title: Brazilein-induced contraction of rat arterial smooth muscle involves activation of Ca2+ entry and ROK, ERK pathways.
abstract: Brazilein (6a,7-dihydro-3,6a,10-trihydroxy-benz[b]indeno[1,2-d]pyran-9(6H)-one) is a compound isolated from Caesalpinia sappan. The vasoactivities of brazilein were evaluated in isolated rat thoracic aorta. The results showed that brazilein can dose-dependently induce contraction of rat thoracic aorta in the resting and phenylephrine pre-evoked state. The average response to 100 microM of brazilein was 30% of the 50 mM KCl contraction, 26% of the 10 muM phenylephrine and 116% of the 20 mM caffeine contraction in comparison. The effects of vasocontraction were proved not to be endothelial dependent and could not be inhibited by alpha-adrenergic receptor blocker phentolamine, beta-adrenergic receptor blocker propranolol, M-adrenaline receptor blocker atropine, angiotensin II receptor blocker losartan or the non-selective nitric oxide synthase (NOS) inhibitor NG-Nitro-L-Arginine Methyl Ester (L-NAME). However the influx of extracellular calcium seemed to be required for this action, because depletion of extracellular calcium and the addition of L-type calcium ion channel antagonist (nimodipine and diltiazem), calcium ion channel activator (BAY-K8644) and potassium ion channel opener (pinacidil) could significantly affect the contraction induced by brazilein. We also investigated the possible signal mechanisms underlying brazilein-induced contraction using selective inhibitors. The inhibitors of myosin light chain kinase (MLCK), Rho-kinase (ROK) and extracellular signal regulated kinase (ERK) can suppress the effect of brazilein respectively, whereas inhibitors of other signaling or receptor molecules such as protein kinase C (PKC) and inositol 1,4,5-triphosphate (IP3) receptor had no effect. All these results demonstrated that brazilein can induce contraction of rat aorta, that the Ca2+ influx, ROK and ERK signal pathways and MLCK activation must be involved in the contractile processes.
authors.affiliation: Protein Science Laboratory of the Ministry of Education, Laboratory of Pharmaceutical Sciences, Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing 100084, China.
authors.forename: Jia
authors.initials: J
authors.lastname: Shen
authors.forename: Siewting
authors.initials: S
authors.lastname: Yip
authors.forename: Zhixing
authors.initials: Z
authors.lastname: Wang
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.forename: Dongming
authors.initials: D
authors.lastname: Xing
authors.forename: Lijun
authors.initials: L
authors.lastname: Du
mesh_terms: D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester / Q000494:pharmacology; D000818:Animals; D001013:Aorta, Thoracic / Q000187:drug effects / Q000502:physiology; D001578:Benzopyrans / Q000737:chemistry / Q000494:pharmacology*; D029911:Caesalpinia / Q000737:chemistry; D002110:Caffeine / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D004110:Diltiazem / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism*; D066298:In Vitro Techniques; D007192:Indenes / Q000737:chemistry / Q000494:pharmacology*; D008297:Male; D015394:Molecular Structure; D009119:Muscle Contraction / Q000187:drug effects*; D009131:Muscle, Smooth, Vascular / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D009553:Nimodipine / Q000494:pharmacology; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors / Q000378:metabolism; D010656:Phenylephrine / Q000494:pharmacology; D020110:Pinacidil / Q000494:pharmacology; D011189:Potassium Chloride / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D015398:Signal Transduction / Q000187:drug effects; D014661:Vasoconstriction / Q000187:drug effects; D054460:rho-Associated Kinases / Q000378:metabolism*
chemical_list: D001578:Benzopyrans; D007192:Indenes; D010656:Phenylephrine; D002110:Caffeine; D009553:Nimodipine; C050295:brazilein; D011189:Potassium Chloride; D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester; D020110:Pinacidil; D019001:Nitric Oxide Synthase; D054460:rho-Associated Kinases; D048049:Extracellular Signal-Regulated MAP Kinases; D004110:Diltiazem; D002118:Calcium; D019331:NG-Nitroarginine Methyl Ester