Perifused equine anterior pituitary cells were used to investigate the effect of cortisol on the ACTH response to pulses of corticotrophin-releasing hormone (CRH; 0·01 nmol/l) and arginine vasopressin (AVP; 100 nmol/l), given for 5 min every 30 min for 690 min and ACTH measured in 5-min fractions. At the fourth pulse of secretagogue (0 min), a constant perifusion with cortisol began (0 nmol/l (control), 100, 200, 500, 5000 and 50 000 nmol/l) and continued until the ninth pulse (150 min). For each pulse of secretagogue, the amount of ACTH (pmol) secreted in response to each pulse (ACTH response area), the highest concentration of ACTH (μg/l) measured after each pulse (peak height) and the mean ACTH in the three prepulse fractions (ACTH baseline) were determined. Data from control columns in each experiment were fitted by least squares to an exponential function to produce a mean control value for each end-point; results in all columns were expressed as a percentage of the mean control values.
The addition of cortisol had a highly significant negative effect on ACTH response area, peak height and baseline at all times from + 30 to + 240 min (columns given cortisol compared with the mean of control column values by t-test). Analysis of variance of the data showed that the higher the cortisol concentration, the quicker the ACTH response area (P = 0·0072) and peak height (P = 0·002) decreased to < 50% of mean control, and the greater the maximum percentage change (suppression) in ACTH response area (P <0·0001) and peak height (P <0·0001). The maximum percentage change (suppression) in base-line was independent of cortisol concentration.
At + 30 min after the start of cortisol perifusion, the ACTH response area in CRH columns was significantly lower than in AVP columns (P = 0·0088), and remained lower 90 min after the end of perifusion (P = 0·0084) but the maximum percentage change (suppression) was not different between secretagogues. ACTH peak height was significantly (P < 0·0268) lower in CRH than in AVP columns (from + 30 min until 180 min after the end of cortisol perifusion) and the maximum percentage change (suppression) was also greater (P = 0·0011) in CRH columns.
This study shows the effect of different concentrations of cortisol on CRH- and AVP-induced ACTH secretion by equine anterior pituitary cells, and the time-course for ACTH responses to be inhibited by, and recover from, cortisol perifusion. The highly significant inhibitory effect of cortisol on stimulated ACTH secretion was more apparent when CRH was the secretagogue than when AVP was the secretagogue. The significant inhibitory effect of cortisol on unstimulated baseline secretion of ACTH has not been described previously. These effects occur at physiological concentrations of secretagogues and cortisol. This suggests that, in vivo, circulating cortisol may have an important role in the control of ACTH secretion at pituitary level.
Journal of Endocrinology (1993) 137, 403–412
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