Rationale: Augmented bronchial smooth muscle (BSM) contraction is one of the causes of bronchial hyperresponsiveness. The protein RhoA and its downstream pathways have now been proposed as a new target for asthma therapy. MicroRNAs (miRNAs) play important roles in normal and diseased cell functions, and a contribution of miR-133 to RhoA expression has been suggested in cardiomyocytes.
Objectives: To make clear the mechanism(s) of up-regulation of RhoA observed in the BSMs of experimental asthma, the role of miR-133a in RhoA expression was tested.
Methods: Total proteins and RNAs (containing miRNAs) were extracted from cultured human BSM cells (hBSMCs) that were treated with antagomirs and/or IL-13, and bronchial tissues of BALB/c mice that were sensitized and repeatedly challenged with ovalbumin. RhoA protein and miR-133a were detected by immunoblotting and quantified real-time reverse transcriptase-polymerase chain reaction, respectively.
Measurements and main results: In hBSMCs, an up-regulation of RhoA was observed when the function of endogenous miR-133a was inhibited by its antagomir. Treatment of hBSMCs with IL-13 caused an up-regulation of RhoA and a down-regulation of miR-133a. In bronchial tissues of the repeatedly ovalbumin-challenged mice, a significant increase in RhoA was observed. Interestingly, the level of miR-133a was significantly decreased in BSMs of the challenged mice.
Conclusions: These findings suggest that RhoA expression is negatively regulated by miR-133a in BSMs. IL-13 might, at least in part, contribute to the reduction of miR-133a.