Short Communications
165
Honn. Metab. Res. 9 (1977) 165-166
© Georg Thieme Verlag Stuttgart lack of Effect of Somatostatin on TSH Induced T3 ReJease and Platelet Function in Normal Man J. Faber, J. Gormsen, Th. Friis, C. Kirkegaard, U. Birk Lau ridsen, J. Nerup, P. Rogowski and K. Siersbaek-Nielsen Medical Department E, Frederiksberg Hospital and Coagulation Laboratory, Sundby Hospital, Copenhagen, Denmark
Since 1973 several studies have shown an inhibition by somatostatin on the release of thyroid-stimulating honnone (TSH), growth hormone, insulin, glucagon and gastrin (Anon. 1974). However, only sparse infonnation is available concerning the influence of somatostatin on the thyroid gland (Birk, Meyer, Rothenbuchner, Raptis, Loos, Adam and Pfeiffer 1976). Moreover, in 1975 conflicting results concerning the influence of somatostatin on hemostasis have been published (Besser, Paxton, Johnson, Moody, Mortimer, Hall, Gomez-Pan, Schally, Kastin and Coy 1975, Chiang, Duckworth, Beachey and Kang 1975, Koerker, Harker and Goodner 1975, Mielke, Gerich, Lorenzi, Tsalikian, Rodvien and Forsham 1975). We have studied the effect of somatostatin on TSH induced trüodothyronine (T 3) release and on platelet function in normal man.
/;:. T3 (ngl100mll 100
Figure. Increase in serum T 3 (6 T 3, mean ± SEM) following injection of bovine TSH 7.5 mU/kg in 7 healthy volunteers during an after somatostatin (e - -e ) and in 20 controls (0 - - - 0). The shaded area represent the period of infusion. Results The T 3 response to TSH stimulation was unaffected by somatostatin infusion (Figure). The biological efficacy of the somatostatin preparation used was confirmed by measuring serum insulin. In all su bjects a pronounced suppression of insulin levels was found during the somatostatin infusion.
50
The minimal concentration of ADP giving secondary platelet aggregation was 3.2 ± 2.2 pmol/ml (mean ± SD) both before and after somatostatin infusion. Further, the maximal deflection in the primary aggregation and the deflection 5 min after the addition of ADP were unchanged. The same investigaO ~~~~----r---.---, tions performed with addition of collagen also were unaffected by somato-30 0 60 120 180 240 MIN. statin. No significant difference in serotonin release after adding ADP 5 pmol/ ml was found (11.3 ± 12.2% before and 8.5 ± 10.1% after somatostatin). Neither could any difference in serotonin release be demonstrated after adding collagen 10 lJg/ml. Fibrinogen related antigens, too, were unchanged (1.05 ± 0.40 mg/I). TSH
1
Discussion
Birk et al. (1976) in 3 normal subjects found a decrease in TSH stimulated PB131j release after somato~tatin concluding that somatostatin blocks thyroid hormone synthesis or release. Our results suggest that somatostatin has no effect on TSH stimulated T 3 release in nonnal man. Our fmding of an unaffected platelet function during somatostatin infusion is in agreement with Mielke et al. (1975) but contrasts to the impaired platelet aggregation described by Besser et al. (1975). The infusion rate in these studies was comparable with the present but the infusion period was 4 to 6 hours. In animal studies using greater somatostatin doses Koerker, Harker and Goodner (1975) have shown an inhibition of platelet aggregation and Chiang et al. (1975) demonstrated that this inhibition was dose related. It is possible that the different findings are due to different preparations as the doses of somatostatin in the human studies are comparable. Acknowledgement This study was supported by Danish Hospital Foundation for Medical Research, Region of Copenhagen, Faroe Islands and Greenland. Ree.: 10 Nov. 1976
Ace.: 10 Jan. 1977
Downloaded by: University of Illinois. Copyrighted material.
Materials and Methods The subjects were 27 healthy volunteers without any signs of endocrine disorders (6 women and 21 men, aged 18-32 years). Informed consent was obtained from each subjecL Bovine TSH (Thyrotropin, Ferring) 7.5 mU/kg body weight was given as an Lv. bolus at 0 min. In 7 subjects somatostatin (a gift from Dr. J . Mulder, Ferring AB, Malmö, Sweden) 200 lJg was given as an Lv. bolus 30 minutes before the TSH injection and followed by a constant infusion of 1200 lJg until 150 minutes after the TSH injection. 20 subjects received saline only. Venous blood was drawn without compression at -30, 0, 60, 90,120, 150, 180 and 240 min. Serum T 3 (Kirkegaard, Friis and Siersbaek-Nielsen 1974) and serum insulin (Deckert, Lauridsen, Madsen and Deckert 1972) were estimated by radioimmunoassays. In 4 subjects the hemostatic function was evaluated half an hour before and 5 hours after start of infusion of somatostatin. The platelet aggregation was evaluated according to Yamazaki, Takahashi and Sano (1975). Serotonin release was estimated by the method of Gormsen (to be published) and fibrin related antigens as described by Gormsen and Clausen (1973).
166
Short Communications
References Anon.: Lancet I: 1148 (1974). - Besser, G.M., A.M. Paxton, S.A.N. Johnson, E.J. Moody, C.H. Mortimer, R. Hall, A. Gomez·Pan, A. V. Schally, A.J. Kastin, D.H. Coy: Lancet I: 1166 (1975). - Birk, J., G. Meyer, G. Rothenbuchner, S. Raptis, U. Loos, W.E. Adam, E.F. Pfeiffer: Acta Endocrinol. Suppl. 204: 50 (1976). - Chiang, T.M., W.c. Duckworth, E.H. Beachey, A.H. Kang: Endocrinology 97: 753 (1975). - Deckert, T., U.B. Lauridsen, S.N. Madsen, M. Deckert: Horm.Metab.Res. 4: 229 (1972). - Gormsen, J., N. T. Clausen: Ugeskr.Laeger 135: 1682 (1973). - Kirkegaard, c., Th. Friis, fC Siersbaek·Nie/· sen: Acta EndocrinoL 77: 71 (1974). - Koerker, D.J., L.A. Harker, c.J. Goodner: N.EngI.J.Med. 293: 476 (1975). Mie/ke, C.H., J.E. Gerich, M. Lorenzi, E. Tsalikian, R. Rodvien, P.H. Forsham: N.Engl.J.Med. 293: 480 (1975). - Yamazaki, H., T. Takahashi, T. Sano: Thrombos.Diathes.Haemorrh. 34: 94 (1975). Requests for reprints should be addressed to: Dr. J. Faber, Medical Department E, Frederiksberg Hospital, DK-2000 Copenhagen F (Denmark)
Horm. Metab. Res. 9 (1977) 166
© Georg Thieme Verlag Stuttgart Cushing's Syndrome and Serum Calcium-Albumin Interaction
Chronic glucocorticoid excess in man can be associated with several alterations in Ca metabolism (Wajchenberg, Pereira, Kieffer and Ursic 1969). Collins, Ga"et and Johnston (1962) speculated that this hormone might induce a relative increase in the diffusible Ca fraction. In this case, an alteration in the normal affinity of Ca to serum albumin might be present in Cushing's syndrome. This possibility was investigated in this paper. Material and Methods 7 patients with Cushing's syndrome (CSP) (1 adenoma, 2 hyperplasias and 4 adrenal carcinomas), 4 males and 3 females, ages 3-35 yrs and 8 normal adult subjects (NS) (2 males and 6 females) were studied. Blood was collected aerobicalJy without stasis in the fasting state. 3 aliquots with decreasing pHs were prepared from each serum sarnple by intermittent gassing with O2 + CO 2 (5%) (3-6 min). Ca++ (Orion flow-through, 98-20), pH (IL pH BJood-Gas Analyser 313), albumin (Ness, Dickerson and Pastewka 1965) and total proteins (Weichse/baum 1946) were determined in serum. A multiple regression analysis was performed and the best linear equation correlating serum Ca++ (dependent variable) and serum pH and albumin was derived for both NS and CSP. Statistical analysis was performed by the standard error (and respective t values) of the coefTicients for pH and albumin. Student's t test was used to evaluate differences of 2 means (Dixon and Massey Jr. 1957). The rationale of this method of studying serum Ca-albumin interaction was presented in a previous publication (Leme, Wachslicht and Wajchenberg 1976). Results Mean serum albumins ± 1 SEM were 4.31 ± 0.12 for the NS and 3.64 ± 0.12 g/ 100 ml for the CSP (p < 0.05). Mean serum globulins ± 1 SEM were 2.72 ± 0.14 for the NS and 2.65 ± 0.12 g/ 100 ml for the CSP (p 0.05). Multiple regression equations were: NS:Ca++ = 9.6-1.09 pH - 0.14 Alb. CSP:Ca++ = 6.3-0.71 pH - 0.03 Alb. pH coefficients were significant (p < 0.001) for both NS and CSP showing that "in vitro" pH reductions increase Ca++ in serum. Albumin coefficients were not statistically significant for both groups indicating that serum Ca++ is not influenced by the previous albumin level of the subjects. pH coefficients were significantly greater for the NS in relation to CSP (p < 0.001). To verify if these differences were exclusively albumin-related, selected data on Ca++ (dependent variable) and pH from 2 NS and 2 CSP with similar albumin levels were tested by regression analysis. The equations were: NS:r = -0.99, p
Discussion Serum from CSP increases less than normal its Ca++ with pH reductions. This different behaviour eannot be entirely attributed to the lower serum albumin of the CSP, as it occurred also at similar albumin levels. The results suggest that in Cushing's syndrome there may occur an increased binding affinity of Ca to serum albumin. Referenees
Wajchenberg, B.L., V.G. Pereira, J. Kieffer, S. Ursic: Acta Endoerinol. 61: 173 (1969). - Collins, E.J., E.R. Ga"et, R.L. Johnston: Metabolism 11: 716 (1962). - Ness, A.T., H.C. Dickerson, J.V. Pastewka: Clin.Chim.Aeta 12: 532 (1965). Weichse/baum, T.E.: Amer.J.Clin.Pathol. 16: 40 (1946). - Dixon, W.J., F.J. Massey Jr.: Introduction to statistical analysis, 2nd,Ed. McGraw-Hill (1957). - Leme, C.E., H. Wachslicht, B.L. Wajchenberg: Horm.Metab.Res. 8: 326 (1976). Requests for reprints should be addressed to: C.E. Lerne, Hosp. Clfnicas, Säo Paulo, Brazil
Rec.: 25 Aug. 1976
Ace.: 10 Jan. 1977
Downloaded by: University of Illinois. Copyrighted material.
C.E. Lerne, B.L Wajchenberg, P.R. Lerne and F.P. Cesar Diabetes Unit, Hospital das CI(nicas, säo Paulo, Brazil
165
Honn. Metab. Res. 9 (1977) 165-166
© Georg Thieme Verlag Stuttgart lack of Effect of Somatostatin on TSH Induced T3 ReJease and Platelet Function in Normal Man J. Faber, J. Gormsen, Th. Friis, C. Kirkegaard, U. Birk Lau ridsen, J. Nerup, P. Rogowski and K. Siersbaek-Nielsen Medical Department E, Frederiksberg Hospital and Coagulation Laboratory, Sundby Hospital, Copenhagen, Denmark
Since 1973 several studies have shown an inhibition by somatostatin on the release of thyroid-stimulating honnone (TSH), growth hormone, insulin, glucagon and gastrin (Anon. 1974). However, only sparse infonnation is available concerning the influence of somatostatin on the thyroid gland (Birk, Meyer, Rothenbuchner, Raptis, Loos, Adam and Pfeiffer 1976). Moreover, in 1975 conflicting results concerning the influence of somatostatin on hemostasis have been published (Besser, Paxton, Johnson, Moody, Mortimer, Hall, Gomez-Pan, Schally, Kastin and Coy 1975, Chiang, Duckworth, Beachey and Kang 1975, Koerker, Harker and Goodner 1975, Mielke, Gerich, Lorenzi, Tsalikian, Rodvien and Forsham 1975). We have studied the effect of somatostatin on TSH induced trüodothyronine (T 3) release and on platelet function in normal man.
/;:. T3 (ngl100mll 100
Figure. Increase in serum T 3 (6 T 3, mean ± SEM) following injection of bovine TSH 7.5 mU/kg in 7 healthy volunteers during an after somatostatin (e - -e ) and in 20 controls (0 - - - 0). The shaded area represent the period of infusion. Results The T 3 response to TSH stimulation was unaffected by somatostatin infusion (Figure). The biological efficacy of the somatostatin preparation used was confirmed by measuring serum insulin. In all su bjects a pronounced suppression of insulin levels was found during the somatostatin infusion.
50
The minimal concentration of ADP giving secondary platelet aggregation was 3.2 ± 2.2 pmol/ml (mean ± SD) both before and after somatostatin infusion. Further, the maximal deflection in the primary aggregation and the deflection 5 min after the addition of ADP were unchanged. The same investigaO ~~~~----r---.---, tions performed with addition of collagen also were unaffected by somato-30 0 60 120 180 240 MIN. statin. No significant difference in serotonin release after adding ADP 5 pmol/ ml was found (11.3 ± 12.2% before and 8.5 ± 10.1% after somatostatin). Neither could any difference in serotonin release be demonstrated after adding collagen 10 lJg/ml. Fibrinogen related antigens, too, were unchanged (1.05 ± 0.40 mg/I). TSH
1
Discussion
Birk et al. (1976) in 3 normal subjects found a decrease in TSH stimulated PB131j release after somato~tatin concluding that somatostatin blocks thyroid hormone synthesis or release. Our results suggest that somatostatin has no effect on TSH stimulated T 3 release in nonnal man. Our fmding of an unaffected platelet function during somatostatin infusion is in agreement with Mielke et al. (1975) but contrasts to the impaired platelet aggregation described by Besser et al. (1975). The infusion rate in these studies was comparable with the present but the infusion period was 4 to 6 hours. In animal studies using greater somatostatin doses Koerker, Harker and Goodner (1975) have shown an inhibition of platelet aggregation and Chiang et al. (1975) demonstrated that this inhibition was dose related. It is possible that the different findings are due to different preparations as the doses of somatostatin in the human studies are comparable. Acknowledgement This study was supported by Danish Hospital Foundation for Medical Research, Region of Copenhagen, Faroe Islands and Greenland. Ree.: 10 Nov. 1976
Ace.: 10 Jan. 1977
Downloaded by: University of Illinois. Copyrighted material.
Materials and Methods The subjects were 27 healthy volunteers without any signs of endocrine disorders (6 women and 21 men, aged 18-32 years). Informed consent was obtained from each subjecL Bovine TSH (Thyrotropin, Ferring) 7.5 mU/kg body weight was given as an Lv. bolus at 0 min. In 7 subjects somatostatin (a gift from Dr. J . Mulder, Ferring AB, Malmö, Sweden) 200 lJg was given as an Lv. bolus 30 minutes before the TSH injection and followed by a constant infusion of 1200 lJg until 150 minutes after the TSH injection. 20 subjects received saline only. Venous blood was drawn without compression at -30, 0, 60, 90,120, 150, 180 and 240 min. Serum T 3 (Kirkegaard, Friis and Siersbaek-Nielsen 1974) and serum insulin (Deckert, Lauridsen, Madsen and Deckert 1972) were estimated by radioimmunoassays. In 4 subjects the hemostatic function was evaluated half an hour before and 5 hours after start of infusion of somatostatin. The platelet aggregation was evaluated according to Yamazaki, Takahashi and Sano (1975). Serotonin release was estimated by the method of Gormsen (to be published) and fibrin related antigens as described by Gormsen and Clausen (1973).
166
Short Communications
References Anon.: Lancet I: 1148 (1974). - Besser, G.M., A.M. Paxton, S.A.N. Johnson, E.J. Moody, C.H. Mortimer, R. Hall, A. Gomez·Pan, A. V. Schally, A.J. Kastin, D.H. Coy: Lancet I: 1166 (1975). - Birk, J., G. Meyer, G. Rothenbuchner, S. Raptis, U. Loos, W.E. Adam, E.F. Pfeiffer: Acta Endocrinol. Suppl. 204: 50 (1976). - Chiang, T.M., W.c. Duckworth, E.H. Beachey, A.H. Kang: Endocrinology 97: 753 (1975). - Deckert, T., U.B. Lauridsen, S.N. Madsen, M. Deckert: Horm.Metab.Res. 4: 229 (1972). - Gormsen, J., N. T. Clausen: Ugeskr.Laeger 135: 1682 (1973). - Kirkegaard, c., Th. Friis, fC Siersbaek·Nie/· sen: Acta EndocrinoL 77: 71 (1974). - Koerker, D.J., L.A. Harker, c.J. Goodner: N.EngI.J.Med. 293: 476 (1975). Mie/ke, C.H., J.E. Gerich, M. Lorenzi, E. Tsalikian, R. Rodvien, P.H. Forsham: N.Engl.J.Med. 293: 480 (1975). - Yamazaki, H., T. Takahashi, T. Sano: Thrombos.Diathes.Haemorrh. 34: 94 (1975). Requests for reprints should be addressed to: Dr. J. Faber, Medical Department E, Frederiksberg Hospital, DK-2000 Copenhagen F (Denmark)
Horm. Metab. Res. 9 (1977) 166
© Georg Thieme Verlag Stuttgart Cushing's Syndrome and Serum Calcium-Albumin Interaction
Chronic glucocorticoid excess in man can be associated with several alterations in Ca metabolism (Wajchenberg, Pereira, Kieffer and Ursic 1969). Collins, Ga"et and Johnston (1962) speculated that this hormone might induce a relative increase in the diffusible Ca fraction. In this case, an alteration in the normal affinity of Ca to serum albumin might be present in Cushing's syndrome. This possibility was investigated in this paper. Material and Methods 7 patients with Cushing's syndrome (CSP) (1 adenoma, 2 hyperplasias and 4 adrenal carcinomas), 4 males and 3 females, ages 3-35 yrs and 8 normal adult subjects (NS) (2 males and 6 females) were studied. Blood was collected aerobicalJy without stasis in the fasting state. 3 aliquots with decreasing pHs were prepared from each serum sarnple by intermittent gassing with O2 + CO 2 (5%) (3-6 min). Ca++ (Orion flow-through, 98-20), pH (IL pH BJood-Gas Analyser 313), albumin (Ness, Dickerson and Pastewka 1965) and total proteins (Weichse/baum 1946) were determined in serum. A multiple regression analysis was performed and the best linear equation correlating serum Ca++ (dependent variable) and serum pH and albumin was derived for both NS and CSP. Statistical analysis was performed by the standard error (and respective t values) of the coefTicients for pH and albumin. Student's t test was used to evaluate differences of 2 means (Dixon and Massey Jr. 1957). The rationale of this method of studying serum Ca-albumin interaction was presented in a previous publication (Leme, Wachslicht and Wajchenberg 1976). Results Mean serum albumins ± 1 SEM were 4.31 ± 0.12 for the NS and 3.64 ± 0.12 g/ 100 ml for the CSP (p < 0.05). Mean serum globulins ± 1 SEM were 2.72 ± 0.14 for the NS and 2.65 ± 0.12 g/ 100 ml for the CSP (p 0.05). Multiple regression equations were: NS:Ca++ = 9.6-1.09 pH - 0.14 Alb. CSP:Ca++ = 6.3-0.71 pH - 0.03 Alb. pH coefficients were significant (p < 0.001) for both NS and CSP showing that "in vitro" pH reductions increase Ca++ in serum. Albumin coefficients were not statistically significant for both groups indicating that serum Ca++ is not influenced by the previous albumin level of the subjects. pH coefficients were significantly greater for the NS in relation to CSP (p < 0.001). To verify if these differences were exclusively albumin-related, selected data on Ca++ (dependent variable) and pH from 2 NS and 2 CSP with similar albumin levels were tested by regression analysis. The equations were: NS:r = -0.99, p
Discussion Serum from CSP increases less than normal its Ca++ with pH reductions. This different behaviour eannot be entirely attributed to the lower serum albumin of the CSP, as it occurred also at similar albumin levels. The results suggest that in Cushing's syndrome there may occur an increased binding affinity of Ca to serum albumin. Referenees
Wajchenberg, B.L., V.G. Pereira, J. Kieffer, S. Ursic: Acta Endoerinol. 61: 173 (1969). - Collins, E.J., E.R. Ga"et, R.L. Johnston: Metabolism 11: 716 (1962). - Ness, A.T., H.C. Dickerson, J.V. Pastewka: Clin.Chim.Aeta 12: 532 (1965). Weichse/baum, T.E.: Amer.J.Clin.Pathol. 16: 40 (1946). - Dixon, W.J., F.J. Massey Jr.: Introduction to statistical analysis, 2nd,Ed. McGraw-Hill (1957). - Leme, C.E., H. Wachslicht, B.L. Wajchenberg: Horm.Metab.Res. 8: 326 (1976). Requests for reprints should be addressed to: C.E. Lerne, Hosp. Clfnicas, Säo Paulo, Brazil
Rec.: 25 Aug. 1976
Ace.: 10 Jan. 1977
Downloaded by: University of Illinois. Copyrighted material.
C.E. Lerne, B.L Wajchenberg, P.R. Lerne and F.P. Cesar Diabetes Unit, Hospital das CI(nicas, säo Paulo, Brazil
