EFFECTS OF TGF-β_1 ON THE EXPRESSION OF PLASMINOGEN ACTIVATOR INHIBITOR TYPE 1 IN CULTURED HUMAN RENAL INTERSTITIAL FIBROBLASTS

Objective To investigate the effects of transforming growth factor-β1 (TGF-β1 ) on the expression of plasminogen activator inhibitor type 1 (PAI-1 ) mRNA in renal interstitial fibrosis in vitro. Methods Human renal interstitial fibroblasts were isolated and cultured in vitro. The cells wers stimulated by TGF-β1 with different concentration (0 to 10ng/ml ) at different time (0 to 48h). The expression of PAI-1 mRNA was assayed by RT-PCR. Results TGF-β1, had dose-dependent and time-dependent effects on the expression of PAI-1 mRNA in renal interstitial fibroblasts. Conclusion TGF-β1 may partic- ipate in renal fibrosis with inducing the expression of PAI-1 mRNA in renal fibroblasts and affecting the synthesis and degradation of extracellular matrix (ECM).

Idiopathic pulmonary fibrosis in relation to gene polymorphisms of transforming growth factor-β1 and plasminogen activator inhibitor 1

Background Idiopathic pulmonary fibrosis （IPF） is a progressive and lethal fibrotic lung disease of unknown etiology. Host susceptibility or genetic factors may be important for the predisposition to it. Transforming growth factor-β1 （TGF-β1 a potent profibrotic cytokine） and plasminogen activator inhibitor 1 （PAl-1） play important roles in the development of pulmonary fibrosis. The objective of the study was to investigate the association between the gene polymorphisms of TGF-β1 869 T〉C and PAl-1 4G/5G and the susceptibility to IPF in Han ethnicity. Methods Polymerase chain reaction （PCR） and restriction fragment length polymorphism were performed to analyse the gene polymorphisms of TGF-β1 in 869T〉C and PAl-1 4G/5G in 85 IPF patients and 85 healthy controls matched in age, gender, race and smoker status. Results There was a significant difference in 869T〉C genotype distribution of TGF-β1 between IPF cases and controls, a significant negative association between TC genotype and the development of IPF （OR=0.508, 95% CI： 0.275-0.941） and a positive association between CC genotype and the development of IPF （OR=1.967, 95% CI： 1.063-3.641）. There was a significant positive association between PAl-1 5G/5G genotype and the development of IPF （OR=0.418, 95% CI： 0.193-0.904）. Conclusions Gene polymorphisms of TGF-β1 in 869T〉Cand PAl-1 4G/5G may affect the susceptibility to IPF in Han ethnicity. Further investigations are needed to confirm these findings and assess their biological significance in the development of the disease in this ethnic population.

Activation of peroxisome proliferator-activated receptor α in human endothelial cells increases plasminogen activator inhibitor type-1 expression

Objective To investigate the effect of peroxisome proliferator-activated receptors (PPARs) activators on plasminogen activator inhibitor 1 (PAI-1) expression in human umbilical vein endothelial cells and elucidate a possible mechanism. Methods Human umbilical vein endothelial cells (HUVECs) were obtained from normal fetus, and cultured conventionally. Then the HUVEC were exposed to fatty acids and prostaglandin J 2 in varying concentrations with fresh media. RT-PCR and ELISA were used to determine the expression of PPAR and PAI-1 in HUVECs. Transient co-transfection of PAI-1 promoter and PPARα gene or PPARγ gene to ECV304 was performed.Results PPARα, PPARδ and PPARγ mRNA in HUVECs were detected by RT-PCR. Treatment of HUVECs with PPARα and PPARγ activators-linolenic acid, linoleic acid, oleic acid and prostaglandin J 2, but not with stearic acid could augment PAI-I mRNA expression and protein secretion in a concentration-dependent manner. Proportional induction of PAI-1 promoter activity was observed through increasing amounts of PPARα DNA in HUVECs through a transient gene transfection assay, although the mRNA expression of the 3 subtypes of PPAR with their activators were not changed compared with controls.Conclusions HUVECs express PPARs. PPARs activators may increase PAI-1 expression in endothelial cells (EC). Although PPARs expression was not enhanced after being stimulated by their activators in EC, the functionally active PPARα is probably involved in regulating PAI-1 expression in EC.

Relationship of plasminogen activator inhibitor 1 gene 4G/5G polymorphisms to hypertension in Korean women

Background Hypertension （HTN） is a major determinant of various cardiovascular events. Plasma levels of plasminogen activator inhibitor 1 （PAl-l） modulate this risk. A deletion/insertion polymorphism within the PAl-1 loci （4G/4G, 4G/5G, 5G/5G） affects the expression of this gene. The present study investigated the association between PAl-1 loci polymorphisms and HTN in Korean women. Methods Korean women （n=1312） were enrolled in this study to evaluate the association between PAl-1 4G/5G gene polymorphisms and HTN as well as other metabolic risk factors. PAl-1 loci polymorphisms were investigated using polymerase chain reaction amplification and single-strand conformation polymorphism analysis. Results The three genotype groups differed with respect to systolic blood pressure （P=0.043）, and diastolic blood pressure （P=0.009） but not with respect to age, body mass index, total cholesterol, low or high density lipoprotein cholesterol, triglycerides, or fasting blood glucose. Carriers of the PAl-1 4G allele had more hypertension significantly （PAl-1 4G/5G vs. PAl-1 5G/5G, P=0.032; PAl-1 4G/4G vs. PAl-1 5G/5G, P=0.034）. When stratified according to PAl-1 4G/5G polymorphism, there was no significant difference in all metabolic parameters among PAl-1 genotype groups in patients with HTN as well as subjects with normal blood pressure. The estimated odds ratio of the 4G/4G genotype and 4G/5G for HTN was 1.7 （P=0.005）, and 1.6 （P=0.015）, respectively. Conclusion These findings might indicate that PAl-1 loci polymorphisms independently contribute to HTN and that gene-environmental interaction may be not associated in Korean women.

Plasminogen activator inhibitor-1 4G/5G gene polymorphism in patients with myocardial or cerebrovascular infarction in Tianjin, China

Objective To investigate the association between the plasminogen activator inhibitor-1 (PAI-1) 4G/ 5G gene polymorphism and the occurrence of myocardial and cerebrovascular infarctions in individuals from Tianjin, China.Methods The PAI-1 genotype was determined using allele-specific polymerase chain reaction (AS-PCR) in 56 myocardial infarction (Ml) patients, 54 cerebrovascular infarction (Cl) patients and 83 unrelated healthy controls. All subjects' clinical features and plasma PAI-1 activity levels were determined.Results The PAI-1 genotype distribution frequency of the single guanine deletion/insertion 4G/5G polymorphism (located -675 bp upstream from the start of transcription) significantly differed between the patients and healthy controls. In the Ml group, the4G/4G-genotype frequency was increased, but the 4G/5G-genotype is decreased when compared to the control group. In the Cl group, both the 4G/ 4G- and 4G/5G -genotypes occured at a lower frequency than those in the control group (P<0. 001) . The plasma PAI-1 activity level in the Ml group was lowered as the presence of the 4G allele decreases. In the Cl group, the frequency of 5G/5G was much higher than that of the control group (P<0. 001). The plasma PAI-1 activity level in the Cl group was elevated as the presence of the 5G allele increased. Furthermore, positive correlation between triglyceride, glucose levels and PAI-1 activity were found in all three groups (P<0. 001).Conclusions The PAI-1 4G/5G gene polymorphism is associated with a higher risk of Ml and Cl in individuals in Tianjin, China. The deletion/insertion polymorphism is probably an important hereditary risk factor for heart diseases. Moreover, triglyceride and glucose levels of plasma have functional importance in regulating PAI-1 activity.

Gene expression of fibrinolytic factors urokinase plasminogen activator and plasminogen activator inhibitor-1 in rabbit temporo-mandibular joint cartilage with disc displacement

Background The urokinase plasminogen activator system is believed to play an important role in degradation of the extracellular matrix associated with cartilage and bone destruction; however its precise roles in temporomandibular disorders have not yet been clarified. The aims of this study were to investigate the gene expression of fibrinolytic factors urokinase plasminogen activator (uPA) and plasminogen activator inhibitor-1 (PAI-1) in the articular cartilage of rabbit temporomandibular joint (TMJ) with disc displacement (DD) and to probe the relationship between fibrinolytic activity and cartilage remodeling. Methods Disc displacement of right joints was performed in 36 of 78 rabbits under investigation. The animals were sacrificed at 4 days and 1, 2, 4, 8 and 12 weeks after surgery, respectively. The right joints of these animals were harvested and processed for the examination of mRNA expression of uPA and PAI-1 in articular cartilage using in situ hybridization techniques. Results The expression of uPA and PAI-1 was co-expressed weakly in the chondrocytes from transitive zone to hypertrophic zone and mineralized zone, while no hybridizing signals were shown in proliferative zone and superficial zone in control rabbits. The most striking was the up-regulation of uPA and PAI-1 mRNA in 4-day rabbits postoperatively at the onset of cartilage degeneration. The strongest hybridizing signals for uPA and PAI-1 were seen in 2-week rabbits postoperatively. After 2 weeks, the expression of uPA and PAI-1 began to decrease and reached nearly normal level at 12 weeks. Conclusions The expression of the uPA/PAI-1 system coincides with the pathological changes in condylar cartilage after DD. The uPA/PAI-1 system may be one of the essential mediators in articular cartilage remodeling.

Alterations in fibrinolytic system proteins PAI-1,MMP-3,MMP-8,TIMP-1 and TIMP-2 in post-cholecystectomy bile duct injury

Introduction: In bile duct injuries (BDI), cholestasis and cholangitis can alter the fibrinolytic system by promoting an increase of extracellular matrix depositions which favor an imbalance between metalloproteinases (MMPs) and their tissue inhibitors (TIMPs). Materials and Methods: Levels of PAI-1, MMP-3, MMP-8, TIMP-1 and TIMP-2 in 35 patients with post-cholecystectomy BDI by complete biliary obstruction were measured and compared to a healthy control group. Sirius red staining and immune staining for MMP-3 and MMP-8 were also undertaken in liver biopsies. Results: Levels of PAI-1, TIMP-1, TIMP-2 and MMP-8 were higher in BDI than healthy controls: 15 ± 2 ng/mL vs 7.1 ± 2 ng/mL (p 0.024);539 ± 64 ng/mL vs 256 ± 13 ng/mL (p p p 2 vs. 22865.7 ± 3865 μm2 in healthy controls (p 2 vs. 30744.2 ± 5810.2 μm2 (p 2 vs. 116337.9 ± 24803.3 μm2 (p 0.55). These results suggest an imbalance between fibrogenic/fibrinolytic protein levels. Interestingly, expression of the fibrinolytic protein MMP-8 was increased in serum and liver biopsies in BDI. Conclusion: We found an imbalance of profibrogenic molecules which promote extracellular matrix deposition. The over-expression of fibrinolytic proteins such as MMP-8 could limit liver fibrosis, preventing hepatic dysfunction in post-cholecystectomy BDI.

The Physiopathological Crossroads of Aging

Stress, inflammation and Plasminogen activator inhibitor 1 (PAI-1) are key mechanisms throughout the development of aging, constituting a crossroad in the most frequent pathologies that accompany it. Among metabolic processes, obesity, metabolic syndrome and type 2 diabetes mellitus are included and Alzheimer’s disease among the neurodegenerative processes. Stress is a mechanism of defense of the organism against exogenous and endogenous actions called stressors. In the case of low intensity stimuli, the organism responds with actions aimed at a physiological adaptation (Homeostasis). On the other hand, when a high intensity (experimental level) or chronic stimulus (oxidative stress) is repeated, structural and functional changes are observed in different organs with activation of the hypothalamus-pituitary-adrenal axis, the renin angiotensin system and the sympathetic nervous system, stimulating the production of hormones that release cytokines with proin-flammatory/antiinflammatory properties that play an important role in the previously mentioned pathologies, as well as a marked increase in PAI-1, a gene regulated by stress and by cytokines, with manifest action at the origin of thromboembolic disease, so frequent in aging. The objective of this review is to highlight the importance of the binomial stress and PAI-1 in aging and in the pathologies that accompany it. Because PAI-1 is part of the pathology and complications in aging, some authors suggest the study of PAI-1 inhibitors to achieve its physiological levels, as part of the treatment of these diseases.

Effects of Modified Qing'e Pill(加味青娥丸) on Expression of Adiponectin,Bone Morphogenetic Protein 2 and Coagulation-Related Factors in Patients with Nontraumatic Osteonecrosis of Femoral Head

Objective： To observe the regulation of Chinese herbal medicine, Modified Qing＇e Pill（加味青娥丸, MQEP）, on the expression of adiponectin, bone morphogenetic protein 2（BMP2）, osteoprotegerin（OPG） and other potentially relevant risk factors in patients with nontraumatic osteonecrosis of the femoral head（ONFH）. Methods： A total of 96 patients with nontraumatic ONFH were unequal randomly divided into treatment group（60 cases） and control group（36 cases）. The treatment group were treated with MQEP while the control group were treated with simulated pills. Both groups were given caltrate D. Six months were taken as a treatment course. Patients were followed up every 2 months. The levels of plasma adiponectin, BMP2, OPG, von Willebrand factor（vWF）, von Willebrand factor cleaving protease（vWF-cp）, plasminogen activator inhibitor 1（PAI-1）, tissue plasminogen activator（tPA）, C-reactive protein（CRP）, blood rheology, bone mineral density（BMD） of the femoral head and Harris Hip Score were measured before and after treatment. Results： After 6 months of treatment, compared with the control group, patients in the treatment group had significantly higher adiponectin and BMP2 levels（P〈0.01 and P=0.013, respectively）, lower vWF, PAI-1 and CRP levels（P=0.019, P〈0.01 and P〈0.01, respectively）, and lower blood rheology parameters. BMD of the femoral neck, triangle area and Harris Hip Score in the treatment group were significantly higher than those in the control group. Moreover, plasma adiponectin showed a positive association with BMP2（r=0.231, P=0.003） and a negative association with PAI-1（r=–0.159, P〈0.05）. Conclusions： MQEP may play a protective role against nontraumatic ONFH by increasing the expression of adiponectin, regulating bone metabolism and improving the hypercoagulation state, which may provide an experimental base for its clinical effects.

Clinical and genetic risk factors for venous thromboembolismin Chinese population

Venous thromboembolism(VTE),including deep vein thrombosis and pulmonary embolism,carries significant mortality and morbidity.The most important and effective way to reduce VTE incidence is to identify the patients at risk and give necessary prevention.VTE is a multifactorial and complicated disorder.Major risk factors for VTE include surgery and trauma,acute medical illness,active cancer and pregnancy.Genetic factors increase susceptibility to the disease and are useful in predicting the development of VTE.Gene-gene and gene-environment interactions alter and magnify the clinical picture in this disorder.This brief review summarizes some selected clinical and genetic risk factors for VTE based on the current research in China.

Effects of pioglitazone on arteriosclerotic-related factors and short term prognosis in patients with coronary disease and diabetes

Background Thiazolidinediones （TZDs） not only improve insulin resistance, lowering blood sugar, also has anti-atherosclerotic effect. However, whether the protective effect on cardiovascular pioglitazone is still controversial. Methods Totally 98 patients with coronary disease and diabetes mellitus were randomly divided into pioglitazone group （n = 48） receiving conventional therapy and pioglitazone （15 mg/day）, and control group （n = 50） merely receiving conventional therapy. The patients were followed up for 12 months. The plasma level of Plasminogen activator Inhibitor 1 （PAI-1） and P-selectin were detected at baseline and after treatment for 12 months by ELISA, and major adverse cardiac events （MACE） were studied. Results Pioglitazone therapy for 12 months was associated with a significant decrease of PAI-1 [（7.9 ± 1.4 vs 4.2 ± 0.5）ng/mL, P 〈 0.05] and P-selectin [（16.6 ± 6.8 vs 12.4 ± 3.6）ng/mL, P 〈 0.05], MACE was significantly lower in the pioglitazone group than in the control group [acute coronary syndrome （ACS）： 32.0% vs 10.4%, P 〈 0.05; target vessel revascularization： 22.0% vs 6.3%, P 〈 0.05 ]. Conclusions Pioglitazone can effectively reduce the plasma level of PAI-1, P-selectin and the occurrence of MACE in patients with coronary heart disease and diabetes mellitus.