Alteration in expressions of RhoA and Rho-kinases during pregnancy in rats: their roles in uterine contractions and onset of labour.

Activation of RhoA and Rho-associated kinases (ROCKs) is known to play a pivotal role in the regulation of smooth muscle contraction via phosphorylation of myosin-light chain and myosin phosphatase. There are few data on the RhoA and ROCKs expression levels in rat uteri. Therefore, our aim was to investigate the mRNA and protein concentration of RhoA and ROCKs in rat uterus during pregnancy, during parturition and post-partum using real time PCR and Western blot analysis. The other purpose was to evaluate the effects of the ROCK (Y-27632, fasudil and RKI 1441) and RhoA inhibitors (simvastatin) on uterine contractility in isolated organ bath experiments. The mRNA and protein levels of RhoA decreased on the 5th day of pregnancy to day 22, then a sharp increase was detected at term. The mRNA and protein concentration of ROCKs was down-regulated in the early stage of pregnancy, while it sharply increased during parturition. The RhoA-inhibitor simvastatin relaxed the uterus contractions, although its inhibitory effects were not followed by the alteration of RhoA. The strongest inhibitory effect of non-selective ROCK inhibitor fasudil was found on non-pregnant uterus, while it elicited milder relaxation on day 22, during parturition and postpartum day 1. The maximum relaxing effects of Y-27632 and RKI 1441 were altered in a proportional way with the target protein expressions. The RhoA/ROCK signalling pathway might be a potential target for the development of new tocolytic agents; however, high specificity to RhoA, ROCK I or ROCK II seems to be fundamental to the high efficacy of uterine relaxation.