Scutellarin (SCU), a flavonoid from a traditional Chinese medicinal plant. Our previous study has demonstrated that SCU relaxes mouse aortic arteries mainly in an endothelium-depend-ent manner. In the present study, we investigated the vasoprotective effects of SCU against HR-induced endothelial dysfunction (ED) in isolated rat CA and the possible mechanisms involving cyclic guanosine monophosphate (cGMP) dependent protein kinase (PKG). The isolated endothelium-intact and endothelium-denuded rat CA rings were treated with HR injury. Evaluation of endothelium-dependent and -independent vasodilation relaxation of the CA rings were performed using wire myography and the protein expressions were assayed by Western blotting. SCU (10-1 000 μmol·L(-1)) could relax the endothelium-intact CA rings but not endothelium-denuded ones. In the intact CA rings, the PKG inhibitor, Rp-8-Br-cGMPS (PKGI-rp, 4 μmol·L(-1)), significantly blocked SCU (10-1 000 μmol·L(-1))-induced relaxation. The NO synthase (NOS) inhibitor, NO-nitro-L-arginine methylester (L-NAME, 100 μmol·L(-1)), did not significantly change the effects of SCU (10-1 000 μmol·L(-1)). HR treatment significantly impaired ACh-induced relaxation, which was reversed by pre-incubation with SCU (500 μmol·L(-1)), while HR treatment did not altered NTG-induced vasodilation. PKGI-rp (4 μmol·L(-1)) blocked the protective effects of SCU in HR-treated CA rings. Additionally, HR treatment reduced phosphorylated vasodilator-stimulated phosphoprotein (p-VASP, phosphorylated product of PKG), which was reversed by SCU pre-incubation, suggesting that SCU activated PKG phosphorylation against HR injury. SCU induces CA vasodilation in an endothelium-dependent manner to and repairs HR-induced impairment via activation of PKG signaling pathway.
Animals ; Apigenin ; pharmacology ; Cell Adhesion Molecules ; drug effects ; Cell Hypoxia ; Coronary Vessels ; drug effects ; Cyclic GMP ; analogs & derivatives ; metabolism ; pharmacology ; Cyclic GMP-Dependent Protein Kinases ; Glucuronates ; pharmacology ; Microfilament Proteins ; drug effects ; NG-Nitroarginine Methyl Ester ; metabolism ; pharmacology ; Phosphoproteins ; drug effects ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; complications ; physiopathology ; Signal Transduction ; drug effects ; Thionucleotides ; metabolism ; pharmacology ; Vasodilation ; drug effects ; physiology

OBJECTIVETo investigate the vasodilative action and the possible mechanisms of Shenmai injection on porcine coronary artery.

METHODIsometric tensions of the porcine coronary artery ring precontracted with potassium chloride (KCl) were recorded in vitro when the doses of 0.1, 0.5, 1, 5, 10, 50 mL x L(-1) of Shenmai injective were cumulatively added into the organ bath of porcine coronary artery ring.

RESULTShenmai injection caused same vasorelaxation of porcine coronary artery rings with endothelium intact and denuded precontracted with KCl in a concentration-dependent manner. Pretreatment with TEA and 4-AP prohibited the vasorelaxation by Shenmai injection, but pretreatment with KB-R7943, BaCl2, Gli did not affect the vascular effect of Shenmai injection.

CONCLUSIONShenmai injection could produce vasodilatation on KCl pre-contracted porcine coronary artery rings. It seems that K(Ca) and K(V) channel play an important role in the relaxation of the porcine coronary artery rings pre-contracted with KCl.

Animals ; Aorta, Thoracic ; drug effects ; physiology ; Coronary Vessels ; drug effects ; physiology ; physiopathology ; Drugs, Chinese Herbal ; pharmacology ; Female ; In Vitro Techniques ; Male ; Nitric Oxide ; metabolism ; Swine ; Vasodilation ; drug effects ; Vasodilator Agents ; pharmacology

OBJECTIVEPeriplocin is an active digitalis-like component from Cortex Periplocae, which has been widely used in the treatment of heart diseases in China for many years. According to the recommendations on the cardiovascular effect of periplocin from in vivo experiments, subsequent in vitro experiments are greatly needed for the global assessment of periplocin. The objective of this study is to investigate the cell proliferation effect and the mechanism of periplocin on endothelial cells.

METHODSThe proliferative activity of periplocin (0.4, 2, 10, 50, 250 micromol/L; 6, 12, 24, 48, 72 h) was investigated by a comparison with the well-reported cardiac glycoside, ouabain, on mouse cardiac microvascular endothelial cells (CMEC). 3-(4,5-dimethylthiazolyl)-2,5-diphenyltetrazolium bromide (MTT), lactate dehydrogenase (LDH) and 5-bromo-2-deoxyuridine (BrdU) assays were used to evaluate cell proliferation and viability. Subsequently, cDNA microarray experiments were performed on periplocin- (50 micromol/L) and ouabain- (50 micromol/L) treated cells, and data was analyzed by ArrayTrack software.

RESULTSPeriplocin could increase cell viability to a level lower than ouabain in the MTT analysis, but decrease LDH release simultaneously. The BrdU incorporation assay showed an increase in cell proliferation with 2-50 micromol/L periplocin. Genes related to protein serine/threonine kinase were the most significantly enriched in the 160 genes identified in periplocin versus the control. In the 165 genes regulated by periplocin versus ouabain, GTP-binding was the most altered term.

CONCLUSIONSThe results demonstrated the proliferation action of periplocin on CMEC. Meanwhile, its lower cytotoxicity compared to ouabain provides a new insight into the treatment of heart failure.

Animals ; Animals, Newborn ; Cardiac Glycosides ; pharmacology ; Cardiotonic Agents ; pharmacology ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; genetics ; Cells, Cultured ; Coronary Vessels ; drug effects ; metabolism ; physiology ; Drug Evaluation, Preclinical ; Endothelial Cells ; drug effects ; metabolism ; physiology ; Gene Expression Profiling ; Gene Expression Regulation ; drug effects ; Mice ; Microvessels ; drug effects ; metabolism ; physiology ; Models, Biological ; Myocardium ; metabolism ; Oligonucleotide Array Sequence Analysis ; Ouabain ; pharmacology ; Saponins ; pharmacology

OBJECTIVEMany clinical evidences and epidemiologic data in the past suggested that Kawasaki disease (KD) is correlated with an acute immune dysfunction caused by infection. In our preliminary study, Toll-like receptor 4 signal pathway, which could activate nuclear transcription factor-kappaB and induce excessive product of proinflammatory cytokines, chemokines and co-stimulatory molecules, was observed to be significantly activated during acute phase of Kawasaki disease. But the causative factors and regulatory mechanism are still unknown. In this study, the authors further investigated the changes and significances of regulatory factors for signal pathway of Toll-like receptors (TLRs) in immunological pathogenesis of Kawasaki disease.

METHODSForty-eight children with KD, sixteen children with infectious disease (ID) and sixteen age-matched healthy children were studied. Reverse-transcription PCR (RT-PCR) and real-time PCR were used to evaluate the expression levels of regulatory and effective factors in toll-like receptor 4 (TLR4) signal pathways and proinflammatory factors in peripheral blood monocyte/macrophage (MC). The expression of TLR4 protein in MC was analyzed by flow cytometry.

RESULTS(1) Expression levels of TLR4, MD-2, MyD88, IRAK-4, TRAF6, TAK1, TAB1 and TAB2 mRNA in KD group were elevated significantly during acute phase (P < 0.05). (2) Transcription levels of regulatory factors PRAT4B and STAP2 in patients with KD or ID were found to be higher than those in the healthy volunteers (P < 0.05), but no significant differences in these parameters were detected between KD patients and ID patients (P > 0.05). Transcription levels of regulatory factors such as FLN29, RP105 and MD-1 were up-regulated to some extents and expression level of DAP12 mRNA in KD patients were found to be lower than that in normal controls (P < 0.05), while all of the four regulatory factors were found to be lower than those in ID patients (P < 0.05). Expressions of proinflammatory cytokines such as L-1beta, IL-6 and TNF-alpha in KD patients were significantly higher than those in ID patients (P < 0.05). (3) Stimulation with lipopolysaccharide (LPS) elevated remarkably the expressions of regulatory factors PRAT4B and STAP2 in KD patients or healthy volunteers (P < 0.05). All of the four negative-regulatory factors were found to be significantly up-regulated after stimulation with LPS in controls (P < 0.05). No responses to LPS were observed in expression of FLN29, RP105 and MD-1 mRNA in KD patients (P > 0.05), except for increased transcription of DAP12. (4) The levels of PRAT4B and STAP2 mRNA in KD patients with coronary artery lesion (KD-CAL(+)) were detected to be higher than those in KD patients without coronary artery lesion (KD-CAL(-)) during acute phase (P < 0.05), while those of FLN29, RP105 and MD-1 in KD-CAL(+) group were lower than that in the latter (P < 0.05). No significant difference in DAP12 mRNA expression level was detected between the two groups (P > 0.05). Expressions of proinflammatory cytokines and TLR4 protein on surface of CD14-positive cells in KD-CAL(+) group were found to be higher than those in KD-CAL(-) group [(11.9 +/- 2.4)% vs. (6.5 +/- 1.7)%, P < 0.05].

CONCLUSIONDisturbance of negative-regulatory factors may be one of the factors causing aberrant immunological function in KD.

Child ; Coronary Vessels ; drug effects ; physiology ; Cytokines ; metabolism ; Flow Cytometry ; Humans ; Leukocytes, Mononuclear ; drug effects ; metabolism ; Lipopolysaccharides ; toxicity ; Macrophages ; drug effects ; pathology ; Mucocutaneous Lymph Node Syndrome ; immunology ; metabolism ; physiopathology ; RNA, Messenger ; blood ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Signal Transduction ; drug effects ; Toll-Like Receptor 4 ; physiology ; Toll-Like Receptors ; immunology ; metabolism ; Tumor Necrosis Factor-alpha ; pharmacology ; Up-Regulation

OBJECTIVETo study the effect of Shumai Capsule (SMC) on angiogenesis and expression of relevant growth factor in rats with myocardial ischemia (MI).

METHODSModel rats of MI were duplicated and treated with SMC (SMC group), bFGF + calparine (positive control group) and normal saline (model group) respectively. Besides, a sham-operative group was set up and treated with normal saline. The rats were sacrificed in batches at the time after being medicated for 1, 2 and 4 weeks, for determining von Willebrand factor (vWF) and vascular endothelial growth factor (VEGF) protein expression in ischemic myocardium by immuno-histochemical staining, myocardial micro-vessel density (MVD) using digital analysis system, and the gene expression of VEGF by quantitative real-time PCR.

RESULTSCompared with the sham-operative group and the model group, levels of MVD, protein and gene expression of VEGF in the SMC group were higher respectively at three time segments (all P <0.01), but showed insignificant difference to those in the positive control group.

CONCLUSIONSMC could promote angiogenesis in ischemic myocardium of rats, the up-regulation on VEGF mRNA and protein expression might be one of the potential mechanisms of SMC in promoting angiogenesis.

Animals ; Capsules ; Coronary Vessels ; drug effects ; physiology ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Gene Expression ; drug effects ; Male ; Myocardial Ischemia ; physiopathology ; Myocardium ; metabolism ; pathology ; Neovascularization, Physiologic ; drug effects ; Phytotherapy ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction ; Vascular Endothelial Growth Factor A ; biosynthesis ; genetics

The aim of the present study was to examine the effects of tetramethylpyrazine (TMP) on large-conductance Ca(2+)-activated potassium channels (BK(Ca) channels) in porcine coronary artery smooth muscle cells, in order to provide the experimental evidence for expounding the mechanism of TMP in dilating coronary artery. Cell-attached and inside-out single channel recording techniques were used to observe the effects of TMP on BK(Ca) channels as well as the effects after the cells were treated by protein kinase A (PKA) inhibitor or protein kinase G (PKG) inhibitor. In inside-out patch, TMP activated BK(Ca) channels by increasing open-state probability (N(Po)) and decreasing close time (Tc) in a concentration-dependent manner. TMP (0.73~8.07 mmol/L) in the bath solution increased N(Po) from (0.01+/-0.003) to (0.03+/-0.01)~(1.21+/-0.18) (P<0.01, n=10), and decreased Tc from (732.33+/-90.67) ms to (359.67+/-41.30) ~ (2.96+/-0.52) ms (P<0.01, n=10). These actions of TMP occurred even when the free Ca(2+) concentration in the bath was reduced to ~ 0 mmol/L. The specific inhibitors of PKA (H-89, 3 mumol/L) and PKG (KT-5823, 1 mumol/L) had no influence on the activation of TMP on BK(Ca) channels. These findings suggest that TMP can directly activate BK(Ca) channels in coronary artery smooth muscle, which probably is an important mechanism in dilating coronary artery.
Animals ; Coronary Vessels ; cytology ; Muscle, Smooth, Vascular ; cytology ; metabolism ; Patch-Clamp Techniques ; Potassium Channels, Calcium-Activated ; drug effects ; physiology ; Pyrazines ; pharmacology ; Swine ; Vasodilator Agents ; pharmacology

OBJECTIVETo assess the effect of beta blocker on blood flow velocity and reserve on the intramural coronary artery of patients with myocardial bridging.

METHODSIn 8 patients with myocardial bridge, intracoronary Doppler was performed before and after esmolol was given intravenously. The basic average peak velocity (bAPV), hyperaemic average peak velocity (hAPV) of blood flow, and coronary flow reserve (CFR) proximal and distal to the mural myocardial bridging was measured and compared.

RESULTSAfter esmolol injection, the mural coronary diameter systolic reduction decreased from (58.0 +/- 14.7)% to (26.0 +/- 9.8)% (P < 0.01); the bAPV proximal and distal to myocardial bridging separately decreased from (19.4 +/- 4.9) cm/s and (18.4 +/- 3.6) cm/s to (4.7 +/- 3.9) cm/s (P < 0.01) and (15.1 +/- 1.5) cm/s (P < 0.05). Under hyperemization, esmolol changed the hAPV of proximal and distal to myocardial bridging separately from (54.1 +/- 14.9) cm/s and (44.7 +/- 9.4) cm/s to (49.7 +/- 16.4) cm/s and (48.9 +/- 10.1) cm/s (all P > 0.05); thus, the value of CFR both proximal and distal to myocardial bridge increased separately from 2.8 +/- 0.3 and 2.5 +/- 0.5 to 3.4 +/- 0.5 and 3.2 +/- 0.6 (all P < 0.01).

CONCLUSIONEsmolol can decreased the compression of the intramural coronary artery and increased the CFR to normal level of it.

Adrenergic beta-Antagonists ; pharmacology ; therapeutic use ; Coronary Circulation ; drug effects ; physiology ; Coronary Vessels ; diagnostic imaging ; drug effects ; physiopathology ; Female ; Humans ; Male ; Middle Aged ; Myocardial Bridging ; drug therapy ; physiopathology ; ultrastructure ; Propanolamines ; pharmacology ; therapeutic use ; Ultrasonography, Interventional

Transthoracic Doppler echocardiography (TTDE) allows noninvasive flow measurement in the distal left anterior descending artery (LAD). The feasibility of detecting coronary flow by contrast-enhanced TTDE with second harmonic technique was assessed, the coronary flow velocity reserve (CFVR) was evaluated in comparison to intracoronary Doppler flow (ICD) analysis and the CFVR after PTCA in LAD was investigated. In 77 (96%) of 80 patients, CFVR was successfully determined with intravenous adenosine infusion. Doppler signal quality was evaluated in the first 46 patients by use of intravenous Levovist infusion and second harmonic technique. The Doppler flow was not visible in 1. patient only. CFVR determined from TTDE (2.77 +/- 0.65) was correlated closely with those from ICD (2.88 +/- 0.78) measurements (y = 0.73x + 0.67, r = 0.87, P < 0.001). In conclusion, TTDE is a feasible method and provides reliable data on CFVR which can be used for follow-up after PTCA.
Adenosine ; administration & dosage ; Adult ; Aged ; Blood Flow Velocity ; Contrast Media ; Coronary Artery Disease ; diagnostic imaging ; physiopathology ; Coronary Circulation ; physiology ; Coronary Vessels ; diagnostic imaging ; drug effects ; physiopathology ; Echocardiography, Doppler ; methods ; Female ; Humans ; Male ; Middle Aged ; Polysaccharides ; Predictive Value of Tests ; Vasodilator Agents ; administration & dosage

The reliability and reliable indexes of quantitative assessment of coronary flow reserve (CFR) by using time-intensity curve (TIC) via myocardial contrast echocardiography were investigated. The TIC variables were obtained by employing acoustic densitometry (AD) technique before and after acetylcholine (Ach) injection in 12 dogs. Meanwhile, the correlation between these variables and CFR was analyzed. Among the variables derived from TIC, peak intensity (PI), area under the curve (AUC) and descending slope (DS) were increased significantly (P < 0.05) with the increase of coronary blood flow after Ach injection. Conversely, time-to-peak (TP), half-time of descent (HT), and mean-transit-time (MTT) were decreased remarkably (P < 0.0001). The PI and AUC ratios from post- to pre-Ach injection were strongly associated with CFR with the correlation coefficient (r) being 0.8366 and 0.8824, respectively. It is reliable by using the variables derived from TIC with myocardial contrast echocardiography to quantitatively evaluate regional myocardial CFR. The PI and AUC ratios from post- to pre-Ach injection are the reliable indexes for quantitative assessment of CFR.
Animals ; Blood Flow Velocity ; physiology ; Contrast Media ; Coronary Circulation ; physiology ; Coronary Vessels ; diagnostic imaging ; physiology ; Dogs ; Echocardiography ; methods ; Image Processing, Computer-Assisted ; Observer Variation ; Regional Blood Flow ; drug effects ; physiology ; Reproducibility of Results ; Ultrasonography, Interventional

The reliability and reliable indexes of quantitative assessment of coronary flow reserve (CFR) by using time-intensity curve (TIC) via myocardial contrast echocardiography were investigated. The TIC variables were obtained by employing acoustic densitometry (AD) technique before and after acetylcholine (Ach) injection in 12 dogs. Meanwhile, the correlation between these variables and CFR was analyzed. Among the variables derived from TIC, peak intensity (PI), area under the curve (AUC) and descending slope (DS) were increased significantly (P < 0.05) with the increase of coronary blood flow after Ach injection. Conversely, time-to-peak (TP), half-time of descent (HT), and mean-transit-time (MTT) were decreased remarkably (P < 0.0001). The PI and AUC ratios from post- to pre-Ach injection were strongly associated with CFR with the correlation coefficient (r) being 0.8366 and 0.8824, respectively. It is reliable by using the variables derived from TIC with myocardial contrast echocardiography to quantitatively evaluate regional myocardial CFR. The PI and AUC ratios from post- to pre-Ach injection are the reliable indexes for quantitative assessment of CFR.
Blood Flow Velocity/physiology ; Contrast Media ; Coronary Circulation/*physiology ; Coronary Vessels/physiology ; Coronary Vessels/ultrasonography ; *Echocardiography/methods ; *Image Processing, Computer-Assisted ; Observer Variation ; Regional Blood Flow/drug effects ; Regional Blood Flow/physiology ; Reproducibility of Results ; Ultrasonography, Interventional