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The modulatory effect of the protein kinase C activator was examined on contraction of rat isolated vas deferens induced by constrictive agonists, noradrenaline (NA), ATP, BaCl2 and high K+. Phorbol 12,13-diacetate (PDA, 1 micromol/l) induced a transient extracellular Ca(2+)-dependent contraction while the inactive analogue, 4alpha-phorbol (1 micromol/l) had no effect. PDA significantly enhanced the peak amplitude of the contractile response to NA (0.1-10 micromol/l), ATP (100 micromol/l), Ba2+ (3 mmol/l) or high K+ (30 mmol/l). Staurosporine at 30 nmol/l reduced the enhancing effect of PDA on the agonist-induced contraction. NA (10 micromol/l) produced a phasic contraction followed by a sustained contraction, while ATP induced monophasic contraction. Pretreatment with nifedipine (10 nmol/l) had no effect on the phasic contraction induced by NA, but it significantly reduced ATP- or high K(+)-induced contraction. Staurosporine (30 nmol/l) alone attenuated the peak contractile response induced by NA or ATP but not by Ba2+. NA produced a transient contraction in Ca(2+)-free Krebs solution, and PDA (1 micromol/l) markedly enhanced this effect. These novel data indicate that activation of a protein kinase C-dependent mechanism not only affects contraction mediated by Ca2+ influx through voltage-sensitive Ca2+ channels, but also promotes intracellular Ca2+ release or intracellular Ca(2+)-mediated contractile mechanism in rat vas deferens., (Copyright 2001 S. Karger AG, Basel)

Aim: To examine the role of endothelium in the vascular responses to flavonoids, baicalein, baicalin, cardamonin, alpinetin, and to purified jasmine green tea (-)epicatechin in the isolated rate mesenteric artery rings., Methods: The isometric contraction was measured by Grass force-displacement transducers., Results: Both baicalein and baicalin enhanced the phenylephrine-induced contractile response in the endothelium-intact rings. This enhancement was abolished by pretreatment with the nitric oxide inhibitor NG-nitro-L-arginine or in the absence of the endothelium. Both flavonoids also inhibited the acetylcholine-induced endothelial nitric oxide-dependent relaxation. In contrast, cardamonin, alpinetin or (-)epicatechin induced both endothelium-dependent and -independent relaxation. NG-nitro-L-arginine meyhyl ester or endothelium denudation attenuated the endothelium-dependent relaxation to the same extent., Conclusion: Baicalein and baicalin enhanced the phenylephrine-induced contraction most likely through inhibiting production or/and release of endothelial nitric oxide. Whilst, cardamonin-, alpinetin- or (-)epicatechin-induced endothelium-dependent relaxation is primarily mediated through endothelial nitric oxide.

Aim: The present study was aimed to examine the vasorelaxant and antiproliferative responses to purified green tea epicatechin mixture, (-)epicatechin and (-)epigallocatechin gallate on rat arterial smooth muscle cells., Methods: Changes in isometric force were measured by Grass force transducer and effects on cell proliferation were evaluated using [3H]thymidine incorporation assay., Results: Epicatechin mixture, (-)epicatechin and (-)epigallocatechin gallate, which we isolated and purified from jasmine green tea, concentration-dependently, reduced the contractile response to phenylephrine in rat isolated aortic rings with (-)epigallocatechin gallate being more effective. These three agents also inhibited [3H]thymidine incorporation into DNA in cultured rat aortic smooth muscle cells in a concentration-dependent manner., Conclusion: The purified epicatechin derivatives from jasmine green tea relaxed the isolated rat arteries preconstricted by phenylephrine and inhibited aortic smooth muscle cell proliferation.

The endothelium-dependent and -independent relaxant effect of procaine was examined in isolated rat aortic rings. Procaine induced relaxation of arteries precontracted with phenylephrine or with 60 mM K+ in a concentration-dependent manner (0.01-3 mM). Procaine (1 mM) inhibited the transient contraction induced by caffeine (10 mM) in Ca2+-free Krebs solution. Removal of the endothelium caused a rightward shift of the concentration-response curve for procaine. N(G)-Nitro-L-arginine (L-NNA, 10-100 microM), N(G)-nitro-L-arginine methyl ester (L-NAME, 100 microM) and methylene blue (1-10 microM) significantly attenuated the procaine-induced relaxation without affecting the maximal response. L-Arginine (1 mM) partially but significantly antagonized the effect of L-NAME (100 microM). Pretreatment of endothelium-intact aortic rings with procaine (1 mM) or with acetylcholine (10 microM) significantly elevated the tissue contents of cyclic GMP and this increase was inhibited in the presence of 100 microM L-NNA. Tetrapentylammonium ions (1-3 microM) reduced the procaine-induced relaxation in both endothelium-intact and -denuded arteries. Tetrapentylammonium ions (3 microM) did not affect the procaine-induced relaxation of 60 mM K+-contracted arteries. Tetraethylammonium ions (3 mM) inhibited the procaine-induced relaxation. In contrast, iberiotoxin (100 nM), glibenclamide (3 microM), 4-aminopyridine (3 mM) and indomethacin (10 microM) had no effect. These results indicate that the procaine-induced relaxation may be mediated through multiple mechanisms. A substantial portion of the procaine-induced relaxation in rat aorta was caused by nitric oxide but not by other endothelium-derived factors. The activation of tetrapentylammonium- and tetraethylammonium-sensitive K+ channels contributes in part to the procaine-induced vasorelaxation. Besides, procaine may directly inhibit both external Ca2+ entry and internal Ca2+ release in aortic smooth muscle cells.

The modulatory effect of the protein kinase C activator was examined on contraction of rat isolated vas deferens induced by constrictive agonists, noradrenaline (NA), ATP, BaCl[sub 2] and high K[sup +] . Phorbol 12,13-diacetate (PDA, 1 μmol/l) induced a transient extracellular Ca[sup 2+] -dependent contraction while the inactive analogue, 4α-phorbol (1 μmol/l) had no effect. PDA significantly enhanced the peak amplitude of the contractile response to NA (0.1–10 μmol/l), ATP (100 μmol/l), Ba[sup 2+] (3 mmol/l) or high K[sup +] (30 mmol/l). Staurosporine at 30 nmol/l reduced the enhancing effect of PDA on the agonist-induced contraction. NA (10 μmol/l) produced a phasic contraction followed by a sustained contraction, while ATP induced monophasic contraction. Pretreatment with nifedipine (10 nmol/l) had no effect on the phasic contraction induced by NA, but it significantly reduced ATP- or high K[sup +] -induced contraction. Staurosporine (30 nmol/l) alone attenuated the peak contractile response induced by NA or ATP but not by Ba[sup 2+] . NA produced a transient contraction in Ca[sup 2+] -free Krebs solution, and PDA (1 μmol/l) markedly enhanced this effect. These novel data indicate that activation of a protein kinase C-dependent mechanism not only affects contraction mediated by Ca[sup 2+] influx through voltage-sensitive Ca[sup 2+] channels, but also promotes intracellular Ca[sup 2+] release or intracellular Ca[sup 2+] -mediated contractile mechanism in rat vas deferens.Copyright © 2001 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]