The cellular mechanisms that contribute to the acceleration of atherosclerosis in diabetes are poorly understood. Therefore, the potential mechanisms involved in the diabetes-dependent increase in vascular smooth muscle cell (VSMC) proliferation was investigated. Using primary culture of VSMC from streptozotocin-induced diabetic rat aorta, cell proliferation assay showed two-fold increase in cell number accompanied with enhanced superoxide generation compared to normal VSMC, 2 days after plating. Both the increased superoxide production and cell proliferation in diabetic VSMC were significantly attenuated by not only tiron (1 mM), a superoxide scavenger, but also by diphenyleneiodonium (DPI; 10micrometer), an NAD (P) H oxidase inhibitor. NAD (P) H oxidase activity in diabetic VSMC was significantly higher than that in control cell, accompanied with increased mRNA expression of p22phox, a membrane subunit of oxidase. Furthermore, inhibition of p22phox expression by transfection of antisense p22phox oligonucleotides into diabetic VSMC resulted in a decrease in superoxide production, which was accompanied by a significant inhibition of cell proliferation. Based on these results, it is suggested that diabetes-associated increase in NAD (P) H oxidase activity via enhanced expression of p22phox contributes to augmented VSMC proliferation in diabetic rats.
1,2-Dihydroxybenzene-3,5-Disulfonic Acid Disodium Salt ; Acceleration ; Animals ; Aorta ; Atherosclerosis ; Cell Count ; Cell Proliferation* ; Membranes ; Muscle, Smooth, Vascular* ; NAD* ; Oligonucleotides ; Oxidoreductases* ; Rats ; RNA, Messenger ; Superoxides ; Transfection

PURPOSE: Many gene polymorphisms are associated with coronary artery abnormalities in Kawasaki disease. Catechol-O-methyltransfe rase (COMT) plays an important role in the metabolism of catecholamines, catechol estrogen, and catechol drugs. Polymorphisms of the COMT gene are reported to be associated with myocardial infarction and coronary artery abnormalities. The aim of this study was to evaluate the relationship between COMT gene polymorphisms and coronary artery abnormalities in Kawasaki disease patients. METHODS: One hundred and one Korean children with Kawasaki disease and 306 healthy Korean control subjects were enrolled in this study. The polymorphisms of the COMT gene were analyzed by direct sequencing. RESULTS: There were no differences in the genotype and allelic frequency of the rs4680 and rs769224 polymorphic sites between Kawasaki disease and control subjects. Further, no significant difference was found in the rs4680 polymorphism between patients with coronary artery abnormalities and patients without coronary artery abnormalities (codominant P=0.32, dominant P=0.74, recessive P=0.13). However, the distribution of the rs769224 polymorphism was significantly different between patie nts with coronary artery abnormalities and patients without coronary artery abnormalities (codominant P=0.0077, dominant P=0.0021, recessive P=0.16). CONCLUSION: Our results indicate that the polymorphisms of the rs769224 gene might be related to the development of coronary artery abnormalities in Kawasaki disease.
Catechol O-Methyltransferase ; Catecholamines ; Catechols ; Child ; Coronary Artery Disease ; Coronary Vessels ; Estrogens ; Genotype ; Humans ; Mucocutaneous Lymph Node Syndrome ; Myocardial Infarction ; Polymorphism, Genetic

BACKGROUND: The intravenous administration of indigo carmine has been reported to produce transiently increased blood pressure in patients. The goal of this in vitro study was to examine the effect of indigo carmine on phenylephrine-induced contractions in an isolated rat aorta and to determine the associated cellular mechanism with particular focus on the endothelium-derived vasodilators. METHODS: The concentration-response curves for phenylephrine were generated in the presence or absence of indigo carmine. Phenylephrine concentration-response curves were generated for the endothelium-intact rings pretreated independently with a nitric oxide synthase inhibitor, Nomega-nitro-L-arginine methyl ester (L-NAME), a cyclooxygenase inhibitor, indomethacin, and a low-molecular-weight superoxide anion scavenger, tiron, in the presence or absence of indigo carmine. The fluorescence of oxidized dichlorofluorescein was measured in rat aortic vascular smooth muscle cells cultured in the control, indigo carmine alone and tiron plus indigo carmine. RESULTS: Indigo carmine (10(-5) M) increased the phenylephrine-induced maximum contraction in the endothelium-intact rings with or without indomethacin, whereas indigo carmine produced a slight leftward shift in the phenylephrine concentration-response curves in the endothelium-denuded rings and L-NAME-pretreated endothelium-intact rings. In the endothelium-intact rings pretreated with tiron (10(-2) M), indigo carmine did not alter phenylephrine concentration-response curves significantly. Indigo carmine (10(-5) M) increased the fluorescence of oxidized dichlorofluorescein in the vascular smooth muscle cells, whereas tiron abolished the indigo carmine-induced increase in oxidized dichlorofluorescein fluorescence. CONCLUSIONS: Indigo carmine increases the phenylephrine-induced contraction mainly through an endothelium-dependent mechanism involving the inactivation of nitric oxide caused by the increased production of reactive oxygen species.
1,2-Dihydroxybenzene-3,5-Disulfonic Acid Disodium Salt ; Administration, Intravenous ; Animals ; Aorta ; Blood Pressure ; Contracts ; Fluorescence ; Humans ; Indigo Carmine ; Indoles ; Indomethacin ; Muscle, Smooth, Vascular ; Nitric Oxide ; Nitric Oxide Synthase ; Phenylephrine ; Prostaglandin-Endoperoxide Synthases ; Rats ; Reactive Oxygen Species ; Superoxides

This study was purpose to investigate the role of reactive oxygen species (ROS) in apoptosis and autophagy induced by FTY720 in multiple myeloma cell line U266. U266 cells were treated by different concentrations of FTY720 for 24 h, the apoptotic rates were detected by flow cytometry, and the expression of LC3B was detected by Western blot. The results indicated that apoptosis and autophagy were induced by FTY720 in U266 cells. Autophagy induced by FTY720 could lead to cell death. Bafilomycin A1, the inhibitor of autophagy, could enhance the cell viability in U266 cells treated with FTY720. NAC or Tiron, ROS scavenger, could decrease the FTY720 induced apoptosis and the expression of LC3B-II was reduced in combination of FTY720 with NAC or Tiron as compared with treatment with FTY720 only. It is concluded that FTY720 can induce U266 cell apoptosis and autophagy. ROS is the mediator that regulates both the apoptosis and autophagy in multiple myeloma cells.
1,2-Dihydroxybenzene-3,5-Disulfonic Acid Disodium Salt ; Apoptosis ; drug effects ; Autophagy ; drug effects ; Cell Line, Tumor ; Fingolimod Hydrochloride ; Humans ; Macrolides ; Microtubule-Associated Proteins ; metabolism ; Multiple Myeloma ; metabolism ; pathology ; Propylene Glycols ; pharmacology ; Reactive Oxygen Species ; metabolism ; Sphingosine ; analogs & derivatives ; pharmacology

Plumbagin, a hydroxy 1,4-naphthoquinone compound from plant metabolites, exhibits anticancer, antibacterial, and antifungal activities via modulating various signaling molecules. However, its effects on vascular functions are rarely studied except in pulmonary and coronary arteries where NADPH oxidase (NOX) inhibition was suggested as a mechanism. Here we investigate the effects of plumbagin on the contractility of skeletal artery (deep femoral artery, DFA), mesenteric artery (MA) and renal artery (RA) in rats. Although plumbagin alone had no effect on the isometric tone of DFA, 1 µM phenylephrine (PhE)-induced partial contraction was largely augmented by plumbagin (ΔT(Plum), 125% of 80 mM KCl-induced contraction at 1 µM). With relatively higher concentrations (>5 µM), plumbagin induced a transient contraction followed by tonic relaxation of DFA. Similar biphasic augmentation of the PhE-induced contraction was observed in MA and RA. VAS2870 and GKT137831, specific NOX4 inhibitors, neither mimicked nor inhibited ΔT(Plum) in DFA. Also, pretreatment with tiron or catalase did not affect ΔT(Plum) of DFA. Under the inhibition of PhE-contraction with L-type Ca²⁺ channel blocker (nifedipine, 1 µM), plumbagin still induced tonic contraction, suggesting Ca²⁺-sensitization mechanism of smooth muscle. Although ΔT(Plum) was consistently observed under pretreatment with Rho A-kinase inhibitor (Y27632, 1 µM), a PKC inhibitor (GF 109203X, 10 µM) largely suppressed ΔT(Plum). Taken together, it is suggested that plumbagin facilitates the PKC activation in the presence of vasoactive agonists in skeletal arteries. The biphasic contractile effects on the systemic arteries should be considered in the pharmacological studies of plumbagin and 1,4-naphthoquinones.
1,2-Dihydroxybenzene-3,5-Disulfonic Acid Disodium Salt ; Animals ; Arteries* ; Catalase ; Coronary Vessels ; Femoral Artery ; Mesenteric Arteries ; Muscle, Smooth ; NADPH Oxidase ; Phenylephrine ; Plants ; Protein Kinase C ; Rats* ; Relaxation ; Renal Artery ; Vasoconstrictor Agents

Tetrahydropapaveroline (THP), a neurotoxic tetrahydroisoquinoline alkaloid formed by condensation between dopamine and dopaldehyde, has been speculated to cause Parkinson's disease and also to contribute to alcohol dependence. Having two catechol moieties, THP may readily undergo oxidation to form an o-quinone intermediate with concomitant production of reactive oxygen species, which can cause neuronal cell death and DNA damage. This review will deal with the current knowledge of neurotoxic effects of this endogenous alkaloid and underlying biochemical mechanisms.
Alcoholism ; Catechols ; Cell Death ; DNA Damage ; Dopamine ; Neurons ; Parkinson Disease ; Reactive Oxygen Species ; Tetrahydroisoquinolines ; Tetrahydropapaveroline

OBJECTIVE: Catechol-O-methyltransferase(COMT) is an important enzyme that inactivates biologically active or toxic catechols. Abnormal catecholamine transmission has been implicated in the pathogenesis of schizophrenia and bipolar disorder. Polymorphism(Val/Met) of the COMT gene was shown to determine high-and low-activity alleles of the enzyme. This study was designed to investigate the association between COMT gene polymorphism and schizophrenia and bipolar disorder in a Korean population. METHOD: COMT gene were genotyped with polymerase chain reaction and restriction enzyme NlaIII in 128 patients with schizophrenia, 110 with bipolar disorder, and 176 controls. RESULTS: 1) The distribution of the COMT genotype in schizophrenic patients with Val/Val, Val/Met, Met/Met were 76(59.4%), 43(33.6%), 9(7.0%), in bipolar disorder patients were 63(57.3%), 35(31.8%), 12(10.9%), and in the controls were 83(47.2%), 79(44.9%), 14(8.0%). The allele frequencies of the COMT gene in schizophrenic patients with Val and Met were 195(76.2%), 61(23.8%), in bipolar disoreder patients were 161(73.2%), 59(26.8%), and in the controls were 245(69.6%), 107(30.4%). 2) There were no differences in genotype distribution and allele frequencies of COMT gene polymorphism among the 3 groups. Neither patients with schizophrenia nor bipolar disorder differed in the genotype and allelic frequencies from the controls. CONCLUSION: These results suggest COMT gene polymorphism is not causally related to the development of schizophrenia and bipolar disorder in a Korean Population.
Alleles ; Bipolar Disorder* ; Catechol O-Methyltransferase* ; Catechols ; Gene Frequency ; Genotype ; Humans ; Polymerase Chain Reaction ; Schizophrenia*

Catechols ; urine ; Chromatography, High Pressure Liquid ; methods ; Electrochemistry ; methods ; Humans ; Hydroquinones ; urine

OBJECTIVETo discuss the synergistic mechanism of compatible use of two medicinal herbs, Zingiber offiicinale and Aconitum cainichaeli, by determining single decoction of Z. offiicinale and four gingerols (6-gingerol, 8-gingerol, 6-shogaol, 10-gingerol) contained in compound decoction of Z. offiicinale and A. cainichaeli of different compatibility ratio using HPLC.

METHODKromasil-C18 column (4.6 mm x 250 mm, 5 microm) was adopted. The mobile phase was acetonitrile (B) and 0.1% aqueous acetic acid (A) for gradient elution (0-30 min, 40%-90% B; 30-35 min, 90%-40% B). The flow rate was 1.0 mL x min(-1). The detection wavelength was set at 275 nm. The column temperature was 30 degrees C.

RESULTThe four gingerols were in baseline separation, with a good linearity (r > 0.999), an average recovery of 100.9% -103.5% and RSD < 3.0%. Compared with the single decoction of Z. offiicinale, the content of gingerols in the compound decoction of Z. offiicinale and A. cainichaeli was on the rise and in direct proportion with the increase in the volume of A. cainichaeli.

CONCLUSIONThe synergistic mechanism of the compatibility of Z. offiicinale and A. cainichaeli can be proved with the increased release of gingerols from Z. offiicinale.

Plantamajoside is one of the main bioactive compounds in Plantaginis Herba A TLC method was developed identification of plantamajoside in 11 Plantaginis Herba samples using silica gel G as coating substance and a mixture of ethyl acetiate methanol-formic acid-water (18: 3 : 1.5 : 1) as a developing solvent, the established TLC condition displayed a very well separation on the chromatogram of tested Plantaginis Herba samples and the marker compound plantamajoside showed as a distinct light-blue fluorescence spot observed under UV 365 nm. Using the HPLC method, plantamajoside was separated at 30 degrees C on a Promocil C18, (4.6 mm x 250 mm, 5 microm) column with acetonitrile-0.1% formic acid (17:83) as the mobile phase. The detection wavelength was set at 330 nm and the flow rate was 1 mL x min(-1). The calibration curve of plantamajoside displayed ideal linearity over the range of 0.0499-11.9664 microg (r = 0.9999), and the average recovery of plantamajoside was 100.6% with a RSD of 2.7%. The contents of plantamajoside were in the range of 0.067%-1.80% in Plantaginis Herba The established TLC identification and HPLC were sensitive, reliable and repeatable, which can be applied for the quality evaluation and standard criteria of Plantaginis Herba.
Asteraceae ; chemistry ; Catechols ; analysis ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Glucosides ; analysis ; Reproducibility of Results