Role of Protein Kinase C in Relationship between Ca2+ and Contractile Elements in Rat .ALPHA.-Toxin-permeabilized Mesenteric Artery.
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Phorbol ester, which activates protein kinase C(PKC), modulates vasoconstrictor-induced tension in vascular smooth muscle. Recently, Staphylococcal aureus α-toxin, which produces too small pores in the plasma membrane to allow passage of proteins, such as PKC, is used to investigate the signal transduction system in vascular smooth muscle cells. In order to elucidate the role of PKC on vascular smooth muscle contraction, we examined whether PKC activation influences the relationship between intracellular Ca<SUP>2+</SUP> ([Ca<SUP>2+</SUP>]<SUB>i</SUB>) and tension in Wistar rat superior mesenteric artery(SMA) using vascular smooth muscle permeabilized with Staphylococcal α-toxin. [Ca<SUP>2+</SUP>]<SUB>i</SUB> was clamped at specified values (10<SUP>-8.5</SUP>-10<SUP>-4</SUP>mol/L) using EGTA-Ca<SUP>2+</SUP> buffer. In α-toxin non-treated rings of SMA, isometric tension was evoked by 10 mmol/L caffeine and 10-30 mmol/L external potassium (high K<SUP>+</SUP>) in the absence or presence of phorbol 12, 13-dibutyrate (PDBu), a PKC activator, 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H-7), and staurosporine (PKC inhibitors). PDBu significantly augmented caffeine- and high K<SUP>+</SUP>-evoked contractions. H-7 and staurosporine significantly attenuated caffeine- and high K<SUP>+</SUP>-evoked contractions augmented by PDBu. Moreover, H-7 significantly suppressed high K<SUP>+</SUP>-induced contraction in the absence of PDBu. In α-toxin permeabilized artery, PDBu shifted the [Ca<SUP>2+</SUP>]<SUB>i</SUB>-force relationship curve to the left. These results suggest that PKC activates vascular smooth muscle contraction by increasing the sensitivity of the contractile apparatus to Ca<SUP>2+</SUP> .
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社団法人 日本循環器学会 | 論文
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