title: Involvement of multiple PKC isoforms in phorbol 12,13-dibutyrate-induced contraction during high K(+) depolarization in bronchial smooth muscle of mice.
abstract: In airway smooth muscle, protein kinase C (PKC) has been implicated in a number of functional responses including the regulation of contractility. However, the exact role of PKC on bronchial smooth muscle (BSM) contraction is still unclear. In the present study, to determine the role of PKC activation in the BSM contraction, the effects of phorbol 12,13-dibutyrate (PDBu, a direct PKC activator) on BSM tone were examined in the absence and presence of K(+)-induced depolarization stimulation. The force development was not evoked by treatment with 1 µM PDBu alone. However, a strong contraction was induced by PDBu during high K(+) contraction. The contraction induced by PDBu during high K(+) stimulation was significantly abolished by pretreatment with nicardipine, an L-type voltage dependent Ca(2+) channel blocker. In RT-PCR analysis, mRNAs of PKCα, β, γ, δ, ε, η and θ isoforms were detected in mouse BSM. Gö6976 (an inhibitor of PKCs α and β) and rottlerin (an inhibitor of PKCδ) significantly but partially inhibited the PDBu-induced BSM contraction during K(+) stimulation. GF109203X (an inhibitor of PKCs α, β, γ, and ε) completely inhibited the PDBu-induced contraction during K(+) stimulation. In conclusion, it is suggested that the PDBu-induced BSM contraction is dependent on an increase in cytosolic Ca(2+). Furthermore, it is possible that both cPKC and nPKC(s) participate in the PDBu-induced contraction of mouse BSM during K(+) stimulation.
authors.affiliation: Department of Pharmacology, School of Pharmacy, Hoshi University, Shinagawa-ku, Tokyo, Japan. sakai@hoshi.ac.jp
authors.forename: Hiroyasu
authors.initials: H
authors.lastname: Sakai
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Kurihara
authors.forename: Yuki
authors.initials: Y
authors.lastname: Hashimoto
authors.forename: Yoshihiko
authors.initials: Y
authors.lastname: Chiba
authors.forename: Miwa
authors.initials: M
authors.lastname: Misawa
mesh_terms: D000818:Animals; D001980:Bronchi / Q000187:drug effects* / Q000201:enzymology; D002273:Carcinogens / Q000494:pharmacology*; D007527:Isoenzymes; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D009119:Muscle Contraction / Q000502:physiology*; D009129:Muscle Tonus / Q000187:drug effects; D009130:Muscle, Smooth / Q000187:drug effects* / Q000201:enzymology; D015240:Phorbol 12,13-Dibutyrate / Q000494:pharmacology*; D011188:Potassium / Q000378:metabolism*; D011493:Protein Kinase C / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D002273:Carcinogens; D007527:Isoenzymes; D012333:RNA, Messenger; D015240:Phorbol 12,13-Dibutyrate; D011493:Protein Kinase C; D011188:Potassium