Rho-kinase and contractile apparatus proteins in murine airway hyperresponsiveness.
Airway hyperresponsiveness (AHR) is a hallmark of bronchial asthma. Increased expression of smooth muscle contractile proteins or increased responsiveness of the contractile apparatus due to RhoA/Rho-kinase activation may contribute to AHR. BALB/c mice developed AHR following systemic sensitization by intraperitoneal injections of 20 microg ovalbumin (OVA) in presence of 2mg Al(OH)(3) on days 1 and 14, and airway challenge by 1% OVA-inhalation for 20 min each on days 28, 29 and 30. As assessed by Western blot, protein expression of RhoA, MLC (myosin light chain) and smMLCK (smooth muscle myosin light chain kinase) was increased in lungs of OVA/OVA-animals with AHR, as well as in lungs of OVA-sensitized and sham-challenged animals (OVA/PBS) without AHR, compared with lungs of PBS/PBS-animals. Pretreatment with the specific Rho-kinase inhibitor Y-27632 reduced MLC-phosphorylation and AHR. Contribution of Rho-kinase to bronchoconstriction was increased in lungs of OVA/OVA-animals compared with OVA/PBS- and PBS/PBS-animals, respectively. Furthermore, bronchoconstriction following MCh stimulation was significantly reduced after Y-27632 application. In conclusion, systemic allergen-sensitization increased pulmonary expression of proteins involved in smooth muscle contraction, which may contribute to development of AHR. However, this observation was independent from local allergen challenge, suggesting that additional cofactors may be required for the activation of Rho-kinase and thereby the induction of AHR. Rho-kinase may play an important role in murine AHR, and the bronchodilating action of Rho-kinase inhibition may offer a new therapeutic perspective in obstructive airway disease.
Witzenrath
Martin
M
Department of Internal Medicine/Infectious Diseases and Respiratory Medicine, Charité-Universitätsmedizin Berlin, Berlin, Germany. martin.witzenrath@charite.de
Ahrens
Birgit
B
Schmeck
Bernd
B
Kube
Stefanie M
SM
Hippenstiel
Stefan
S
Rosseau
Simone
S
Hamelmann
Eckard
E
Suttorp
Norbert
N
Schütte
Hartwig
H
D000577:Amides / Q000494:pharmacology; D000818:Animals; D001249:Asthma / Q000139:chemically induced / Q000188:drug therapy / Q000201:enzymology*; D015153:Blotting, Western; D016535:Bronchial Hyperreactivity / Q000139:chemically induced / Q000188:drug therapy / Q000201:enzymology*; D016084:Bronchoconstriction / Q000187:drug effects / Q000502:physiology*; D004195:Disease Models, Animal; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D008168:Lung / Q000187:drug effects / Q000201:enzymology; D051379:Mice; D008807:Mice, Inbred BALB C; D009130:Muscle, Smooth / Q000187:drug effects / Q000201:enzymology; D018994:Myosin Light Chains / Q000187:drug effects / Q000378:metabolism*; D009219:Myosin-Light-Chain Kinase / Q000187:drug effects / Q000378:metabolism*; D010047:Ovalbumin / Q000276:immunology; D010477:Perfusion; D010766:Phosphorylation; D011725:Pyridines / Q000494:pharmacology; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*
D000577:Amides; D004791:Enzyme Inhibitors; D018994:Myosin Light Chains; D011725:Pyridines; C108830:Y 27632; D010047:Ovalbumin; D054460:rho-Associated Kinases; D009219:Myosin-Light-Chain Kinase