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20-HETE-Induced Changes in Inotropic Effects Depend on the Activation or Inactivation of L-type Calcium Channels by PKA and\or PKC in Rat Ventricular Myocytes
Author: LiXingZuo
Tutor: ZengQingHua
School: Northeast Normal University
Course: Physiology
Keywords: 20-HETE Ventricular myocytes L- type calcium channel currents Intracellular calcium ion concentration Calcium transient PKA PKC
CLC: Q26
Type: Master's thesis
Year: 2009
Downloads: 55
Quote: 0
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Abstract
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Cytochrome P450s (CYP) ω- hydroxylase arachidonic acid metabolites 20 - arachidonic acid hydroxyalkyl eicosane (20-HETE), has an important role in regulation of vascular tone , but the 20-HETE on the heart the role has rarely been reported . We studied the effects of exogenous 20-HETE on myocardial contractility regulation and its mechanism. Using the isolated perfused Langendorff systematically studied 20-HETE on isolated cardiac effects of myocardial mechanics , found that low doses of 20-HETE (0.1-3 nmol / L) on the perfused heart has a positive inotropic effect , whereas higher doses ( 10 nmol / L) is a clear negative effect. Then we use Fura-2/AM fluorescence measurement and patch clamp techniques to study the 20-HETE in rat ventricular myocytes isolated intracellular calcium concentration ([Ca2] i) and L- type calcium channel effect. We found that low doses of 20-HETE (0.01, 0.03, 0.1μmol / L) increased in a dose -dependent manner [Ca2] i, and high doses of 20-HETE (0.3, 1μmol / L) induced [Ca2] i in reduced. And that low doses of 20-HETE (0.1μmol / L) induced by whole cell L- type calcium channel current amplitude increased, while the high dose of 20-HETE (1μmol / L) has the opposite effect . With a laser scanning confocal microscope line function testing 20-HETE on intracellular calcium transients , found that low concentrations of 20-HETE (0.01,0.1 μmol / L) can increase the amplitude of calcium transient and prolonged calcium transient decline phase schedule ; high concentrations of 20-HETE (1.0μmol / L) can reduce the calcium transient amplitude and prolong the time course of calcium transients . The 20-HETE -induced [Ca2] i, whole-cell L- type calcium channel currents , calcium transient increase in the amplitude of such effects may be protein kinase C (PKC) inhibitor chelerythrine base and cAMP-dependent protein kinase (PKA) inhibitor H-89 block. PKC and PKA activity measurement results show that 20-HETE (0.03μmol / L) increased cardiac activity of PKC and PKA , and the red base celandine (5μmol / L) , and H-89 (5μmol / L) inhibited this effect . These data suggest that 20-HETE may activate or inhibit L-type calcium channels to regulate [Ca2] i, thereby adjusting the force of myocardial contraction , and these effects mediated by the PKC and PKA .
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CLC: > Biological Sciences > Cell Biology > Cellular Biochemistry
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