The direct effects of heparin and protamine on canine tracheal smooth muscle tone.
Heparin and protamine are used for cardiopulmonary bypass in cardiac surgery; however, the direct effects and mechanisms of these drugs on airway smooth muscle tone are still not fully known. We investigated the in vitro effects of these drugs on canine tracheal smooth muscle by measuring the muscle tension and intracellular Ca2+ concentration ([Ca2+]i) and by measuring inward Ca2+ currents (I(Ca)) through voltage-dependent Ca2+ channels. [Ca2+]i was monitored by the 500-nm light emission ratio of preloaded Ca2+ indicator fura-2. Isometric tension was measured simultaneously. Whole-cell patch clamp recording techniques were used to investigate the effects of the drugs on I(Ca) in freshly dispersed smooth muscle cells. Heparin (0.12-120 U/mL), protamine (0.15-150 U/mL), or heparin-protamine complex (4:5 U/U) was introduced into a bath solution. Protamine and heparin-protamine complex dose-dependently inhibited both carbachol-induced contraction of the muscle and increase in [Ca2+]i. These drugs also decreased the I(Ca) of the muscle cells and shifted the inactivation curve to a more negative potential. Heparin itself had a slight enhancing effect on carbachol-induced muscle contraction without changing [Ca2+]i. Protamine and heparin-protamine complex can decrease the agonist-induced increase in [Ca2+]i by the inhibition of voltage-dependent Ca2+ channels both in the activated and inactivated states.


CONCLUSIONS
Protamine and heparin-protamine complex inhibited carbachol-induced canine tracheal smooth muscle contraction by inhibiting the increase in intracellular concentration of free Ca2+. These drugs can decrease the agonist-induced increase in intracellular Ca2+ by the inhibition of voltage-dependent Ca2+ channels in both the activated and inactivated states.
Yamakage
M
M
Department of Anesthesiology, Sapporo Medical University School of Medicine, Hokkaido, Japan. yamakage@sapmed.ac.jp
Matsuzaki
T
T
Tsujiguchi
N
N
Mori
T
T
Namiki
A
A
D000818:Animals; D000925:Anticoagulants / Q000494:pharmacology*; D002118:Calcium / Q000378:metabolism; D015220:Calcium Channels / Q000187:drug effects / Q000502:physiology; D002217:Carbachol / Q000494:pharmacology; D004285:Dogs; D006493:Heparin / Q000494:pharmacology*; D006494:Heparin Antagonists / Q000494:pharmacology*; D066298:In Vitro Techniques; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D009129:Muscle Tonus / Q000187:drug effects; D009130:Muscle, Smooth / Q000187:drug effects*; D018408:Patch-Clamp Techniques; D011479:Protamines / Q000494:pharmacology*; D014132:Trachea / Q000187:drug effects*
D000925:Anticoagulants; D015220:Calcium Channels; D006494:Heparin Antagonists; D011479:Protamines; D002217:Carbachol; D006493:Heparin; D002118:Calcium