OBJECTIVETo explore the relationship between phlegm-stasis syndrome (PSS) and the fibrinolytic status in patients with non-alcoholic fatty liver (NAFL).

METHODSSeventy patients with NAFL were divided into the PSS group and non-PSS group according to TCM Syndrome typing, and a control group consisted of 28 healthy subjects was set up. Levels of plasminogen (PLG), tissue plasminogen activator (t-PA), plasminogen activator inhibitor-1 (PAI-1) and D-dimer were determined and compared.

RESULTSThe activity of t-PA in NAFL patients was significantly lower than that in the control group (P<0.05), and PLG and PAI-1 were significantly higher than those in the control group (P<0.05). In respect to the TCM Syndrome typing, in patients of PSS, t-PA was significantly lower and PLG, PAI-1 were significantly higher than those in patients of non-PSS (P<0.05 or P<0.01), while D-dimer was insignificantly different between patients of the two Syndrome types (P>0.05).

CONCLUSIONNAFL patients of PSS type shows significant lower of fibrinolytic activity, indicating that there is certain degree of microcirculatory disturbance and hyper viscosity state, so the application of dissolving phlegm and dispelling stasis principle in treating NAFL is significant.

Adult ; Aged ; Diagnosis, Differential ; Fatty Liver ; blood ; diagnosis ; Female ; Fibrinolysis ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Plasminogen ; metabolism ; Plasminogen Activator Inhibitor 1 ; blood ; Tissue Plasminogen Activator ; blood

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

AIMTo investigate the influences of testosterone with varied concentrations on the functions of HUVEC.

METHODSHuman umbilical vein endothelial cells within 2-3 passages were cultured with testosterone (3 x 10(-10) to 3 x 10(-8), 3 x 10(-6), 3 x 10(-5) mol/ L), and the control confluent cells were cultured in the same medium without steroid. MTT experiment was repeated for 7 days to investigate each groups' cell proliferation. The values of NO were tested as recommended. The tPA and PAI-1 antigen levels were assayed with ELISA Kits.

RESULTSTestosterone at physiologic or lower concentrations (3 x 10(-10) to 3 x 10(-8) mol/L ) had no adverse effect on A490 and NO level, meanwhile, stimulated the secretion of tPA (P < 0.01). However, tPA levels markedly reduced at larger dose (3 x 10(-6) to 3 x 10(-5) mol/L). On the other hand, PAI-1 antigen levels decreased significantly at the testosterone concentrations ranging from 3 x 10(-10) to 3 x 10(-5) mol/L (P < 0.05).

CONCLUSIONTestosterone at physiologically relevant concentrations affectively decreased PAI-1, while increased tPA levels, which suggested that testosterone might have beneficial effects on the Human umbilical vein endothelial cells and cardiovascular system to prevent atherosclerosis.

Cells, Cultured ; Human Umbilical Vein Endothelial Cells ; drug effects ; metabolism ; Humans ; Nitric Oxide ; metabolism ; Plasminogen Activator Inhibitor 1 ; metabolism ; Testosterone ; pharmacology ; Tissue Plasminogen Activator ; metabolism

OBJECTIVETo elucidate how chronic prostatitis affects the expression and activity of the plasminogen activator (PA) system and relates to male infertility.

METHODSTwenty-three normal fertile males and 80 chronic prostatitis patients (40 fertile and 40 infertile) were included in this research. SDS polyacrylamide gel electrophoresis and fibrin overlay method were used to estimate the total PA, and tissue PA (tPA), urokinase type PA (uPA) in semen.

RESULTSTotal PA, tPA and uPA highly expressed in normal males, but decreased in the semen of the chronic prostatitis patients of both the fertile and infertile groups. However, there was no significant difference in total PA between the fertile and infertile patients.

CONCLUSIONChronic prostatitis reduces the secretory function and PA synthesis and secretion of the prostate, but the decrease of PA alone does not cause infertility. PA may be one of the tools for estimating the function of the prostate.

Adult ; Case-Control Studies ; Chronic Disease ; Humans ; Infertility, Male ; metabolism ; Male ; Prostatitis ; metabolism ; Semen ; metabolism ; Tissue Plasminogen Activator ; biosynthesis ; Urokinase-Type Plasminogen Activator ; biosynthesis

OBJECTIVEIn order to investigate the role of shear stress in the regulation of endothelial function, we assessed here effects of shear stress on tissue-type plasminogen activator in human endothelial progenitor cells (EPCs).

METHODSThe peripheral blood mononuclear cells were separated from healthy adult and inducted into EPCs, which were identified by double staining for the fluorescent labeled acetylated-LDL and lectin. EPCs were seeded on the small diameter artificial vessels, and then divided into four different experimental groups including stationary group, low-flow shear stress group (5 dyn/cm(2)), medium-flow shear stress group (15 dyn/cm(2)) and high-flow shear stress group (25 dyn/cm(2)). The levels of t-PA in EPC culture medium at 0 hour, 5 hours, 10 hours, 15 hours, 20 hours and 25 hours after culture were measured by enzyme-linked immunosorbent assay.

RESULTSThe peripheral blood mononuclear cells differentiated into EPCs after induction, which were positively labeled by fluorescent acetylated-LDL and lectin. Shear stress enhanced production of the t-PA by EPCs, which was paralleled to levels and times of shear stress.

CONCLUSIONSShear stress increases t-PA secretion by human EPCs, suggesting that shear stress not only regulates vascular endothelial function but also participates in the pathogenesis of arteriosclerosis.

Adult ; Endothelial Cells ; secretion ; Humans ; Stem Cells ; secretion ; Stress, Mechanical ; Tissue Plasminogen Activator ; metabolism

The present study was performed to investigate the relationship between the concentrations of tissue-type plasminogen activator (t-PA) and plasminogen activator inhibitor (PAI) and the CT images in 23 cases of chronic subdural hematomas (SDHs). The concentrations of t-PA and PAI-1 were quantified by enzyme-linked immunosorbent assay (ELISA). Chronic SDHs were divided into five groups according to their appearance on computed tomography: high-density (n = 4), isodensity (n = 8), low-density (n = 5), mixed-density (n = 3), layering (n = 3) types. The volume of hematoma was measured with an image analyzing software program. The concentrations of t-PA were higher in layering (41.2 +/- 0.3 ng/ml, mean +/- standard error of the mean) and high-density (40.0 +/- 1.1 ng/ml) types compared to those of low-density (23.3 +/- 4.1 ng/ml) and iso-density (25.1 +/- 3.7 ng/ml) types. The concentrations of PAI-1 were lower in layering (95.9 +/- 1.0 ng/ml) and high-density (103.4 +/- 34.5 ng/ml) types compared to that of low-density (192.5 +/- 2.6 ng/ml) type. So the ratio between t-PA and PAI-1 (t-PA/PAI) was greater in layering and high-density types. The volume of hematoma was larger in mixed-density and layering types but statistically insignificant. These results presumably suggest that the ratio between t-PA and PAI concentration may contribute to the pathogenesis of the chronic SDH.
Adult ; Aged ; Enzyme-Linked Immunosorbent Assay ; Female ; Hematoma, Subdural/*metabolism ; Human ; Male ; Middle Age ; Plasminogen Activator Inhibitor 1/*analysis ; Tissue Plasminogen Activator/*analysis ; Tomography, X-Ray Computed

OBJECTIVETo investigate the effect of fibrinogen (Fg), fibrin (Fb) and fibrin (ogen) degradation products (FDPs) on tissue plasminogen activator (tPA) and plasminogen activator inhibitor-1 (PAI-1) expressions of human umbilical vein endothelial cells (HUVECs) in coculture system.

METHODSFg, Fb and FDPs at various concentrations (0, 0.5, 1.5, 3.0, 4.5 and 6.0 g/L) were added to the transwell coculture system of HUVECs and smooth muscle cells (SMCs) for 24 hours. The expressions of tPA and PAI-1 at mRNA level were examined by RT-PCR and tPA and PAI-1 protein and activity were detected by ELISA and substrate chromogenic assays.

RESULTStPA expression was not affected by Fg. Fg at concentrations between 3.0 - 4.5 g/L significantly enhanced the mRNA expression, protein content and activity of PAI-1, while expression of PAI-1 was significantly inhibited by Fg at concentration of 6.0 g/L. Fb at concentrations between 3.0 - 4.5 g/L significantly up-regulated mRNA expression, increased protein content and down-regulated activity of tPA. Fb (1.5 - 4.5 g/L) also enhanced the mRNA expression, increased protein content and activity of PAI-1. FDPs at concentrations 3.0 - 6.0 g/L down-regulated the expression of tPA and FDPs at concentrations 1.5 - 6.0 g/L significantly enhanced PAI-1 mRNA expression.

CONCLUSIONFg, Fb and FDPs play important roles in the pathogenesis of atherosclerosis by modulating the expression of tPA and PAI-1 of endothelial and SMCs.

Animals ; Arteriosclerosis ; metabolism ; pathology ; Cells, Cultured ; Coculture Techniques ; Endothelial Cells ; metabolism ; Fibrin ; metabolism ; Fibrin Fibrinogen Degradation Products ; metabolism ; Fibrinogen ; metabolism ; Humans ; Plasminogen Activator Inhibitor 1 ; metabolism ; RNA, Messenger ; metabolism ; Rabbits ; Tissue Plasminogen Activator ; metabolism

In order to further investigate the effect of annexin II (Ann-II) on tissue plasminogen activator (t-PA)-dependent plasminogen (PLG) activation and its interactive mechanism, recombinant native Ann-II bound t-PA, PLG and plasmin with high affinity was examined. The flow cytometric assay showed that the ann-II expression rate was higher in the human umbilical vein endothelial cell (HUVEC) (87.65%) than in the HL-60 cells as controls (35.79%). Two irrelevant proteins, bovine serum albumin (BSA) and equine IgG (EIG) had no effect on the production of plasmin. Ann-II-mediated enhancement of t-PA-dependent PLG activation was inhibited by epsilon-aminocaproic acid or by pretreatment of Ann-II with carboxypeptidase B with the inhibitive rate being 77.8% and 77.0%, respectively. It was revealed that the effect of Ann-II on PLG activation was specific for t-PA. Urokinase didn't bind to Ann-II, demonstrating the role of receptor-related lysine residues on activation of PLG, showing that the Ann-II-PLG interaction was dependent upon carboxyl-terminal lysine residues. These findings suggest that annexin II-mediated co-assembly of t-PA and PLG may promote plasmin generation and play a key role in modulating fibrinolysis on the endothelial surface.
Annexin A2 ; pharmacology ; Cells, Cultured ; Endothelium, Vascular ; cytology ; metabolism ; Fibrinolysis ; Humans ; Plasminogen ; metabolism ; Recombinant Proteins ; pharmacology ; Tissue Plasminogen Activator ; metabolism ; Umbilical Veins ; cytology

In order to further investigate the effect of annexin II (Ann-II) on tissue plasminogen activator (t-PA)-dependent plasminogen (PLG) activation and its interactive mechanism, recombinant native Ann-II bound t-PA, PLG and plasmin with high affinity was examined. The flow cytometric assay showed that the ann-II expression rate was higher in the human umbilical vein endothelial cell (HUVEC) (87.65%) than in the HL-60 cells as controls (35.79%). Two irrelevant proteins, bovine serum albumin (BSA) and equine IgG (EIG) had no effect on the production of plasmin. Ann-II-mediated enhancement of t-PA-dependent PLG activation was inhibited by epsilon-aminocaproic acid or by pretreatment of Ann-II with carboxypeptidase B with the inhibitive rate being 77.8% and 77.0%, respectively. It was revealed that the effect of Ann-II on PLG activation was specific for t-PA. Urokinase didn't bind to Ann-II, demonstrating the role of receptor-related lysine residues on activation of PLG, showing that the Ann-II-PLG interaction was dependent upon carboxyl-terminal lysine residues. These findings suggest that annexin II-mediated co-assembly of t-PA and PLG may promote plasmin generation and play a key role in modulating fibrinolysis on the endothelial surface.
Annexin A2/*pharmacology ; Cells, Cultured ; Endothelium, Vascular/cytology ; Endothelium, Vascular/*metabolism ; Fibrinolysis ; Plasminogen/*metabolism ; Recombinant Proteins/pharmacology ; Tissue Plasminogen Activator/*metabolism ; Umbilical Veins/cytology

In this study, we assessed the effects of shear stress on the expression of plasminogen activator(tPA and uPA) mRNA in cultured NRK-52E cells (a kidney proximal tubular epithelial cell line of normal rat origin) and investigated the mechanism of tubulointerstitial extracellular matrix (ECM) remodeling in the early stage of diabetic nephropathy (DN). The cultured NRK-52E cells were exposed to shear stress of 5 and 10 dyn/cm2 for 1, 3 and 6 hours respectively. Semi-quantity RT-PCR was used to detect the expression of tPA and uPA mRNA. Shear stress down-regulated the expression of tPA and uPA mRNA in cultured NRK-52E cells in a magnitude and time-dependent way. The results suggested that the increased tubular shear stress in the early-stage of DN could decrease the expression of tPA and uPA in renal proximal tubular cells, lead to the reduction of tubulointerstitial fibrinolytic activity and involve in the remodeling of ECM.
Animals ; Cell Line ; Diabetic Nephropathies ; pathology ; Epithelial Cells ; cytology ; metabolism ; pathology ; Extracellular Matrix ; metabolism ; Humans ; Kidney Tubules, Proximal ; cytology ; metabolism ; pathology ; RNA, Messenger ; genetics ; metabolism ; Rats ; Shear Strength ; Tissue Plasminogen Activator ; genetics ; metabolism ; Urokinase-Type Plasminogen Activator ; genetics ; metabolism