Reduced activity of acetylcholinesterase in canine tracheal smooth muscle homogenates after active immune-sensitization.
Previous investigations have suggested that immune-sensitization increases airway smooth muscle responsiveness to cholinomimetic stimulation by reducing the rate of degradation of acetylcholine. To examine the hypothesis that increased cholinomimetic responsiveness of tracheal smooth muscle (TSM) caused by immune-sensitization results from inhibition of acetylcholinesterase (AChase) activity, we developed a method for direct measurement of AChase activity in homogenates of TSM obtained from mongrel dogs actively sensitized in vivo to ragweed pollen extract (n = 7) and sham-sensitized littermate controls (n = 7). For both sensitized and control specimens, saturation of AChase was obtained at approximately 3.12 mM substrate (acetylthiocholine); however, maximal enzyme activity in homogenates of ragweed-sensitized tissues was significantly less (0.862 +/- 0.088 absorbance units/min/mg protein [AU/min/mg]) compared to control homogenates (1.590 +/- 0.129 AU/min/mg; P less than 0.001). Kinetic analysis (Eadie-Hofstee plot) indicated similar Michaelis constants (Km) for AChase from ragweed-sensitized (0.360 +/- 0.063) and control (0.336 +/- 0.062) homogenates (P = NS). The concentration of physostigmine eliciting half-maximal inhibition (Ki) of AChase activity also was similar for tissues from sensitized (-7.92 +/- 0.032 log M) and control animals (-7.86 +/- 0.012 log M; P = NS). Pretreatment with selected mediators of anaphylaxis (10(-4) M histamine, 10(-6) M serotonin, 10(-5) M prostaglandin E2, 10(-6) M prostaglandin F2 alpha, and 10(-7) M leukotriene D4) did not affect AChase activity. Our data demonstrate reduced AChase activity in homogenates of canine TSM after active immune-sensitization in vivo. This corresponds to functional augmentation of cholinomimetic contraction in actively sensitized tissues.
Mitchell
R W
RW
Department of Medicine, University of Chicago, Illinois 60637.
Kelly
E
E
Leff
A R
AR
D000110:Acetylcholinesterase / Q000276:immunology / Q000378:metabolism*; D000707:Anaphylaxis / Q000201:enzymology / Q000276:immunology; D000818:Animals; D016718:Arachidonic Acid; D001095:Arachidonic Acids / Q000378:metabolism; D002800:Cholinesterase Inhibitors / Q000494:pharmacology; D004285:Dogs; D005260:Female; D007110:Immunity, Active; D007114:Immunization; D007700:Kinetics; D008297:Male; D009119:Muscle Contraction / Q000187:drug effects; D009130:Muscle, Smooth / Q000201:enzymology* / Q000276:immunology; D010830:Physostigmine / Q000494:pharmacology; D013379:Substrate Specificity; D014132:Trachea / Q000187:drug effects / Q000201:enzymology* / Q000276:immunology
D001095:Arachidonic Acids; D002800:Cholinesterase Inhibitors; D016718:Arachidonic Acid; D010830:Physostigmine; D000110:Acetylcholinesterase