The effect of pertussis toxin on beta-adrenoceptor responses in isolated cardiac myocytes from noradrenaline-treated guinea-pigs and patients with cardiac failure.
1. A decreased responsiveness to the positive inotropic effects of beta-adrenoceptor agonists is a characteristic of human heart failure. We have investigated the involvement of inhibitory guanine nucleotide binding proteins (G-proteins) in this process using pertussis toxin treatment of isolated cardiac myocytes. 2. Myocytes isolated from failing human myocardium had a reduced maximum contractile response to isoprenaline relative to that for maximally stimulating concentrations of calcium, giving an isoprenaline/calcium ratio of 0.71 +/- 0.06 (n = 7 patients). This was significantly lower than in myocytes from non-failing myocardium, where the isoprenaline/calcium ratio was 1.16 +/- 0.07 (n = 6, P less than 0.001). Responses to high calcium were unchanged. 3. Myocytes were treated with pertussis toxin for 3-5 h at 35 degrees C. Successful inactivation of inhibitory G-proteins by pertussis toxin treatment was indicated by a loss of responsiveness to 10 microM adenosine (in the presence of submaximal isoprenaline). 4. Following pertussis toxin treatment of the myocytes from failing human heart the isoprenaline/calcium ratio increased to 1.43 +/- 0.27 (n = 7, P less than 0.05). Pertussis toxin treatment had no effect upon the maximum calcium contraction. The isoprenaline/calcium ratio in myocytes from non-failing human ventricle was not affected by the toxin treatment (n = 3). These observations support the hypothesis that increased inhibitory G-protein levels or activities contribute to beta-adrenoceptor desensitization in human heart failure. 5. beta-Adrenoceptor desensitization in human heart failure is thought to be secondary to raised noradrenaline levels in these patients. Experiments were repeated on myocytes isolated from hearts of guinea-pigs which had been chronically infused with noradrenaline. The isoprenaline/calcium ratio of these myocytes was reduced below that of myocytes from sham-operated animals (0.65 0.04, n = 6 compared with 0.88 +/- 0.02, n = 7, P<0.001).6. Pertussis toxin treatment (2 h at 35 degrees C) increased the isoprenaline/calcium ratio to 1.02 0.02 (n = 6,P<0.01) in myocytes from noradrenaline-treated guinea-pigs. This effect of pertussis toxin treatment was not seen in myocytes from sham-operated guinea-pig hearts.7. Incubation at 35 degrees C for similar periods in the absence of pertussis toxin also restored the isoprenaline/calcium ratio towards normal in the myocytes from both failing human and noradrenaline-treated guinea-pig hearts, although the effect was significantly smaller than that produced by the toxin. Myocytes kept at room temperature (22 degrees C) showed no such evidence of resensitization over periods up to 6h.8. These observations are consistent with the hypothesis that raised catecholamine levels result in increased inhibitory G-protein levels and functional P-adrenoceptor desensitization in heart failure.
Brown
L A
LA
Department of Cardiac Medicine, National Heart & Lung Institute, London.
Harding
S E
SE
D000328:Adult; D000704:Analysis of Variance; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002303:Cardiac Output, Low / Q000503:physiopathology*; D002648:Child; D005260:Female; D019204:GTP-Binding Proteins / Q000378:metabolism; D006168:Guinea Pigs; D006321:Heart / Q000187:drug effects*; D006801:Humans; D007545:Isoproterenol / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D009200:Myocardial Contraction / Q000187:drug effects*; D009206:Myocardium / Q000166:cytology; D009638:Norepinephrine / Q000494:pharmacology; D037342:Pertussis Toxin; D011943:Receptors, Adrenergic, beta / Q000187:drug effects*; D010566:Virulence Factors, Bordetella / Q000494:pharmacology*
D011943:Receptors, Adrenergic, beta; D010566:Virulence Factors, Bordetella; D037342:Pertussis Toxin; D019204:GTP-Binding Proteins; D007545:Isoproterenol; D002118:Calcium; D009638:Norepinephrine