Fig. 6. Ropivacaine (Rop)–induced phosphorylation of protein kinase C (PKC) and p44/42 mitogen-activated protein kinase (p44/42 MAPK) and Rho-kinase membrane translocation in rat aortic smooth muscle. Endothelium-denuded strips were randomly challenged by different concentrations of ropivacaine for 20 min or were first pretreated with bisindolylmaleimide I (Bis I; 10−5m) ( A ), calphostin C (Calp C; 10−6m) ( B ), Y 27632 (Y; 5 × 10−6m) ( C ), or PD 098059 (PD; 10−4m) ( D ) for 15 min and then challenged with ropivacaine (3 × 10−4m) for 20 min. PKC and p44/42 MAPK phosphorylation and Rho-kinase (Rock-2) membrane translocation were detected using Western blotting with specific antibodies. Values for phosphorylation of PKC and p44/42 MAPK were expressed as percentages of the total PKC and total p44/42 MAPK, respectively. The level of Rock-2 in the membrane fraction was expressed as a percentage of the total Rock-2 ( i.e. , membrane fraction plus cytosolic fraction). * P < 0.05, P < 0.01 versus control (0 m) (n = 4). C = cytosolic fraction; M = membrane fraction; p-p44/42 MAPK = phosphorylated p44/42 mitogen-activated protein kinase; p-PKC = phosphorylated protein kinase C.
