Hypoxia, as an important hallmark of the tumor microenvironment, is a major cause of oxidative stress and plays a central role in various malignant tumors, including glioblastoma. Elevated reactive oxygen species (ROS) in a hypoxic microenvironment promote glioblastoma progression; however, the underlying mechanism has not been clarified. Herein, we found that hypoxia promoted ROS production, and the proliferation, migration, and invasion of glioblastoma cells, while this promotion was restrained by ROS scavengers N-acetyl-L-cysteine (NAC) and diphenyleneiodonium chloride (DPI). Hypoxia-induced ROS activated hypoxia-inducible factor-1α (HIF-1α) signaling, which enhanced cell migration and invasion by epithelial-mesenchymal transition (EMT). Furthermore, the induction of serine protease inhibitor family E member 1 (SERPINE1) was ROS-dependent under hypoxia, and HIF-1α mediated SERPINE1 increase induced by ROS via binding to the SERPINE1 promoter region, thereby facilitating glioblastoma migration and invasion. Taken together, our data revealed that hypoxia-induced ROS reinforce the hypoxic adaptation of glioblastoma by driving the HIF-1α-SERPINE1 signaling pathway, and that targeting ROS may be a promising therapeutic strategy for glioblastoma.
Humans ; Cell Hypoxia ; Cell Line, Tumor ; Glioblastoma/pathology* ; Hypoxia-Inducible Factor 1, alpha Subunit/metabolism* ; Plasminogen Activator Inhibitor 1/metabolism* ; Reactive Oxygen Species/metabolism* ; Signal Transduction ; Tumor Microenvironment ; Brain Neoplasms/pathology*

OBJECTIVETo explore the mechanism of the protective effects of Panax notoginseng saponins (PNS) on kidney in diabetic rats.

METHODSDiabetic rat model was obtained by intravenous injection of alloxan, and the rats were divided into model, PNS-100 mg/(kg day) and PNS-200 mg/(kg day) groups, 10 each. Another 10 rats injected with saline were served as control. Periodic acid-Schiff staining and immunological histological chemistry were used to observe histomorphology and tissue expression of bone morphogenetic protein-7 (BMP-7). Silent information regulator 1 (SIRT1) was silenced in rat mesangial cells by RNA interference. The mRNA expressions of SIRT-1, monocyte chemoattractant protein-1 (MCP-1), transforming growth factor β1 (TGF-β1) and plasminogen activator inhibitor-1 (PAI-1) were analyzed by reverse transcription polymerase chain reaction. The protein expressions of SIRT1 and the acetylation of nuclear factor κB (NF-κB) P65 were determined by western blotting. The concentration of MCP-1, TGF-β1 and malondialdehyde (MDA) in culture supernatant were detected by enzyme-linked immuno sorbent assay. The activity of superoxide dismutase (SOD) was detected by the classical method of nitrogen and blue four.

RESULTSIn diabetic model rats, PNS could not only reduce blood glucose and lipid (P<0.01), but also increase protein level of BMP-7 and inhibit PAI-1 expression for suppressing fibrosis of the kidney. In rat mesangial cells, PNS could up-regulate the expression of SIRT1 (P<0.01) and in turn suppress the transcription of TGF-β1 (P<0.05) and MCP-1 (P<0.05). PNS could also reverse the increased acetylation of NF-κB p65 by high glucose. In addition, redox regulation factor MDA was down-regulated (P<0.05) and SOD was up-regulated (P<0.01), which were both induced by SIRT1 up-regulation.

CONCLUSIONSPNS could protect kidney from diabetes with the possible mechanism of up-regulating SIRT1, therefore inhibiting inflammation through decreasing the induction of inflammatory cytokines and TGF-β1, as well as activating antioxidant proteins.

Acetylation ; drug effects ; Animals ; Antioxidants ; metabolism ; Blood Glucose ; metabolism ; Bone Morphogenetic Protein 7 ; metabolism ; Chemokine CCL2 ; metabolism ; Diabetes Mellitus, Experimental ; blood ; drug therapy ; genetics ; physiopathology ; Gene Knockdown Techniques ; Immunohistochemistry ; Kidney ; drug effects ; pathology ; Kidney Function Tests ; Lipids ; blood ; Male ; Malondialdehyde ; metabolism ; Mesangial Cells ; drug effects ; metabolism ; Oxidative Stress ; drug effects ; Panax notoginseng ; chemistry ; Plasminogen Activator Inhibitor 1 ; genetics ; metabolism ; Protective Agents ; pharmacology ; therapeutic use ; Rats, Sprague-Dawley ; Saponins ; pharmacology ; therapeutic use ; Sirtuin 1 ; genetics ; Superoxide Dismutase ; metabolism ; Transcription Factor RelA ; metabolism ; Transcription, Genetic ; drug effects ; Transforming Growth Factor beta1 ; metabolism ; Up-Regulation ; drug effects

OBJECTIVETo evaluate the effect of sodium tanshinone IIA sulfonate (STS) in preventing postoperative peritoneal adhesions in rats and explore the mechanisms.

METHODSSixty SD rats were randomized into 4 equal groups, including a blank control group, adhesion model group, and high-, moderate-, and low-dose STS-treated groups, and were subjected to injuries of the parietal peritoneum and cecum to induce peritoneal adhesions, followed by intraperitoneal administration of saline and STS at the doses of 20, 10, and 5 mg/kg for 7 consecutive days, respectively. Another 15 untreated rats served as the blank control group. The adhesion scores in each group were recorded after the treatments; the activity of tissue-type plasminogen activator (tPA) in peritoneal lavage fluid was measured, tPA/PAI-1 protein ratio in the peritoneal tissue was determined by ELISA, and the expressions of TGF-β1 and collagen I were detected by immunohistochemistry. The anastomotic healing model was used to assess the impact of STS on wound healing.

RESULTSIntraperitoneal administration of STS effectively prevented peritoneal adhesion without affecting anastomotic healing in the rats. Compared with the adhesion model group, the STS-treated groups showed increased peritoneal lavage fluid tPA activity and tPA/PAI-1 ratio in the ischemic tissues with lowered TGF-β1 and collagen I expressions in the ischemic tissues.

CONCLUSIONSIntraperitoneal administration of STS can prevent peritoneal adhesion and enhance local fibrinolysis in rats, and these effects may be mediated by TGF-β signaling pathway.

Animals ; Cecum ; injuries ; pathology ; Collagen Type I ; metabolism ; Fibrinolysis ; Injections, Intraperitoneal ; Peritoneum ; injuries ; pathology ; Phenanthrenes ; pharmacology ; Plasminogen Activator Inhibitor 1 ; metabolism ; Postoperative Complications ; prevention & control ; Rats ; Rats, Sprague-Dawley ; Tissue Adhesions ; prevention & control ; Tissue Plasminogen Activator ; metabolism ; Transforming Growth Factor beta1 ; metabolism ; Wound Healing

PURPOSE: Dysregulation of adipokines caused by excess adipose tissue has been implicated in the development of obesity-related metabolic diseases. This study evaluated the effects of mugwort (Artemisia princeps Pampanini) ethanol extract on lipid metabolic changes, insulin resistance, adipokine balance, and body fat reduction in obese rats. METHODS: Male Sprague-Dawley rats were fed either a control diet (NC), high-fat diet (HF, 40% kcal from fat), or high-fat diet with 1% mugwort extract (HFM) for 6 weeks. RESULTS: Epididymal and retroperitoneal fat mass increased in the HF group compared with the NC group, and epididymal fat mass was reduced in the HFM group (p < 0.05). No difference was observed in serum levels of total cholesterol (TC) and high-density lipoprotein cholesterol (HDL-C) among the groups. However, triglyceride (TG), TG/HDL-C ratio, and TC/HDL-C ratio increased in the HF group and significantly decreased in the HFM group. TG and TC levels in the liver were significantly higher in the HF group, whereas these levels were significantly reduced in the HFM group. HF rats had lower insulin sensitivity as indicated by increased homeostasis model assessment of the insulin resistance (HOMA-IR) value. HOMA-IR values significantly decreased in the HFM group. Adiponectin levels were higher in NC rats, and their leptin and PAI-1 levels were lower. Relative balance of adipokines was reversed in the HF group, with lower adiponectin levels but higher leptin and PAI-1 levels. In contrast, the HFM group maintained balance of adiponectin/leptin and adiponectin/PAI-1 levels similar to NC by reducing leptin and PAI-1 levels. CONCLUSION: Overall data indicated that mugwort extract can be effective in alleviating metabolic dislipidemia, insulin resistance, and adipokine dysregulation induced by a high-fat diet.
Adipokines* ; Adiponectin ; Adipose Tissue ; Animals ; Artemisia* ; Cholesterol ; Diet ; Diet, High-Fat ; Ethanol ; Homeostasis ; Humans ; Insulin Resistance ; Intra-Abdominal Fat ; Leptin ; Lipid Metabolism ; Lipoproteins ; Liver ; Male ; Metabolic Diseases ; Plasminogen Activator Inhibitor 1 ; Rats* ; Rats, Sprague-Dawley ; Triglycerides

OBJECTIVEThe exact mechanisms of defect closure in patients with perimembranous ventricular septal defect (PMVSD) remain unknown. We hypothesized that the expression of urokinase type plasminogen activator (uPA) and plasminogen activator inhibitor-1 (PAI-1) may mediate extracellular matrix (ECM) remodeling in aneurysms.

METHODSeven normal heart tricuspid septal leaflet and 33 aneurysms were collected in Shanghai Renji Hospital and Shanghai Children's Medical Center from January 2008 to June 2010. Immunohistochemical expression of uPA and PAI-1 in 4 normal heart valvular tissues and 15 aneurysms was detected with immunohistochemical methods. The expression of uPA and PAI-1 mRNA in 3 normal heart valvular tissues and 7 aneurysms was studied by real time fluorescent PCR; the protein expression of uPA and PAI-1 in 4 normal heart valvular tissues and 11 aneurysms was tested with Western blotting.

RESULTThe surface of the aneurysms were completely covered by endothelial cells. Two types of granulation tissue, myxoid and fibrous, were associated with the aneurismal formation. uPA were recognized predominantly in valvar interstitial cells (VICs) which located mainly in regions adjacent to the endothelium and smooth muscle cells of blood vessels. PAI-1 was found in both VICs which located mainly in granulation tissue and endothelial cells. Nine aneurysms expressed a higher uPA activity than 4 normal valvular tissues ((74.6±11.8)% vs. (49.5±7.4)%; t = 3.87, P = 0.003) and six aneurysms expressed a low uPA activity ((10.3±3.1)% vs. (49.5±7.4)%; t=11.78, P=0.000) and a high PAI-1 activity ((55.2±1.7)% vs. (50.8±3.8)%; t=2.55, P=0.034) using immunohistochemical methods. uPA / PAI-1 ratio of protein expression tested by Western blot was 0.88±0.22 in four normal heart vavular tissues; five aneurysms expressed high uPA activity and low PAI-1 activity and uPA/PAI-1 ratio was 4.26±2.04; while the other 6 cases expressed low uPA activity and high PAI-1 activity and uPA/PAI-1 ratio was 0.30±0.07; the difference among the three groups was statistically significant (P<0.05). The rate of uPA/PAI-1 in relative copy of mRNA expression among normal heart valvular tissue, high uPA expressed aneurysms and low uPA expressed aneurysms are also significantly different (2.14±0.17 vs. 0.45±0.04; 2.14±0.17 vs. 4.38±1.41, P<0.05) respectively.

CONCLUSIONThe expression of uPA and PAI-1 in VICs suggests that interactions among these molecules contribute to the aneurysm formation and development. This provides a potential mechanism for defect closure in patients with PMVSD.

Aneurysm ; pathology ; Blotting, Western ; China ; Endothelial Cells ; cytology ; Extracellular Matrix ; metabolism ; Granulation Tissue ; pathology ; Heart Septal Defects, Ventricular ; pathology ; Humans ; Immunohistochemistry ; Plasminogen Activator Inhibitor 1 ; metabolism ; RNA, Messenger ; Urokinase-Type Plasminogen Activator ; metabolism

OBJECTIVETo study the expression of transforming growth factor-β1 (TGF-β1) and plasminogen activator inhibitor-1 (PAI-1) and its significance in premature infants with bronchopulmonary dysplasia (BPD).

METHODSA retrospective analysis was performed on the clinical data of 96 very low birth weight infants (gestational age of ≤ 32 weeks) who survived for more than 28 days and were admitted to the Neonatal Intensive Care Unit between January 2010 and December 2012. These subjects were divided into BPD group (n=21) and non-BPD group (n=75). The expression of TGF-β1 and PAI-1 in blood was measured by ELISA.

RESULTSThe levels of TGF-β1 and PAI-1 in the BPD group increased gradually from the 7th day to the 14th day and then to the 21st day after birth, and were significantly higher than in the non-BPD group at all time points (P<0.01). The TGF-β1 and PAI-1 levels in the non-BPD group on the 7th, 14th, and 21st days after birth were not significantly different from each other (P>0.05).

CONCLUSIONSThe expression of TGF-β1 and PAI-1 in blood is elevated in premature infants with BPD, which may be associated with the development of BPD.

Bronchopulmonary Dysplasia ; etiology ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Infant, Newborn ; Infant, Premature ; Male ; Plasminogen Activator Inhibitor 1 ; blood ; Retrospective Studies ; Transforming Growth Factor beta1 ; blood

BACKGROUND: Gestational diabetes mellitus (GDM) is a hyperglycemic condition caused by increased insulin resistance and impaired insulin secretion during pregnancy. It is known to be temporary, but it can cause perinatal complications in the mother and baby. Additionally, it may progress to type 2 diabetes mellitus (T2DM). In the present study, we evaluated the risk factors for complications and progression to T2DM in patients with GDM. METHODS: The study included 130 pregnant women who were diagnosed with GDM at gestational weeks 24-28 in 2011. Body mass index and the levels of glucose, total cholesterol, lipoproteins, and coagulation factors (von Willebrand factor and plasminogen activator inhibitor-1) were assessed in all patients. RESULTS: The level of high-density lipoprotein (HDL) was significantly lower and the triglyceride/HDL ratio and coagulation factor levels were significantly higher in the group of patients with perinatal complications compared to those in the group of patients without complications. After delivery, the level of HDL was lower and the value of homeostasis model assessment of insulin resistance (HOMA-IR) was higher in women with impaired glucose metabolism compared to those in women with normal glucose metabolism. In logistic regression analysis, perinatal complications were independently associated with HDL and PAI-1 levels (OR = 0.929 and 1.101, respectively). CONCLUSION: The findings of our study show that the levels of HDL and coagulation factors are notable risk factors of perinatal complications. Additionally, we showed that lower HDL level may influence the progression to T2DM. Large-scale population studies are needed to verify our findings.
Blood Coagulation Factors ; Body Mass Index ; Cholesterol ; Diabetes Mellitus* ; Diabetes Mellitus, Type 2 ; Diabetes, Gestational* ; Female ; Glucose ; Homeostasis ; Humans ; Insulin ; Insulin Resistance ; Lipoproteins ; Lipoproteins, HDL ; Logistic Models ; Metabolism ; Mothers ; Plasminogen Activator Inhibitor 1 ; Plasminogen Activators ; Postpartum Period* ; Pregnancy ; Pregnant Women ; Risk Factors* ; von Willebrand Factor

BACKGROUNDStent thrombosis is one of severe complications after sirolimus-eluting stent implantation. Rapamycin (sirolimus) promotes arterial thrombosis in in vivo studies. However, the underlying molecular and transcriptional mechanisms of this adverse effect have not been thoroughly investigated. This study was designed to examine the effects of rapamycin on the expression of the gene, Kruppel-like factor 2 (KLF2), and its transcriptional targets in mice.

METHODSMice were randomly divided into four groups: the control group (intraperitoneal injection with 2.5% of dimethyl sulfoxide (DMSO) only), rapamycin group (intraperitoneal injection with 2 mg/kg of rapamycin only), Ad-LacZ + rapamycin group (carotid arterial incubation with Ad-LacZ plus intraperitoneal injection with 2 mg/kg of rapamycin 10 days later), and Ad-KLF2 + rapamycin group (carotid arterial incubation with Ad-KLF2 plus intraperitoneal injection with 2 mg/kg rapamycin 10 days later). The carotid arterial thrombosis formation was induced by FeCl3 and the time of arterial thrombosis was determined. Finally, the RNA and protein of carotid arteries were extracted for KLF2, tissue factor (TF), plasminogen activator inhibitor-1 (PAI-1), endothelial nitric oxide synthase (eNOS), thrombomodulin (TM) mRNA and protein analysis.

RESULTSCompared with controls, treatment with rapamycin inhibited KLF2, eNOS and TM mRNA and protein expression, and enhanced TF and PAI-1 mRNA and protein expression, and shortened time to thrombotic occlusion from (1282 ± 347) seconds to (715 ± 120) seconds (P < 0.01) in vivo. Overexpression of KLF2 strongly reversed rapamycin-induced effects on KLF2, eNOS, TM, TF and PAI-1 expression. KLF2 overexpression increased the time to thrombotic occlusion to control levels in vivo.

CONCLUSIONSRapamycin induced an inhibition of KLF2 expression and an imbalance of anti- and pro-thrombotic gene expression, which promoted arterial thrombosis in vivo. Overexpression of KLF2 increased KLF2 expression and reversed time to thrombosis in vivo.

Animals ; Carotid Arteries ; metabolism ; Drug-Eluting Stents ; adverse effects ; Kruppel-Like Transcription Factors ; analysis ; genetics ; physiology ; Mice ; Mice, Inbred C57BL ; Nitric Oxide Synthase Type III ; physiology ; Plasminogen Activator Inhibitor 1 ; physiology ; Sirolimus ; pharmacology ; Thrombomodulin ; physiology ; Thrombosis ; chemically induced

In order to discover the mechanism of Xuebijing oral effervescent tablet (XBJOET) to treat infectious diseases, the effect of XBJOET on endotoxin induced rabbit fever and disseminated intravascular coagulation (DIC) was investigated. Auricle microcirculation in rabbit was detected by laser speckle blood perfusion imager system; coagulation function was measured by coagulation analyzer, fibrinolytic system was quantified by Elisa assay and micro thrombosis in tissues was observed with HE staining under light microscope. The results demonstrated that the body temperature of rabbit decreased significantly at 1-3 h after administration with 4.8, 2.4 and 1.2 g x kg(-1) XBJOET to endotoxin induced DIC rabbit model, the auricle microcirculation blood flow in model group (54.45 +/- 14.53) PU was lower than that in control group (77.18 +/- 12.32) PU. The auricle microcirculation blood flow increased markedly and there was significant difference between model group and 1.2 g x kg(-1) XBJOET group. There was significant difference between model group and control group in the content of PAI1 and FIB. The PAI1 levels in model and control groups were (30.48 +/- 2.46) ng x mL(-1) and (20.93 +/- 3.25) ng x mL(-1), respectively. The FIB levels in model and control group were (3.34 +/- 1.09) g x L(-1) and (4.84 +/- 1.10) g x L(-1), respectively. The content of PAI1 in rabbit plasma decreased notably, there were significant differences between model group and 4.8, 2.4 g x kg(-1) XBJOET groups. On the contrary the content of FIB increased. XBJOET possessed pharmacological activities of curing infectious fever and DIC, the mechanism of which is related to amelioration of microcirculation disturbance, inhibition of fibrinolytic system activation and coagulation and micro thrombosis elimination.
Administration, Oral ; Animals ; Blood Coagulation ; drug effects ; Body Temperature ; drug effects ; Disseminated Intravascular Coagulation ; blood ; chemically induced ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Ear Auricle ; blood supply ; Endotoxins ; Female ; Fever ; chemically induced ; drug therapy ; physiopathology ; Fibrinogen ; metabolism ; Male ; Microcirculation ; Partial Thromboplastin Time ; Plasminogen Activator Inhibitor 1 ; blood ; Prothrombin Time ; Rabbits ; Tablets ; Thrombosis ; pathology

OBJECTIVETo assess the influence of laparoscopic colorectal cancer resection on the peritoneal microstructure injury and expression of t-PA/PAI-1 molecules.

METHODSA total of 50 patients with colorectal cancer were prospectively enrolled between June 2011 and February 2012 in the Shanxi Provincial Hospital and were assigned into laparoscopic group (LO, n=27) and conventional laparotomy group (CO, n=23) based on patients expectancy and surgeon decision. Optical microscope and scanning electron microscope were employed for comparison of the postoperative peritoneal injury between LO and CO. Before and after surgery, t-PA and PAI-1 of peritoneal tissue were determined by ELISA in both groups.

RESULTSOptical microscope and scanning electronic microscopy scan indicated less serosal injury in LO group than that in CO group with regard to serosa integrity, continuity of covering adipocytes and mesothelial cells, and the aggregation level of inflammatory cells (P<0.01). The injury score was 38.22 in CO in and 14.67 in LO and the difference was statistically significant (P<0.01). No significant differences were found between LO and CO in terms of postoperative t-PA in the omentum, t-PA and PAI-1 in the intestinal serosa tissue (P>0.05), however PAI-1 in the omentum was significantly lower in LO group compared to CO group (P<0.05).

CONCLUSIONLaparoscopic radical resection for colorectal cancer causes less peritoneal structural injury and less influence on the fibrinolytic capacity, which may contribute to less postoperative adhesion.

Adolescent ; Adult ; Aged ; Colorectal Neoplasms ; metabolism ; surgery ; Colorectal Surgery ; adverse effects ; methods ; Female ; Humans ; Laparoscopy ; adverse effects ; Male ; Middle Aged ; Peritoneum ; metabolism ; pathology ; Plasminogen Activator Inhibitor 1 ; metabolism ; Prospective Studies ; Tissue Plasminogen Activator ; metabolism ; Young Adult