Role of Rho kinase in endothelin-1-induced phosphorylation of CPI-17 in rat bronchial smooth muscle

It has been reported that CPI-17 (protein kinase C (PKC)-potentiated inhibitory protein for heterotrimeric myosin light chain phosphatase (MLCP) of 17 kDa) was phosphorylated by excitatory agonists in smooth muscle contraction. However, endothelin-1 (ET-1)-mediated regulation of CPI-17 in bronchial smooth muscle has not been documented. We therefore investigated whether phosphorylation of CPI-17 is induced by ET-1 in rat bronchial smooth muscle. Moreover, the role of Rho kinase (ROCK; Rho-associated coiled-coil forming protein kinase) is investigated in phosphorylation of CPI-17 induced by ET-1 in rat bronchial smooth muscle. The ET-1-induced contraction was attenuated by Y-27632 (10−6 M), a ROCK inhibitor. ET-1 induced a phosphorylation of CPI-17 with a phosphorylation of myosin light chain (MLC); those phosphorylation responses were significantly inhibited by Y-27632 (10−6 M). These findings suggest that the activation of ROCK is involved in force development and CPI-17 phosphorylation induced by ET-1 stimulation in rat bronchial smooth muscle. Thus, RhoA/ROCK/CPI-17 pathway is considered to play an important role in the ET-1-induced Ca2+ sensitisation of bronchial smooth muscle contraction.