; Plasminogen Activator Inhibitor 1 ; metabolism ; Rabbits ; Tissue Extracts ; metabolism ; Tissue Plasminogen Activator ; metabolism ; alpha-2-Antiplasmin ; metabolism

BACKGROUNDRecent studies have proved that brain-derived neurotrophic factor (BDNF) possesses angiogenic activity in vitro and in vivo. However, the proangiogenic mechanism of BDNF has not yet been provided with enough information. To explore the proangiogenic mechanism of BDNF, we investigated the effects of BDNF on extracellular proteolytic enzymes, including matrix metalloproteinases (MMPs) and serine proteases, particularly the urokinase-type plasminogen activator (uPA)-plasmin system in human umbilical vein endothelial cells (HUVECs) model.

METHODSTube formation assay was performed in vitro to evaluate the effects of BDNF on angiogenesis. The HUVECs were treated with various concentrations of BDNF (25 - 400 ng/ml) for different (6 - 48 hours), reverse transcriptase-polymerase chain reaction (RT-PCR) was used to assay MMP-2, MMP-9, TIMP-1, and TIMP-2 mRNA in HUVECs, and the conditioned medium was analyzed for MMP and uPA activity by gelatin zymography and fibrin zymography, respectively. uPA, plasminogen activator inhibitor (PAI)-1, tissue inhibitors of metalloproteinase (TIMP)-1, and TIMP-2 were quantified by western blotting analysis.

RESULTSBDNF elicited robust and elongated angiogeneis in two-dimensional cultures of HUVECs in comparison with control. The stimulation of serum-starved HUVECs with BDNF caused obvious increase in MMP-2 and MMP-9 mRNA expression and induced the pro-MMP-2 and pro-MMP-9 activation without significant differences in proliferation. However, BDNF had no effect on TIMP-1 and TIMP-2 production. BDNF increased uPA and PAI-1 production in a dose-dependent manner. Maximal activation of uPA and PAI-1 expression in HUVECs was induced by 100 ng/ml BDNF, while effects of 200 ng/ml and 400 ng/ml BDNF were slightly reduced in comparison with with those of 100 ng/ml. Protease activity for uPA was also increased by BDNF in a dose-dependent manner. BDNF also stimulated uPA and PAI-1 production beyond that in control cultures in a time-dependent manner from 12 hours to 48 hours after BDNF treatment.

CONCLUSIONSBDNF stimulates MMP and uPA/PAI-1 proteolytic network in HUVECs, which may be important to the acquisition of proangiogenic potential.

Brain-Derived Neurotrophic Factor ; pharmacology ; Cells, Cultured ; Gene Expression Regulation ; drug effects ; Humans ; Matrix Metalloproteinase 2 ; genetics ; Matrix Metalloproteinase 9 ; genetics ; Neovascularization, Physiologic ; drug effects ; Plasminogen Activator Inhibitor 1 ; genetics ; RNA, Messenger ; analysis ; Urokinase-Type Plasminogen Activator ; genetics

OBJECTIVETo study the role of plasminogen activator inhibitor-1 (PAI-1) and tissue plasminogen activator (tPA) in the accumulation of extracellular matrix (ECM) in the kidney of KKAy mice with type 2 diabetes.

METHODSKKAy mice, a type 2 diabetic animal model, and C57BL-J mice were sacrificed at 16, 20, and 24 weeks of age, respectively. The local expression of renal laminin was analyzed with immunohistochemistry. Chromogenic substance was used to show the activity of PAI-1. The mRNA expression of tPA was determined by RT-PCR. The mRNA expression of PAI-1 was measured by reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSLaminnin expression was significantly increased in all age groups of KKAy mice. The tPA mRNA was significantly lower than that in C57BL mice, especially at the age of 16w (only 47%). Otherwise the PAI-1 mRNA expression was remarkably up-regulated than that in C57BL mice.

CONCLUSIONIn type 2 diabetes KKAy mice, the accumulation of ECM may be associated with the abnormal expression of PAI-1/tPA mRNA.

Animals ; Diabetes Mellitus, Experimental ; metabolism ; Diabetes Mellitus, Type 2 ; metabolism ; Extracellular Matrix ; metabolism ; Kidney ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Plasminogen Activator Inhibitor 1 ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Tissue Plasminogen Activator ; biosynthesis ; genetics

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.
Annexin A5* ; Antirheumatic Agents ; Apoptosis ; Auranofin* ; Blotting, Western ; Gene Expression ; Humans* ; Leukemia, Lymphocytic, Chronic, B-Cell ; Plasminogen Activator Inhibitor 2 ; Plasminogen Activators* ; Plasminogen* ; Prostate* ; Prostatic Neoplasms* ; RNA, Small Interfering ; Statistics as Topic