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Human recombinant growth hormone with and without salmon calcitonin in the treatment of postmenopausal osteoporosis
137 HUMAN RECOMBINANTGROWTH HORNONE WITH AND WITHOUT SALWON
CALCIfONIN IN THE TREATWENT OF POSTNENOPAUSAL OSTEOPOROSIS. C.Cepollaro, S.Gonnelli, M.Montomoli, A.Camporeale, D.Acrnusdei, F.Zacchei, M.S.Campagna, C.Gennari. Instituteof Internal Medicine,Universityof Siena, Italy. Some studies,performedwith extractivegrowth hormone (GH), suggested its use as stimulatorof bone formation in the treatment of postmenopausalosteoporosis (PMO). the efficacy of GH therapy in PM0 is still However, debated. Now a DNA recombinant human GH (rhGH) is available for clinical use. The present single blind study has been performed in order to compare the effects of rhGH given alone or in combination with salmon calcitonin (sCT) on bone turnover and bone mass in patients with PMO. 30 women (age 60.824 SD) with established PM0 were divided into 3 groups of 10 and randomly assigned to 3 treatment sequences:1) rhGH (12 Iv/d) S.C. for 7 days, followedby sCT (50 IU/d) S.C. for 21 days, and by 61-days free; 2) rhGH for 7 days, followed by placebo (Pl) for 21 days, and by Cl-days free; 3) Pl for 7 days, followedby sCT for 21 days, and by 61-days free. Each cycle was repeated4 times. At days 0, 8, 29 and 90 of each cycle, serum osteocalcin (BGP) and urinary excretion of Ca (CaU) and hydroryproline (IlOP were measured. QMD of thn lumbar sDine. femoral shaft, and distal radius was measurod,by IX%, defore and after 6 and 12 months. No significantchanges in EMD were observed wlthin and among thn 3 groups. With regard to the indaxos of bane turnover,a constant and significant increase2n BGP, MOP und Call wao detuctod during tha rhGl1 periods. HOP, as expected, significanllydecrescd during sC!Tperiods. Our study suggests that rhGtl acts as an activator of bone turnover rather than as o stimulatorof bone formation.
139 ACUTE EFFECT OF SHELL FORMATION ON 1,25 (OH),D, AND OSTEOCALCIN BLOOD LEVELS AND ON INTESTINAL AND UTERINE CALSINDIN mRNA IN HENS. X&&s. K. Allewatzt. I&W*, v-; INRA, Nouzilly, F37380; ‘LEGENDO, Leuven, S3oo0, ’ Cambridge, UK. Tha time-course of the changes in ionized calcium, 1,25(0H),D,, and osteocalcln blood levels and in the intestinal and uterine culblndin mRNA were studied In hens following suppression and resumption ol shell formallon and throughout the leylng cycle in hens laying hard-shelled or shell-lees eggs, In hens laying hard-shelled eggs circulallng l,25(OHJr& and osleocalcin, and uterinecalbindln mRNA were higher during the period of shell secretion but no differences were observed for intestinal calblndin mRNA. Plasma 1,25(OH)&, and osteocalcin were decreased 3h and 6h after suppression of shell formation. Uterine calbindin mRNA was depressed to 54 % of its initial value lh after egg expulsion and remained al a WY low level when eoa exoulslon was continued for 8 days. lntesllnal calblndin mRNA level was n6l affectad by suppression of shell formation. Calbindln levels decreased In both tissues only when premature ewpulslons of the egg were carried out for several da&When egg shell formallon resumed in hens previously laying sheRI-less eggs, the concenlratlon of 1,25(OH),4, osteocalcin and of inteslinal calbindin mANA increased toward Ihe end of shell formation but uterine calblndln and its mRNA were enhanced , earlier, al the onset on shell lormation. These Increases were maintalned at the uterine level in contrast lo the intestine &en hens were parathyroideclomised before shell resumption and when, consequently 1,25(0H),DS decreased. Feeding hens a tow calcium diet caused a large Increase in plasma 1,25(OH),DJ and osleocalcin. Ionized calcium levels tended to show reciprocal changes lo 1,25(0H),D,. However in hypercalcemic hens laying shelt-less eggs and fed a 3.5% calcium diet, the plasma 1,25(OH),D, was al a high level after ovulationand diminishedthereafter. In conctuslon, this last observation suggest that In hens, 1,25(OH),D, production Is regulated, in addilion lo Ihe stimulalion Invoking decreased C&I” and increased PTH, by anolher possibly endocrine factor linked lo ovulation. Acute changes in plasma 1,25(OH),D, and osleocalcin were parallel suggesting a relationship between these oaramelers. In contrast intestinalmRNA was stable posdbly as a consequ&ce of the transepilhelial calcium transport. Large daily increase in utetine calbindinmRNA occured irrespectively Of 1,25(OHJ,D, ievels and coincided with uterine calcium transpotl indicating that its regulation involves a calcium flux dependent factor.
138 FFECTS OF tllGW DOSE CALCITRIOL
ON RAT BONE
Scharing Resaarch, Allgemaine Pharmakologie. Berlin. FRG. We s&died time dependent changes in bone histology and re5paCtiVe biochemical markers of bone turnover during high dOSacalcltriol treatment (CAL) in adult (b.w.: 200 - 2409) female rats. 3 groups of weight matched animals (n = 15) reciewed either 0.3 (0.3CAL), 0.5 ug CAUkgcd (O.SCAL)or vehicle (CON) s.e. for up to 12 days. During treatment the animals ware housed In metabolic cages for daily determination of calcium and hydroxyprolina excretion. Serum calcium and ostaocalcin were determined every other day. Graups of 5 rats from each treatment ware sacrificed on day 4, 8 and 12(tO). Calcium snd anorganic phosphate content of the left femur was analysad by ashing. Half of the right femur was embedded for histological analysis and the other half was used for the hydroxyproline assay. During treatment (O.BCAL, 0.5CAL) serum calcium rosa up to 3.4 mmoles/l (day 4), osteocalcin rose from 38 ng/ml (CON) to 84 ng/ml (MCAL) and 174 n&ml. Daily calcium excretion In treated groups was 2 - 3x highar than In controls, bydro~pr~llna excretion remained unchenged. Histology demonstrated Increased ostaoclast numbers at day 4. Aftar 10 teold seams were found slang with an creased. Femur calcium slSnific~nly lowered (O.SCAL)st day 8,lO and 12. Although the aboolutr amoun? of femur hydroxyroline decreased, there was on incraesa In relative hydroxyprollne content of the bone. Our data suggest, that high dose CAL treatment raises osteoclast numbers and mobilizes bona calcium Prior to stimulating prollferatlon and activity of osteoblasts.
140 PARATHYROID AUTOANTIBODIES AND MHC CLASS II ANTIGEN EXPRESSION IN HYPERPARATHYjkOl~I;M * hlin*. L. Klar GjC.luCi e? J. ,Rastad*, Department of Surgery* and Clinical Im\::nndigy, Uppsala University Hospital, Sweden. Autoimmune diseases are often associated with induction of MHC class II antigens and production of autoantibodies against cellmembrane stn~ctuns in the affected tissue. Frozen pnrathyroid biopsies from 77 patients with primary hyperparathyroidism (HPT), 17 patients with uremic HFT and I2 euparathyroid individuals were sectioned and inmrmohistochemically stained with monoclonal anti-WA-DR, -DQ and -DP antibodies using the PAP-technique, and with anti-Let1 4 and antiLeu 12 for detection of T- and B-lymphocytes. Sera from 27 of the patients with adenoma, 11 with primary hyp&plasia, 5 with urdmic HPT and 18 healthy contrck werr used for PAP-staining of frozen sections from human pxathyroicl tissue transplanted tonude mice for 4 weeks. The parenchyma of 13/54 adenomas and 8123, primary hyperplastic glands but none of the normal pamthyroids or the uremic glands showed immunoreactivity wit! the I-ILA-DR antibody. Anti-HLA-DQ and -DP yielded sunilar staining patterns. 22/27 sera from patients with adenomas gave ;I clsa~ immunoreactivity and 13 of these came from patients with I-ILA. DR expressing parathyroid glands. S/1 1 serd from patientr wirh p.-imary hyperplasia were positive and 4 of these came from -DK positive patients. 2/5 sera from uremic patients were postive while all control sera were negative. The results imply that autoimmune phenomena could he involved in the pathogenesis of primaryHPT.
109
CALCIfONIN IN THE TREATWENT OF POSTNENOPAUSAL OSTEOPOROSIS. C.Cepollaro, S.Gonnelli, M.Montomoli, A.Camporeale, D.Acrnusdei, F.Zacchei, M.S.Campagna, C.Gennari. Instituteof Internal Medicine,Universityof Siena, Italy. Some studies,performedwith extractivegrowth hormone (GH), suggested its use as stimulatorof bone formation in the treatment of postmenopausalosteoporosis (PMO). the efficacy of GH therapy in PM0 is still However, debated. Now a DNA recombinant human GH (rhGH) is available for clinical use. The present single blind study has been performed in order to compare the effects of rhGH given alone or in combination with salmon calcitonin (sCT) on bone turnover and bone mass in patients with PMO. 30 women (age 60.824 SD) with established PM0 were divided into 3 groups of 10 and randomly assigned to 3 treatment sequences:1) rhGH (12 Iv/d) S.C. for 7 days, followedby sCT (50 IU/d) S.C. for 21 days, and by 61-days free; 2) rhGH for 7 days, followed by placebo (Pl) for 21 days, and by Cl-days free; 3) Pl for 7 days, followedby sCT for 21 days, and by 61-days free. Each cycle was repeated4 times. At days 0, 8, 29 and 90 of each cycle, serum osteocalcin (BGP) and urinary excretion of Ca (CaU) and hydroryproline (IlOP were measured. QMD of thn lumbar sDine. femoral shaft, and distal radius was measurod,by IX%, defore and after 6 and 12 months. No significantchanges in EMD were observed wlthin and among thn 3 groups. With regard to the indaxos of bane turnover,a constant and significant increase2n BGP, MOP und Call wao detuctod during tha rhGl1 periods. HOP, as expected, significanllydecrescd during sC!Tperiods. Our study suggests that rhGtl acts as an activator of bone turnover rather than as o stimulatorof bone formation.
139 ACUTE EFFECT OF SHELL FORMATION ON 1,25 (OH),D, AND OSTEOCALCIN BLOOD LEVELS AND ON INTESTINAL AND UTERINE CALSINDIN mRNA IN HENS. X&&s. K. Allewatzt. I&W*, v-; INRA, Nouzilly, F37380; ‘LEGENDO, Leuven, S3oo0, ’ Cambridge, UK. Tha time-course of the changes in ionized calcium, 1,25(0H),D,, and osteocalcln blood levels and in the intestinal and uterine culblndin mRNA were studied In hens following suppression and resumption ol shell formallon and throughout the leylng cycle in hens laying hard-shelled or shell-lees eggs, In hens laying hard-shelled eggs circulallng l,25(OHJr& and osleocalcin, and uterinecalbindln mRNA were higher during the period of shell secretion but no differences were observed for intestinal calblndin mRNA. Plasma 1,25(OH)&, and osteocalcin were decreased 3h and 6h after suppression of shell formation. Uterine calbindin mRNA was depressed to 54 % of its initial value lh after egg expulsion and remained al a WY low level when eoa exoulslon was continued for 8 days. lntesllnal calblndin mRNA level was n6l affectad by suppression of shell formation. Calbindln levels decreased In both tissues only when premature ewpulslons of the egg were carried out for several da&When egg shell formallon resumed in hens previously laying sheRI-less eggs, the concenlratlon of 1,25(OH),4, osteocalcin and of inteslinal calbindin mANA increased toward Ihe end of shell formation but uterine calblndln and its mRNA were enhanced , earlier, al the onset on shell lormation. These Increases were maintalned at the uterine level in contrast lo the intestine &en hens were parathyroideclomised before shell resumption and when, consequently 1,25(0H),DS decreased. Feeding hens a tow calcium diet caused a large Increase in plasma 1,25(OH),DJ and osleocalcin. Ionized calcium levels tended to show reciprocal changes lo 1,25(0H),D,. However in hypercalcemic hens laying shelt-less eggs and fed a 3.5% calcium diet, the plasma 1,25(OH),D, was al a high level after ovulationand diminishedthereafter. In conctuslon, this last observation suggest that In hens, 1,25(OH),D, production Is regulated, in addilion lo Ihe stimulalion Invoking decreased C&I” and increased PTH, by anolher possibly endocrine factor linked lo ovulation. Acute changes in plasma 1,25(OH),D, and osleocalcin were parallel suggesting a relationship between these oaramelers. In contrast intestinalmRNA was stable posdbly as a consequ&ce of the transepilhelial calcium transport. Large daily increase in utetine calbindinmRNA occured irrespectively Of 1,25(OHJ,D, ievels and coincided with uterine calcium transpotl indicating that its regulation involves a calcium flux dependent factor.
138 FFECTS OF tllGW DOSE CALCITRIOL
ON RAT BONE
Scharing Resaarch, Allgemaine Pharmakologie. Berlin. FRG. We s&died time dependent changes in bone histology and re5paCtiVe biochemical markers of bone turnover during high dOSacalcltriol treatment (CAL) in adult (b.w.: 200 - 2409) female rats. 3 groups of weight matched animals (n = 15) reciewed either 0.3 (0.3CAL), 0.5 ug CAUkgcd (O.SCAL)or vehicle (CON) s.e. for up to 12 days. During treatment the animals ware housed In metabolic cages for daily determination of calcium and hydroxyprolina excretion. Serum calcium and ostaocalcin were determined every other day. Graups of 5 rats from each treatment ware sacrificed on day 4, 8 and 12(tO). Calcium snd anorganic phosphate content of the left femur was analysad by ashing. Half of the right femur was embedded for histological analysis and the other half was used for the hydroxyproline assay. During treatment (O.BCAL, 0.5CAL) serum calcium rosa up to 3.4 mmoles/l (day 4), osteocalcin rose from 38 ng/ml (CON) to 84 ng/ml (MCAL) and 174 n&ml. Daily calcium excretion In treated groups was 2 - 3x highar than In controls, bydro~pr~llna excretion remained unchenged. Histology demonstrated Increased ostaoclast numbers at day 4. Aftar 10 teold seams were found slang with an creased. Femur calcium slSnific~nly lowered (O.SCAL)st day 8,lO and 12. Although the aboolutr amoun? of femur hydroxyroline decreased, there was on incraesa In relative hydroxyprollne content of the bone. Our data suggest, that high dose CAL treatment raises osteoclast numbers and mobilizes bona calcium Prior to stimulating prollferatlon and activity of osteoblasts.
140 PARATHYROID AUTOANTIBODIES AND MHC CLASS II ANTIGEN EXPRESSION IN HYPERPARATHYjkOl~I;M * hlin*. L. Klar GjC.luCi e? J. ,Rastad*, Department of Surgery* and Clinical Im\::nndigy, Uppsala University Hospital, Sweden. Autoimmune diseases are often associated with induction of MHC class II antigens and production of autoantibodies against cellmembrane stn~ctuns in the affected tissue. Frozen pnrathyroid biopsies from 77 patients with primary hyperparathyroidism (HPT), 17 patients with uremic HFT and I2 euparathyroid individuals were sectioned and inmrmohistochemically stained with monoclonal anti-WA-DR, -DQ and -DP antibodies using the PAP-technique, and with anti-Let1 4 and antiLeu 12 for detection of T- and B-lymphocytes. Sera from 27 of the patients with adenoma, 11 with primary hyp&plasia, 5 with urdmic HPT and 18 healthy contrck werr used for PAP-staining of frozen sections from human pxathyroicl tissue transplanted tonude mice for 4 weeks. The parenchyma of 13/54 adenomas and 8123, primary hyperplastic glands but none of the normal pamthyroids or the uremic glands showed immunoreactivity wit! the I-ILA-DR antibody. Anti-HLA-DQ and -DP yielded sunilar staining patterns. 22/27 sera from patients with adenomas gave ;I clsa~ immunoreactivity and 13 of these came from patients with I-ILA. DR expressing parathyroid glands. S/1 1 serd from patientr wirh p.-imary hyperplasia were positive and 4 of these came from -DK positive patients. 2/5 sera from uremic patients were postive while all control sera were negative. The results imply that autoimmune phenomena could he involved in the pathogenesis of primaryHPT.
109