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1 The Second Department of Internal Medicine, School of Medicine, Nagoya University, Nagoya 466-8560; and 2 Department of Humoral Regulation and 3 Department of Circulation, Research Institute of Environmental Medicine, Nagoya University, Nagoya 464-8601, Japan
We examined the effects of Rho kinase on contraction and intracellular Ca2+ concentration ([Ca2+]i) in guinea pig trachealis by measuring isometric force and the fura 2 signal [340- to 380-nm fluorescence ratio (F340/F380)]. A Rho kinase inhibitor, Y-27632 (1-1,000 µM), inhibited methacholine (MCh)-induced contraction, with a reduction in F340/F380 in a concentration-dependent manner. The values of EC50 for contraction and F340/F380 induced by 1 µM MCh with Y-27632 were 27.3 ± 5.1 and 524.1 ± 31.0 µM, respectively. With 0.1 µM MCh, the values for these parameters were decreased to 1.0 ± 0.1 and 98.2 ± 6.2 µM, respectively. Tension-F340/F380 curves for MCh indicated that Y-27632 caused an ~50% inhibition of MCh-induced contraction, without a reduction in F340/F380. These effects of Y-27632 were not inhibited by a protein kinase C inhibitor, GF-109203X. Our results indicate that inhibition of Rho kinase attenuates both Ca2+ sensitization and [Ca2+]i.
methacholine; calcium sensitivity; fura 2; protein kinase C; small G proteins
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