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Calcium sensitization of smooth muscle mediated by a Rho-associated protein kinase in hypertension

Abstract

Abnormal smooth-muscle contractility may be a major cause of disease states such as hypertension, and a smooth-muscle relaxant that modulates this process would be useful therapeutically. Smooth-muscle contraction is regulated by the cytosolic Ca2+ concentration and by the Ca2+ sensitivity of myofilaments1: the former activates myosin light-chain kinase and the latter is achieved partly by inhibition of myosin phosphatase1,2,3. The small GTPase Rho and its target, Rho-associated kinase, participate in this latter mechanism in vitro4,5,6, but their participation has not been demonstrated in intact muscles. Here we show that a pyridine derivative, Y-27632, selectively inhibits smooth-muscle contraction by inhibiting Ca2+ sensitization. We identified the Y-27632 target as a Rho-associated protein kinase, p160ROCK7. Y-27632 consistently suppresses Rho-induced, p160ROCK-mediated formation of stress fibres in cultured cells and dramatically corrects hypertension in several hypertensive rat models. Our findings indicate that p160ROCK-mediated Ca2+ sensitization is involved in the pathophysiology of hypertension and suggest that compounds that inhibit this process might be useful therapeutically.

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Figure 1: Effects of Y-27632 on smooth muscle contraction.
Figure 2: Identification of p160ROCK as a target for Y-27632.
Figure 3: Inhibition by Y-27632 of the Rho-induced formation of focal adhesions and stress fibres, but not Rac-induced membrane ruffling.
Figure 4: Effects of Y-27632 on blood pressure.

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Acknowledgements

We thank S. Ohno and R. Cerione for recombinant PKCε and PAK cDNA, respectively, H. Karaki for helpful suggestions, H. Bito for critical reading of the manuscript, and K. Okuyama for secretarial assistance. Supported in part by a grant-in-aid for specially promoted research from the Ministry of Education, Science and Culture of Japan, and by an HFSP grant.

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Correspondence to Masayoshi Uehata or Shuh Narumiya.

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Uehata, M., Ishizaki, T., Satoh, H. et al. Calcium sensitization of smooth muscle mediated by a Rho-associated protein kinase in hypertension. Nature 389, 990–994 (1997). https://doi.org/10.1038/40187

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