Animals ; Ethanol ; pharmacology ; Female ; Fetal Heart ; drug effects ; growth & development ; Male ; Zebrafish ; embryology

OBJECTIVETo improve the biological properties of decellularized aortic valves by polyethylene glycol (PEG)-mediated covalent incorporation of vascular endothelial growth factor (VEGF).

METHODSPEG crosslinking of decellularized aortic valves were completed via a Michael-type addition reaction, followed by covalent incorporation of VEGF through another Michael-type addition reaction between the unsaturated propylene acyl of PEG and the thiol groups on cysteine residues of VEGF. The effect of VEGF incorporation was evaluated by enzyme-linked immunosorbent assay (ELISA) and immune fluorescence assay. The endothelial progenitor cells (EPCs) were seeded on decellularized aortic valves with or without these modifications, and after 10 days of culture, the valves were examined for DNA content and by hematoxylin-eosin staining and scanning electron microscopy.

RESULTSImmune fluorescence and ELISA showed that the maximal VEGF incorporation on the decellularized aortic valve reached 908.94∓0.27 pg. Compared with the unmodified valves and the valves with PEG crosslinking, decellularized aortic valves with covalent incorporation of VEGF significantly promoted the adhesion and proliferation of EPCs, which formed a confluent cell monolayer on the valve surface.

CONCLUSIONSPEG-mediated covalent incorporation of VEGF in the decellularized aortic valves improves the adhesion and proliferation of the seeded EPCs to facilitate the construction of tissue-engineered heart valves.

Animals ; Aortic Valve ; drug effects ; Cell Adhesion ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Endothelial Cells ; cytology ; drug effects ; Heart Valve Prosthesis ; Polyethylene Glycols ; pharmacology ; Stem Cells ; cytology ; drug effects ; Swine ; Tissue Engineering ; Vascular Endothelial Growth Factor A ; pharmacology

OBJECTIVETo observe the effect of sodium tanshinone II (A) sulfonate (STS) on Ang II -induced atrial fibroblast collagen synthesis and TGF-beta1 activation.

METHODAtrial fibroblasts of neonatal rats were cultured to determine the content of collagen protein. The original synthesis rate determined by the [3H]-proline incorporation method was taken as the index for myocardial fibrosis. The content of active TGF-beta1 and total TGF-beta1 in cell culture supernatants were tested and cultured by ELISA. The expression of thrombospondin-1 (TSP-1) was assessed by using Western blot.

RESULTAng II could significantly increase the content of atrial fibroblast collagen and the collagen synthesis rate, the TSP-1 expression and the concentration of active TGF-beta1, without any obvious change in total TGF-beta1. After the STS treatment, all of the indexes, apart from total TGF-beta1, were obviously down-regulated.

CONCLUSIONSTS could decrease the secretion of Ang II -induced atrial fibroblast collagen and the synthesis rate. Its mechanism is related to the inhibition of TSP-1/TGF-beta1 pathway.

Angiotensin II ; pharmacology ; Animals ; Collagen ; biosynthesis ; Fibroblasts ; cytology ; drug effects ; metabolism ; Gene Expression Regulation ; drug effects ; Heart Atria ; cytology ; Phenanthrenes ; pharmacology ; Rats ; Rats, Wistar ; Signal Transduction ; drug effects ; Thrombospondin 1 ; metabolism ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVETo investigate the effect of atorvastatin on cardiac remodeling and function after acute myocardial infarction (AMI) in rats and whether this effect is mediated by transforming growth factor-β1 (TGF-β1) signaling pathway.

METHODSAMI was induced by left coronary artery ligation in 64 male Sprague-Dawley rats, and 45 surviving rats were randomized into control group (n=15), low-dose atorvastatin group (10 mg/kg, n=15) and high-dose atorvastatin group (20 mg/kg, n=15). Similar surgical procedure was performed in sham-operated rats (n=15) without coronary ligation. Atorvastatin was given daily by gavage from the first day after AMI. Eight weeks later, the cardiac function, left ventricular weight/body mass index (LVMI), collagen volume fraction (CVF), and the expressions of TGF-β1 and Smad2 were compared between the groups.

RESULTSAMI caused significantly reduced cardiac function, increased LVMI and CVF, and upregulated expressions of TGF-β1 and Smad2 mRNA and proteins in the control group (P<0.05). The cardiac function, LVMI, and CVF were improved by atorvastatin, which also down-regulated the expressions of TGF-β1 and Smad2 (P<0.05), and the effects were more prominent in high-dose atorvastatin group (P<0.05).

CONCLUSIONAtorvastatin can dose-dependently improve cardiac remodeling and function after AMI in rats, which is mediated by regulating the activity of TGF-β1/Smad2 signaling pathway.

Animals ; Atorvastatin Calcium ; Heart ; drug effects ; physiopathology ; Heptanoic Acids ; pharmacology ; Male ; Myocardial Infarction ; physiopathology ; Pyrroles ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; Smad2 Protein ; metabolism ; Transforming Growth Factor beta1 ; metabolism ; Ventricular Remodeling ; drug effects

OBJECTIVETo investigate the effects of metoprolol on electrophysiology of ischemic and anoxic myocardium in diabetic rats.

METHODSForty Sprague-Dawley (SD) rats were divided into 4 groups: diabetes group; diabetes and ablation of left sympathetic nerve group; diabetes and metoprolol group and sham group. The diabetes model was induced by intraperitoneal injection of streptozotocin (STZ, 60 mg/kg). The ventricular diastolic effective threshold (DET), effective refractive period (ERP), and Ventricular fibrillation threshold (VFT) were measured. The serum concentration of nerve growth factor (NGF) was measured.

RESULTSMetoprolol increased DET of ischemic and anoxic myocardium in diabetic rats. The ablation of the left sympathetic nerve increased VFT of diabetic rats. VFT in metoprolo group was significantly increased compared to diabetes group after ischemia. The concentrations of NGF in diabetic group and metoprolol group were higher than those in sham group. There were no difference in NGF levels between ablation of left sympathetic nerve group and sham group.

CONCLUSIONThe remodeling of sympathetic nerve affects the electrophysiology of ischemic myocardium of diabetic rats. Metoprolol can increase the VFT and decrease the excitation threshold of the ischemic myocardium in diabetic rats.

Animals ; Diabetes Mellitus, Experimental ; physiopathology ; Heart ; drug effects ; physiopathology ; Male ; Metoprolol ; pharmacology ; Myocardial Ischemia ; physiopathology ; Nerve Growth Factor ; blood ; Rats ; Rats, Sprague-Dawley ; Sympathectomy

OBJECTIVETo construct the folic acid deficient model in zebrafish and observe the abnormal cardiac phenotypes, to find the optimal period for supplementing folic acid that can most effectively prevent the heart malformation induced by folic acid deficiency, and to investigate the possible mechanisms by which folic acid deficiency induces malformations of heart.

METHODThe folic acid deficient zebrafish model was constructed by using both the folic acid antagonist methotrexate (MTX) and knocking-down dhfr (dihydrofolate reductase gene). Exogenous tetrahydrofolic acid rescue experiment was performed. Folic acid was given to folic acid deficient groups in different periods. The percent of cardiac malformation, the cardiac phenotypes, the heart rate and the ventricular shortening fraction (VSF) were recorded. The out flow tract (OFT) was observed by using fluorescein micro-angiography. Whole-mount in situ hybridization and real-time PCR were performed to detect vmhc, amhc, tbx5 and nppa expressions.

RESULTAbout (78.00 ± 3.74)% embryos in MTX treated group and (68.00 ± 6.32)% embryos in dhfr knocking-down group had heart malformations, including the abnormal cardiac shapes, the hypogenesis of OFT and the reduced heart rate and VSF. Giving exogenous tetrahydrofolic acid rescued the above abnormalities. Given the folic acid on 8 - 12 hours post-fertilization (hpf), both the MTX treated group (20.20% ± 3.77%) and dhfr knocking-down group (43.40% ± 4.51%) showed the most significantly reduced percent of cardiac malformation and the most obviously improved cardiac development. In folic acid deficient group, the expressions of tbx5 and nppa were reduced while the expressions of vmhc and amhc appeared normal. After being given folic acid to MTX treated group and dhfr knocking-down group, the expressions of tbx5 and nppa were increased.

CONCLUSIONSThe synthesis of tetrahydrofolic acid was decreased in our folic acid deficient model. Giving folic acid in the middle period, which is the early developmental stage, can best prevent the abnormal developments of hearts induced by folic acid deficiency. Folic acid deficiency did not disrupt the differentiations of myosins in ventricle and atrium. The cardiac malformations caused by folic acid deficiency were related with the reduced expressions of tbx5 and nppa.

Animals ; Atrial Natriuretic Factor ; metabolism ; Cell Differentiation ; drug effects ; Folic Acid ; metabolism ; Folic Acid Deficiency ; genetics ; metabolism ; Gene Knockdown Techniques ; Heart ; drug effects ; embryology ; growth & development ; T-Box Domain Proteins ; metabolism ; Zebrafish ; embryology ; genetics

OBJECTIVETo evaluate the effect and risk of misoprostol for stimulating cervical maturity in women with post-term pregnancy negative for insulin-like growth factor binding protein-1 (IGFBP-1) in cervical secretion with modified Bishop score less than 3.

METHODSSeventy-one women with post-term pregnancy randomized into misoprostol group (n=37) and control group (n=34) received misoprostol placement at the posterior vaginal fornix and routine intravenous oxytocin infusion, respectively, to stimulate cervical maturity. Failure to respond to the treatment within the initial 24 h necessitated a repeated administration for no more than 3 times in all. Modified Bishop score was recorded and fetal heart monitored once every 24 h, and IGFBP-1 in the cervical secretion was detected at 24 and 48 h after drug administration.

RESULTSThe misoprostol group showed better effect of cervical maturity stimulation than the control group (P<0.001), and the positivity rates of IGFBP-1 24 and 48 h after drug administration were significantly higher than that of the control group (P<0.01 and 0.001). The number of cases with indication for cesarean section was significant higher in the control group (P<0.001). There were no significant differences in postpartum hemorrhage, excessive uterine contraction, incidence of fecal contamination of the amniotic fluid or Apgar score of the newborn between the two groups (P>0.05).

CONCLUSIONSMisoprostol is safe and effective for stimulating cervical maturity in women with post-term pregnancy who have modified Bishop score lower than 3 and are negative for IGPBF-1 in cervical secretion. Oxytocin is not advised for use in such gravida for stimulating cervical maturity. IGFBP-1 in cervical secretion may serve as an important index for evaluating the cervical maturity.

Abortifacient Agents, Nonsteroidal ; administration & dosage ; adverse effects ; therapeutic use ; Administration, Intravaginal ; Adult ; Cervical Ripening ; drug effects ; Cervix Uteri ; drug effects ; metabolism ; Female ; Heart Rate, Fetal ; drug effects ; Humans ; Insulin-Like Growth Factor Binding Protein 1 ; metabolism ; Misoprostol ; administration & dosage ; adverse effects ; therapeutic use ; Pregnancy ; Pregnancy, Prolonged ; drug therapy ; Treatment Outcome

OBJECTIVETo investigate the protective effect and its underlying mechanism of 2,4-diamino-6-hydroxy-pyrimidine (DAHP), an inhibitor of GTP-cyclohydrolase I (GTP-CHI), on postburn Staphylococcus aureus (S. aureus) sepsis in rats.

METHODSFifty-six Wistar rats were randomly divided into four groups, i.e. normal control, scalding control, postburn sepsis group and DAHP treatment group. Tissue samples from liver, kidneys, lungs and heart were aseptically taken, and in which the GTP-CHI and inducible nitric oxide synthase (iNOS) contents and the mRNA expression of tumor necrosis factor-alpha (TNFalpha) were determined. Furthermore, biopterin (BH(4)) and nitric oxide (NO) levels in these tissue were also measured.

RESULTSAfter the scalding injury followed by bacterial challenge, the GTP-CHI gene expression and biopterin levels were significantly increased in all tissue sampled, and so were iNOS mRNA expression and NO (P < 0.01), especially in liver and lungs. The expressions of GTP-CHI mRNA and iNOS mRNA and the production of BH(4) and NO in all tissue were evidently inhibited by the pretreatment with DAHP (P < 0.05 approximately 0.01). At the same time, the TNFalpha expression was also obviously decreased. In addition, The mortality at 6 hr in rats of DAHP treatment group was decreased.

CONCLUSIONThe prognosis of the scalding rats complicated by sepsis caused by G(+) bacteria could be improved by DAHP pretreatment, which might be related to the inhibition of the production of BH(4) and NO by DAHP.

Animals ; Biopterin ; metabolism ; Burns ; complications ; genetics ; metabolism ; Enzyme Inhibitors ; pharmacology ; GTP Cyclohydrolase ; antagonists & inhibitors ; genetics ; Gene Expression Regulation ; drug effects ; Heart ; drug effects ; Hypoxanthines ; pharmacology ; Kidney ; drug effects ; metabolism ; Liver ; drug effects ; metabolism ; Lung ; drug effects ; metabolism ; Male ; Myocardium ; metabolism ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; genetics ; RNA, Messenger ; drug effects ; genetics ; metabolism ; Rats ; Rats, Wistar ; Sepsis ; etiology ; prevention & control ; Staphylococcal Infections ; etiology ; prevention & control ; Staphylococcus aureus ; drug effects ; growth & development ; Sugar Acids ; Time Factors ; Tumor Necrosis Factor-alpha ; genetics

Oxidative stress and apoptosis are the key factors that limit the hypothermic preservation time of donor hearts to within 4-6 h. The aim of this study was to investigate whether the histone deacetylase 3 (HDAC3) inhibitor RGFP966 could protect against cardiac injury induced by prolonged hypothermic preservation. Rat hearts were hypothermically preserved in Celsior solution with or without RGFP966 for 12 h followed by 60 min of reperfusion. Hemodynamic parameters during reperfusion were evaluated. The expression and phosphorylation levels of mammalian STE20-like kinase-1 (Mst1) and Yes-associated protein (YAP) were determined by western blotting. Cell apoptosis was measured by the terminal deoxynucleotidyl-transferase (TdT)-mediated dUTP nick-end labeling (TUNEL) method. Addition of RGFP966 in Celsior solution significantly inhibited cardiac dysfunction induced by hypothermic preservation. RGFP966 inhibited the hypothermic preservation-induced increase of the phosphorylated (p)-Mst1/Mst1 and p-YAP/YAP ratios, prevented a reduction in total YAP protein expression, and increased the nuclear YAP protein level. Verteporfin (VP), a small molecular inhibitor of YAP-transcriptional enhanced associate domain (TEAD) interaction, partially abolished the protective effect of RGFP966 on cardiac function, and reduced lactate dehydrogenase activity and malondialdehyde content. RGFP966 increased superoxide dismutase, catalase, and glutathione peroxidase gene and protein expression, which was abolished by VP. RGFP966 inhibited hypothermic preservation-induced overexpression of B-cell lymphoma protein 2 (Bcl-2)-associated X (Bax) and cleaved caspase-3, increased Bcl-2 mRNA and protein expression, and reduced cardiomyocyte apoptosis. The antioxidant and anti-apoptotic effects of RGFP966 were cancelled by VP. The results suggest that supplementation of Celsior solution with RGFP966 attenuated prolonged hypothermic preservation-induced cardiac dysfunction. The mechanism may involve inhibition of oxidative stress and apoptosis via inactivation of the YAP pathway.
Acrylamides/pharmacology* ; Animals ; Apoptosis/drug effects* ; Cryopreservation ; Disaccharides/pharmacology* ; Electrolytes/pharmacology* ; Glutamates/pharmacology* ; Glutathione/pharmacology* ; Heart/physiology* ; Heart Transplantation/methods* ; Hepatocyte Growth Factor/antagonists & inhibitors* ; Histidine/pharmacology* ; Histone Deacetylase Inhibitors/pharmacology* ; Intracellular Signaling Peptides and Proteins/antagonists & inhibitors* ; Male ; Mannitol/pharmacology* ; Oxidative Stress/drug effects* ; Phenylenediamines/pharmacology* ; Proto-Oncogene Proteins/antagonists & inhibitors* ; Rats ; Rats, Sprague-Dawley ; Signal Transduction/drug effects* ; YAP-Signaling Proteins

OBJECTIVETo evaluate the influence of Chrysanthemum indium on collagen accumulation and signaling transduction pathways in left ventricle tissue of cardiac hypertrophy induced by abdominal aortic banding in rats.

METHODVentricular remodeling was induced by abdominal aortic banding (AAB) in rats. After 35 day treatment, the blood pressure was measured, then the ratios of LVW/BW and HW/BW were calculated. The histological assay was performed by HE staining for determining the myocardium cell cross section and picric acid/sirius red staining for determining collagen content. Immunohistochemistry was used to detect the protein expressions of PKC, bFGF and P38.

RESULTThe experimental data demonstrated that C. indium could decrease blood pressure and the cardiac indexes of LVW/BW and HW/BW, significantly diminish cross sectional area of cardiomyocyte, ameliorate collagen accumulation such as collagen volume fraction, perivascular collagen area and collagen distributions of type I and II and significantly down regulate the protein expressions of PKC, bFGF and P38 (P<0.05).

CONCLUSIONC. indium can significantly attenuate the experimental ventricular remodeling. The mechanism may be related to reducing the blood pressure, decreasing the total collagen content of left ventricle tissue and modulating signaling transduction pathway.

Animals ; Blood Pressure ; drug effects ; Cardiomegaly ; drug therapy ; metabolism ; Chrysanthemum ; Collagen ; metabolism ; Fibroblast Growth Factor 2 ; analysis ; Heart Ventricles ; metabolism ; Male ; Phytotherapy ; Plant Extracts ; pharmacology ; Protein Kinase C ; analysis ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; Ventricular Remodeling ; drug effects ; p38 Mitogen-Activated Protein Kinases ; analysis