RESUMO
An acute systemic acidosis in vitamin D depleted rats that was induced by ammonium chlroide feeding resulted in defective biological hydroxylation of 25-hydroxycholecalciferol to 1,25-dihydroxycholecalciferol. Impaired enzymatic hydroxylation occurred despite the presence of either hypophosphatemia or hypocalcemia. The data suggest that acidosis interfers with the adaptive enzymatic control of 25-hydroxycholecalciferol metabolism in the vitamin D depleted state.
Assuntos
Acidose/metabolismo , Hidroxicolecalciferóis/metabolismo , Acidose/sangue , Acidose/induzido quimicamente , Cloreto de Amônio , Animais , Cálcio/sangue , Di-Hidroxicolecalciferóis/biossíntese , Hidroxicolecalciferóis/sangue , Mucosa Intestinal/metabolismo , Masculino , Fósforo/sangue , RatosRESUMO
The role of sodium in intestinal calcium transport was investigated in everted rat intestine. Ethacrynic acid, but not ouabain, inhibited calcium transport. However, ouabain did inhibit net water transport and, therefore, sodium transport, establishing the dissociation of the two transport processes. In addition to a magnesium-dependent adenosine triphosphatase (activated by sodium and potassium), a phosphatase dependent on sodium and calcium was localized to the lateral and basal membrane fractions of the mucosal cell. Activity of the latter phosphatase, similar to calcium transport in intact tissue, was inhibited by ethacrynic acid and not by ouabain. Sodium, therefore, may participate in the calcium transport process by activating an enzyme complex, dependent on adenosine triphosphate, that mediates calcium transport.
Assuntos
Cálcio/metabolismo , Duodeno/metabolismo , Íleo/metabolismo , Sódio/metabolismo , Adenosina Trifosfatases/análise , Adenosina Trifosfatases/metabolismo , Animais , Transporte Biológico Ativo/efeitos dos fármacos , Cálcio/farmacologia , Membrana Celular/enzimologia , Duodeno/enzimologia , Ácido Etacrínico/farmacologia , Íleo/enzimologia , Magnésio/farmacologia , Ouabaína/farmacologia , Ratos , Sódio/farmacologia , Equilíbrio Hidroeletrolítico/efeitos dos fármacosRESUMO
When compared to that of normal animals, calcium-binding protein activity of duodenal mucosa obtained from uremic rats was decreased. There was no change in this activity after vitamin D(3), therapy. In contrast, prior treatment with 25-hydroxycholecalcijerol resulted in increased transport of calcium-45 and calcium-binding protein activity in the intestine.
Assuntos
Cálcio/metabolismo , Absorção Intestinal , Ligação Proteica , Uremia/metabolismo , Animais , Colecalciferol/farmacologia , Colecalciferol/uso terapêutico , Doença Crônica , Duodeno , Mucosa Intestinal/metabolismo , Masculino , Ratos , Estimulação Química , Uremia/tratamento farmacológicoRESUMO
In vitro evidence presently favors a direct osteolytic effect of biologically active vitamin D metabolites. Studies were designed to evaluate the effect of 25-hydroxycholecalciferol (25OHD3) on bone collagen and mineral maturation in vivo and its dependence on parathyroid hormone (PTH). After treatment of sham-operated control and parathyroidectomized (PTX) mature rats with either 25OHD3 or an oil vehicle for 2 wk, tibial bone mineral-collagen maturation was quantitated by bromoform-toluene density gradient fractionation techniques. Intestinal calcium absorption was measured by in vivo 45Ca transport procedures. In contrast to the control group, the response to 25OHD3 of PTX rats was dramatic. Bone mineral and matrix maturation were both accelerated by 25OHD3 treatment without concomitant reduction in total bone mineral or collagen content or changes in the intestinal calcium absorption. These observations support the premise that biologically active vitamin D metabolites stimulate bone tissue maturation, and that PTH is not required in this regard.
Assuntos
Desenvolvimento Ósseo/efeitos dos fármacos , Osso e Ossos/metabolismo , Hidroxicolecalciferóis/farmacologia , Glândulas Paratireoides/fisiologia , Hormônio Paratireóideo/fisiologia , Animais , Matriz Óssea/metabolismo , Reabsorção Óssea/metabolismo , Cálcio/metabolismo , Colágeno/biossíntese , Colágeno/metabolismo , Di-Hidroxicolecalciferóis/farmacologia , Duodeno/metabolismo , Feminino , Hidroxicolecalciferóis/sangue , Hidroxiprolina/metabolismo , Absorção Intestinal , Minerais/metabolismo , Glândulas Paratireoides/cirurgia , Fósforo/metabolismo , Ratos , Estimulação Química , Vitamina D/farmacologiaRESUMO
The effect of chronic renal disease on bone matrix and mineral maturation was evaluated in rats with experimental renal insufficiency of 2-11 wk duration utilizing bromoform-toluene gradient fractionation of bone powder, pulse labeling experiments with (45)Ca and proline-(3)H differential extraction, and X-ray diffraction techniques.Maturation defects in both collagen and mineral ((45)Ca) metabolism were documented as early as 2 wk after the induction of uremia, when total bone calcium, inorganic phosphate, and hydroxyproline content were unchanged. The maturational defect progressed with advancing uremia despite insignificant changes in plasma pH and calcium, and normal bone carbonate levels. Although circulating levels of 25-hydroxycholecalciferol were significantly lower than normal in the uremic animals, pretreatment with either this vitamin D metabolite or vitamin D(3) itself failed to alter the observed changes in skeletal maturation.
Assuntos
Matriz Óssea/metabolismo , Cálcio/metabolismo , Colágeno/metabolismo , Falência Renal Crônica/metabolismo , Animais , Desenvolvimento Ósseo , Matriz Óssea/efeitos dos fármacos , Osso e Ossos/metabolismo , Cálcio/sangue , Isótopos de Cálcio , Carbonatos/metabolismo , Centrifugação com Gradiente de Concentração , Modelos Animais de Doenças , Feminino , Concentração de Íons de Hidrogênio , Hidroxicolecalciferóis/sangue , Hidroxiprolina/metabolismo , Fosfatos/metabolismo , Ratos , Ratos Endogâmicos , Fatores de Tempo , Trítio , Vitamina D/farmacologia , Difração de Raios XRESUMO
Proline-(14)C was administered to five adult rhesus monkeys, and the degradation of collagen was followed by the excretion of hydroxyproline-(14)C. The results suggested the presence of at least three separate pools of collagen with half-lives of 1 to 2, 2 to 3, and 50 to 70 days. The monkeys were killed after 44 days; at that time the specific activity of the hydroxyproline-(14)C in urine was found to be four to five times that of the hydroxyproline in soluble collagen and 81 to 93% that of hydroxyproline in insoluble collagen. The relationships between urinary hydroxyproline and the degradation of collagen were similar to those previously demonstrated in rats. Parathyroid hormone was administered daily to two of the monkeys from the 27th to the 44th day of the study. The parathyroid hormone increased the amount of hydroxyproline-(14)C excreted, but there was no significant change in the specific activity of the urinary hydroxyproline-(14)C. Since under the conditions of the experiment insoluble collagen was the only possible source of hydroxyproline-(14)C of relatively high specific activity, the results indicated that parathyroid hormone directly or indirectly increased the degradation of insoluble collagen. The results also suggested that parathyroid hormone increased the degradation of soluble collagen, but the relative magnitude of this effect was not clearly established.
Assuntos
Colágeno/metabolismo , Hidroxiprolina/urina , Hormônio Paratireóideo/farmacologia , Prolina/farmacologia , Animais , Cálcio/sangue , Isótopos de Carbono , Haplorrinos , RatosRESUMO
Bone mineral and matrix maturation in chronically uremic, nonacidotic rats were investigated after 25-hydroxcholecalciferol (25OHD) and/or dichloromethylene diphosphonate (C12MDP) therapy utilizing bromoform-toluene density gradient fractionation and X-ray diffraction analyses. The bromoform-toluene density gradient analyses demonstrated that the progressive accumulation of less dense, more immature bone characteristic of progressive uremia was reversed by 25OHD and/or C12MDP therapy for a 2-wk period, and that after 4 wk of therapy the maturational profile of bones from chronically uremic animals treated with 250HD and/or C12MDP was comparable to that from nonuremic littermates. X-ray diffraction analysis revealed that by the 4th wk of therapy with 25OHD and C12MDP both the degree of crystallinity and the crystal size/perfection parameters in the uremic bones were comparable to those of nonuremic, pair-fed control littermates. Treatment for 4 wk with 25OHD resulted in enlarged and/or more perfect apatite crystallites, while C12MDP alone slightly inhibited crystal growth and/or perfection after 2 wk of treatment. Soft tissue calcification was diminished in uremic animals treated for 4 wk with C12MDP or a combined C2MDP/25OHD regimen, the latter being much more effective in this regard. The accumulated data in this study support the premise that the attendant accelerated bone resorption, soft tissue calcification, and abnormal mineralization and maturation of the skeletal tissue, well documented to characterize experimental ranal insufficiency, may be alleviated with therapeutic dosages of 25OHD and/or C12MDP.
Assuntos
Distúrbio Mineral e Ósseo na Doença Renal Crônica/tratamento farmacológico , Hidroxicolecalciferóis/uso terapêutico , Organofosfonatos/uso terapêutico , Compostos Organofosforados/uso terapêutico , Animais , Osso e Ossos/metabolismo , Cálcio/metabolismo , Creatinina/sangue , Quimioterapia Combinada , Feminino , Hidroxiprolina/urina , Rim/metabolismo , Músculos/metabolismo , Fosfatos/metabolismo , Ratos , Uremia/metabolismoRESUMO
Hypocalcemia and resistance to exogenous parathyroid hormone have been reported in several clinical states associated with magnesium deficiency. On the basis of such observations, it has been suggested that magnesium depletion per se may result in impaired responsiveness of the adenyl cyclase-adenosine 3',5'-monophosphate (3',5'-AMP) system. To test this hypothesis, 4 wk old male parathyroidectomized rats were maintained on normal or magnesium-deficient diets for 4 wk and their responses to parathyroid hormone compared. Serum magnesium and calcium fell progressively in the magnesium-deficient group. Despite clinical and biochemical evidence of severe magnesium deficiency in these animals, renal production and excretion of 3',5'-AMP in response to parathyroid hormone was normal both in vitro and in vivo. Additionally, administration of either dibutyryl 3',5'-AMP or parathyroid extract to fasting magnesium-depleted rats produced a normal increase in serum calcium. Parathyroid hormone infusion studies demonstrated normal renal and skeletal responsiveness as measured by urinary excretion of calcium, magnesium, phosphate, and hydroxyproline. These data show that the effect of parathyroid hormone on 3',5'-AMP formation and excretion, the responsiveness of skeletal tissue to 3',5'-AMP, and the renal and skeletal system responses to parathyroid hormone are not altered by pure magnesium deficiency in the parathyroidectomized rat.
Assuntos
Deficiência de Magnésio/fisiopatologia , Hormônio Paratireóideo/farmacologia , Monofosfato de Adenosina/urina , Animais , Osso e Ossos/efeitos dos fármacos , Cálcio/sangue , Cálcio/urina , AMP Cíclico/biossíntese , GMP Cíclico/biossíntese , Fêmur , Nucleotídeos de Guanina/urina , Hidroxiprolina/urina , Rim/metabolismo , Magnésio/sangue , Magnésio/urina , Masculino , Glândulas Paratireoides/cirurgia , Fosfatos/sangue , Fosfatos/urina , RatosRESUMO
X-ray diffraction analysis of bone from chronically uremic but nonacidotic rats with normocalcemia and hyperphosphatemia revealed smaller apatite crystals and an increase in the X-ray amorphous mineral fraction when compared to age-matched, pair-fed control animals, indicating less advanced mineral maturation in the uremic animals. Studies in animals with varied degrees of chronic renal insufficiency revealed a progression of the bone crystal maturational defect with advancing uremia.
Assuntos
Desenvolvimento Ósseo , Osso e Ossos/metabolismo , Minerais/metabolismo , Uremia/metabolismo , Animais , Apatitas , Colágeno/metabolismo , Feminino , Hidrazinas , Métodos , Minerais/análise , Nefrectomia , Ratos , Tíbia/crescimento & desenvolvimento , Tíbia/metabolismo , Difração de Raios XRESUMO
To analyze their relative effects on premenopausal bone mass, we have studied the impact of lifelong estrogen exposure, assessed by an estrogen score (ES; computed on age at menarche, average length of menstrual cycles since menarche, and use of birth control pills), heredity, and some environmental factors on vertebral bone density (VBD), of 63 premenopausal women (age, 19-40 yr). Compared with women with normal bone density (Z score > -1), subjects with low VBD (Z score < -1) had significantly lower ES (15.1 +/- 3.9 vs. 18.7 +/- 2.4, P = 0.001), higher age at menarche (13.8 +/- 1.7 vs. 12.6 +/- 1.4 yr, P = 0.005), and lower serum estradiol (46.9 +/- 37 vs. 86.6 +/- 57 pg/ml, P = 0.023) and estrone levels (107.4 +/- 60 vs. 178.8 +/- 9.0 pg/ml, P = 0.05). Likewise, women in the lowest quartile for VBD had significantly lower ES (15.3 +/- 4.5 vs. 18.1 +/- 2.7, P = 0.006) and higher age at menarche (13.9 +/- 1.9 vs. 12.8 +/- .4, P = 0.02) than those in the upper three quartiles. A higher proportion of subjects with irregular menses (52 vs. 23%, P = 0.03) and a positive family history of osteoporosis (86 vs. 61%, P = 0.04) was found in the low VBD group compared with subjects with normal VBD. VBD correlated positively with ES (r = 0.44, P = < 0.001) and negatively with age at menarche (r = -0.30, P = 0.03) by simple linear regression, whereas no correlation was found between VBD and age, body mass index, parity, lactation, physical activity, sunlight exposure, and dietary calcium and vitamin D intakes. The correlation between VBD and ES improved after correcting for the effect of all the other variables by partial correlation analysis (Pearson partial r = 0.57, P = < 0.01), which also disclosed a significant contribution of dietary calcium to VBD. However, ES was the only significant independent determinant of VBD, by stepwise multiple regression analysis (R2 = 0.24). Therefore, premenopausal estrogen exposure, and possibly genetic predisposition, rather than environmental factors, are the major determinants for the development of peak bone mass before menopause.
Assuntos
Estrogênios/fisiologia , Menopausa , Osteoporose/genética , Adulto , Fatores Etários , Densidade Óssea , Dieta , Feminino , Humanos , Fatores de Risco , População BrancaRESUMO
The metabolic fate of intravenously injected vitamin D(3)-1,2-(3)H (D(3)-(3)H) was studied in two normal individuals on chronic phenobarbital therapy. Silicic acid column chromatography of lipid-soluble plasma extracts obtained serially for 96 hr after D(3)-(3)H injection demonstrated a decreased plasma D(3)-(3)H half-life and increased conversion to more polar metabolites. The polar metabolites formed included several with chromatographic mobility similar to known biologically inactive vitamin D metabolites and one with chromatographic mobility identical to 25-hydroxycholecalciferol. Disappearance of this latter material was also accelerated. A child with rickets and a normal volunteer studied before and after a 2 wk course of phenobarbital therapy demonstrated similar alterations in D(3)-(3)H metabolism. When liver microsomes from 3-wk-old Sprague-Dawley rats treated with phenobarbital were incubated with D(3)-(3)H, polar metabolites were produced with chromatographic mobility similar to the plasma D(3)-(3)H metabolites from phenobarbital-treated humans. Similar incubations employing 25-hydroxy-cholecalciferol-26-27-(3)H as the substrate also demonstrated an increased conversion to polar metabolites. The data suggest that the reported increased incidence of osteomalacia observed in patients on chronic anticonvulsant therapy may be the result of an accelerated conversion of vitamin D and its active metabolite, 25-hydroxycholecalciferol, to polar metabolites by druginduced liver microsomal enzymes.
Assuntos
Colecalciferol/metabolismo , Fenobarbital/farmacologia , Adulto , Animais , Anticonvulsivantes/efeitos adversos , Colecalciferol/sangue , Cromatografia em Camada Fina , Indução Enzimática , Feminino , Meia-Vida , Humanos , Hidroxicolecalciferóis/metabolismo , Taxa de Depuração Metabólica , Microssomos Hepáticos/enzimologia , Microssomos Hepáticos/metabolismo , Pessoa de Meia-Idade , Oxigenases de Função Mista/metabolismo , Osteomalacia/induzido quimicamente , Fenobarbital/uso terapêutico , Fenitoína/farmacologia , Ratos , TrítioRESUMO
The fate of an intravenous dose of tritiated vitamin D(3) was studied in seven normal subjects, four children with vitamin D-resistant rickets, and four adults with a familial history of vitamin D-resistant rickets and persistent hypophosphatemia. An abnormal metabolism of vitamin D in vitamin D-resistant rickets was defined and characterized by a decrease in the plasma fractional turnover rate, a marked increase in plasma water-soluble metabolites, and a relative decrease in the conversion of vitamin D to a polar, biologically active metabolite. Alterations in vitamin D metabolism in the adults with persistent hypophosphatemia were similar but less severe than those of affected children with vitamin D-resistant rickets. It is tentatively concluded that the abnormalities in vitamin D metabolism documented in patients with vitamin D-resistant rickets and familial hypophosphatemia may account for the observed osseous and biochemical changes.
Assuntos
Colecalciferol/metabolismo , Hipofosfatemia Familiar/metabolismo , Raquitismo/metabolismo , Adolescente , Adulto , Fosfatase Alcalina/sangue , Cálcio/sangue , Criança , Colecalciferol/análise , Colecalciferol/sangue , Colecalciferol/urina , Cromatografia , Cromatografia em Gel , Cromatografia em Camada Fina , Fezes/análise , Feminino , Humanos , Hipofosfatemia Familiar/sangue , Hipofosfatemia Familiar/urina , Masculino , Fósforo/sangue , Raquitismo/sangue , Raquitismo/urina , TrítioRESUMO
The absorption and metabolism of vitamin D(3)-(3)H was studied in eight patients with chronic renal failure. Although the intestinal absorption of vitamin D(3)-(3)H was normal, the metabolic fate of the vitamin was abnormal as characterized by a twofold increase in fractional turnover rate, an abnormal accumulation of biologically inactive lipid-soluble metabolites, and the urinary excretion of both vitamin D(3)-(3)H and biologically inactive metabolites. Neither alterations in water-soluble vitamin D(3) metabolites nor qualitative abnormalities in protein-binding of vitamin D(3) were observed in the uremic subjects. Although hemodialysis proved ineffectual in reversing the observed abnormalities in vitamin D(3) metabolism and excretion, renal homotransplantation was completely successful in this regard. These experiments support the conclusion that the resistance to therapeutic doses of vitamin D often seen in patients with chronic renal failure and renal osteodystrophy results from an acquired defect in the metabolism and excretion of vitamin D.
Assuntos
Colecalciferol/metabolismo , Falência Renal Crônica/metabolismo , Adulto , Fosfatase Alcalina/sangue , Animais , Bioensaio , Nitrogênio da Ureia Sanguínea , Clorofórmio , Colecalciferol/análise , Colecalciferol/sangue , Colecalciferol/urina , Cromatografia , Eletroforese , Fezes/análise , Feminino , Glomerulonefrite/metabolismo , Humanos , Falência Renal Crônica/sangue , Transplante de Rim , Masculino , Síndrome Nefrótica/metabolismo , Proteinúria , Pielonefrite/metabolismo , Ratos , Diálise Renal , Transplante Homólogo , TrítioRESUMO
UNLABELLED: To investigate the effectiveness of calcitonin treatment of postmenopausal osteoporosis in relation to bone turnover, we examined 53 postmenopausal osteoporotic women before and after one year of therapy with salmon calcitonin (sCT), at the dose of 50 IU every other day. Baseline evaluation revealed that 17 (32%) patients had high turnover (HTOP), and 36 (68%) normal turnover osteoporosis (NTOP) as assessed by measurement of whole body retention (WBR) of 99mTc-methylene diphosphonate. The two groups did not differ in terms of bone mineral content (BMC) measured by dual photon absorptiometry at both lumbar spine and femoral diaphysis. However, HTOP patients had higher levels of serum osteocalcin (OC) and urinary hydroxyproline excretion (HOP/Cr). Multivariate regression analysis showed no correlation between parameters of bone turnover (WBR, OC, HOP/Cr) and both femoral and vertebral bone density; the latter being negatively correlated only with the years elapsed since menopause (R2 = 0.406). Treatment with sCT resulted in a significant increase of vertebral BMC in the 53 patients taken as a whole group (+/- 7%, P less than 0.001). When the results obtained in HTOP and NTOP were analyzed separately, only those with HTOP showed a marked increment of spinal BMC (+22%, P less than 0.001), NTOP subjects neither gained nor lost bone mineral during the study. Femoral BMC decreased in the whole group after sCT therapy (-3%, P less than 0.003). However, HTOP patients maintained initial BMC values, whereas those with NTOP lost a significant amount of bone during the study period (-5%, P less than 0.001). The increase of vertebral bone mass was associated with a marked depression of bone turnover detectable in both subsets of patients and in the whole group. IN CONCLUSION: (a) assessment of bone turnover cannot help predict the severity of bone loss in postmenopausal osteoporosis; (b) calcitonin therapy appears to be particularly indicated for patients with high-turnover osteoporosis, resulting in a net gain of bone mineral in the axial skeleton and a slowing of bone loss in the appendicular bones.
Assuntos
Reabsorção Óssea/efeitos dos fármacos , Calcitonina/uso terapêutico , Menopausa/fisiologia , Osteoporose/fisiopatologia , Envelhecimento , Biomarcadores , Proteínas de Ligação ao Cálcio/análise , Feminino , Humanos , Hidroxiprolina/análise , Pessoa de Meia-Idade , Minerais/metabolismo , Osteocalcina , Osteoporose/tratamento farmacológico , Osteoporose/metabolismo , Prolina/análiseRESUMO
Human osteoblast cultures (hOB) were examined for the production of interstitial collagenase, tissue inhibitor of metalloproteinases (TIMP), and gelatinolytic enzymes. Cells were isolated by bacterial collagenase digestion of trabecular bone (vertebra, rib, tibia, and femur) from 11 subjects (neonatal to adult). Confluent cultures were exposed to phorbol 12-myristate 13-acetate, PTH, PGE2, epidermal growth factor, 1,25(OH)2 vitamin D3, recombinant human IL-1 beta, and dexamethasone. Collagenase and TIMP were assayed immunologically and also by measurements of functional activity. Collagenase was not secreted in significant quantities by human bone cells under any tested condition. Furthermore, collagenase mRNA could not be detected in hOB. However, hOB spontaneously secreted large amounts of TIMP for at least 72 h in culture. hOB TIMP was found to be identical to human fibroblast TIMP by double immunodiffusion, metabolic labeling and immunoprecipitation, Northern blot analysis, and stoichiometry of collagenase inhibition. SDS-substrate gel electrophoresis of hOB-conditioned media revealed a prominent band of gelatinolytic activity at 68 kD, and specific polyclonal antisera established its identity with the major gelatinolytic protease of human fibroblasts. Abundant secretion of gelatinolytic, but not collagenolytic, enzymes by hOB may indicate that human osteoblasts do not initiate and direct the cleavage of osteoid collagen on the bone surface, but may participate in the preparation of the bone surface for osteoclast attachment by removal of denatured collagen peptides. The constitutive secretion of TIMP may function to regulate metalloproteinase activity.
Assuntos
Inibidores Enzimáticos/metabolismo , Colagenase Microbiana/metabolismo , Osteoblastos/enzimologia , Pepsina A/metabolismo , Fosfatase Alcalina/análise , Reabsorção Óssea , Proteínas de Ligação ao Cálcio/metabolismo , Células Cultivadas , AMP Cíclico/biossíntese , Inibidores Enzimáticos/imunologia , Fibroblastos/metabolismo , Gelatinases , Humanos , Interleucina-1/farmacologia , Osteocalcina , Inibidores Teciduais de MetaloproteinasesRESUMO
Lithium chloride administration to growing rats, which resulted in circulating lithium levels of 1.4 meq/liter, was attended by significant suppression of bone mineralization and organic matrix synthesis as assessed by tetracycline labeling and histological quantitation of osteoid, respectively. These effects of lithium were not associated with changes in animal behavior, nor were there any significant differences in blood levels of calcium, phosphorus, alkaline phosphatase, creatinine, pH, or parathyroid hormone. The data suggest that lithium inhibition of bone mineralization is secondary to suppression of osteoid formation.
Assuntos
Osso e Ossos/efeitos dos fármacos , Lítio/farmacologia , Minerais/metabolismo , Animais , Desenvolvimento Ósseo , Osso e Ossos/metabolismo , Depressão Química , Feminino , RatosRESUMO
Anticonvulsant therapy of seizure disorders in man is associated with the development of complications involving bone and mineral metabolism including hypocalcemia, elevated serum immunoreactive parathyroid hormone levels, and increased amounts of unmineralized bone or osteoid. The latter has been attributed to a reduction in serum-25-hydroxycholecalciferol levels resulting from increased hepatic metabolism of vitamin D. Using an in vitro recycling hepatic perfusion system, we have demonstrated that 5 d of phenobarbital treatment increases the hepatic production of [(3)H]25-hydroxyvitamin D(3) (4.3+/-0.3 vs. 3.3+/-0.2%/h, P <0.025) without affecting the biliary excretion of radioactivity. Furthermore, rachitic livers perfused with blood obtained from animals treated with phenobarbital for 5 d also manifested an increase in [(3)H]25-hydroxyvitamin D(3) production (4.6+/-0.5 vs. 3.3+/-0.2%/h, P < 0.02). Addition of phenobarbital or its major metabolite, p-hydroxyphenobarbital, directly to the perfusion apparatus had no effect on [(3)H]25-hydroxyvitamin D(3) production. Phenobarbital treatment was also attended by a decrease in the intrahepatic content of [(3)H]vitamin D(3) (11.7+/-0.4 vs. 17.5+/-0.7 dpm/mg liver protein, P < 0.001) without alterations in the content of [(3)H]25-hydroxyvitamin D(3). The data collectively suggest that the increased hepatic conversion of [(3)H]vitamin D(3) to [(3)H]25-hydroxyvitamin D(3) attending phenobarbital treatment is secondary to stimulation of the hepatic 25-hydroxylation system(s) by a metabolite of phenobarbital other than p-hydroxyphenobarbital and/or by metabolic alterations resulting from phenobarbital therapy.
Assuntos
Colecalciferol/metabolismo , Fígado/metabolismo , Fenobarbital/farmacologia , Raquitismo/metabolismo , Animais , Relação Dose-Resposta a Droga , Técnicas In Vitro , Masculino , Microssomos Hepáticos/metabolismo , Mitocôndrias Hepáticas/metabolismo , Perfusão , RatosRESUMO
This study investigates the proliferative and osteogenic role of marrow stromal/osteoprogenitor cells in the development of the cortical bone deficit in ovariectomized (OVX) female rats. In vitro, clonal growth of marrow stromal cells from OVX rats was significantly impaired (vs. sham-operated controls). Yet in vivo, cells from sham-operated and OVX rats had equal osteogenic potential in several in vivo experimental situations, such as in intraperitoneally implanted millipore diffusion chambers and in intramuscular implants of marrow plus osteoinductive bone matrix (composite grafts). Long-term (6 mo) dihydrotachysterol (DHT) treatment of OVX rats enhanced their in vitro proliferative potential and clonal growth, as well as their osteogenic expression in composite grafts. The observation that the in vivo osteogenic performance of OVX rat marrow stromal cells was normal at extraosseous sites suggests that the mechanisms leading to osteopenia may involve an abnormality in cell-matrix interactions.
Assuntos
Doenças Ósseas/patologia , Medula Óssea/patologia , Di-Hidrotaquisterol/uso terapêutico , Ovariectomia , Animais , Doenças Ósseas/tratamento farmacológico , Doenças Ósseas/etiologia , Transplante de Medula Óssea , Matriz Óssea/fisiopatologia , Matriz Óssea/transplante , Divisão Celular , Células Cultivadas , Modelos Animais de Doenças , Feminino , Fibroblastos/patologia , Fibroblastos/fisiologia , Minerais/metabolismo , Osteogênese , Ratos , Ratos Endogâmicos , Células-Tronco/patologia , Células-Tronco/fisiologiaRESUMO
Vitamin D(3)-(3)H has been administered intravenously to seven normal subjects, three patients with biliary fistulas, and four patients with cirrhosis. Plasma D(3)-(3)H half-times normally ranged from 20 to 30 hours. in vivo evidence that a metabolic transformation of vitamin D occurs was obtained, and a polar biologically active vitamin D metabolite was isolated from plasma. Urinary radioactivity averaged 2.4% of the administered dose for the 48-hour period after infusion, and all the excreted radioactivity represented chemically altered metabolites of vitamin D. The metabolites in urine were mainly water-soluble, with 26% in conjugated form. From 3 to 6% of the injected radioactivity was excreted in the bile of subjects with T-tube drainage and 5% in the feces of patients having no T-tube. The pattern of fecal and biliary radioactivity suggested that the passage of vitamin D and its metabolites from bile into the intestine represents an essential stage for the fecal excretion of vitamin D metabolites in man. Abnormally slow plasma disappearance of vitamin D(3)-(3)H in patients with cirrhosis was associated with a significant decrease in the quantity and rate of glucuronide metabolite excretion in the urine.
Assuntos
Colecalciferol/metabolismo , Adulto , Animais , Bile/análise , Fístula Biliar/metabolismo , Bioensaio , Colecalciferol/sangue , Colecalciferol/urina , Cromatografia , Fezes/análise , Feminino , Humanos , Injeções Intravenosas , Cirrose Hepática/metabolismo , Masculino , Aves Domésticas , Ratos , Trítio , Vitamina D/metabolismoRESUMO
To test the hypothesis that mononuclear cells are stimulated to release interleukin 1 (IL-1) by bone fragments released in the bone microenvironment during the remodeling cycle, we have investigated the effects of bone matrix and some of its constituents on IL-1 secretin from peripheral blood mononuclear cells (PBMC). Increases in IL-1 activity were observed when either PBMC or adherent monocytes, but not lymphocytes depleted of monocytes, were co-cultured with either human or rat bone particles but not with latex particles of similar size. Co-culture of PBMC with bone particles in a transwell system where the cells were physically separated from the bone particles, or with osteoblast- or osteoclast-covered bone particles, did not stimulate IL-1 release, indicating that a physical contact between PBMC and the bone surface is required for eliciting IL-1 release. This was confirmed by the finding of a lower stimulatory effect of bone particles pretreated with etidronate, a bisphosphonate which decreases the bone binding capacity of PBMC. Constituents of bone matrix, such as collagen fragments, hydroxyproline, and, to a lesser extent, transforming growth factor-beta, but not osteocalcin, alpha 2HS glycoprotein, fragments of either bone sialoprotein or osteopontin, and fibronectin, stimulated PBMC IL-1 release in a dose-dependent fashion. Collagen-stimulated IL-1 release was partially and specifically inhibited by a monoclonal antibody directed against the alpha 2 beta 1-integrin cell surface collagen receptor. These data demonstrate that products of bone resorption, known to be chemotactic for mononuclear cells, stimulate PBMC IL-1 activity. These findings may help explain previous documentation of increased IL-1 secretion by circulating monocytes obtained from patients with high turnover osteoporosis.