Role of 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) on intestinal inorganic phosphate (Pi) absorption in rats with normal vitamin D supply.
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Animals↗
Biomedical subjects
H Fleisch
Publications and source records attributed to H Fleisch.
The role of inhibitors and other factors in the pathogenesis of recurrent calcium-containing renal stones.
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Adult↗
Calcium absorption in diphosphonate-treated rats: effect of parathyroid function, dietary calcium and phosphorus.
1. The role of parathyroid hormone (PTH) and 1, 25-dihydroxy-cholecalciferol (1,25-(OH)2D3) in modulation of intestinal Ca absorption was studied in rats, using disodium ethane-1-hydroxy-1,1-diphosphonate (EHDP), which is known to reduce 1,25-(OH)2D3 formation. 2. EHDP decreased intestinal Ca absorption. This effect could be abolished by small amounts of 1,25-(OH)2D3, whereas even large doses of PTH were ineffective. EHDP also decreased Ca absorption in thyroparathyroidectomized (TPTX) rats. Therefore the effect of EHDP on 1,25-(OH)2D3 production is unlikely to be mediated through PTH. 3. The correction by PTH of the decreased Ca absorption in TPTX rats was inhibited by EHDP. Since EHDP inhibits formation of 1,25-(OH)2D3 the effect of PTH on Ca absorption is likely to be mediated through this vitamin D3 metabolite. 4. In normal rats both a low Ca and a low P diet stimulated Ca absorption. In EHDP-treated intact rats low Ca still stimulated Ca absorption, whereas the effect of low P abolished. This indicates that low Ca and low P diets affect Ca absorption through different mechanisms. 5. Intestinal adaptation to a low Ca diet was still observed in EHDP-treated TPTX rats. Thus, in the rat, intestinal adaptation to low Ca diet can occur without PTH.
Animals↗
Relation between bone mineralization, Ca absorption, and plasma Ca in phosphonate-treated rats.
Disodium ethane-1-hydroxy-1,1-diphosphonate (EHDP) is known to inhibit the crystallization of calcium phosphate salts in vitro. Large doses of EHDP administered in vivo inhibit skeletal mineralization, decrease intestinal calcium absorption, and produce hypercalcemia. In the present study, EHDP or one of 13 other phosphonates were given to rats at 10 mg P/kg-day sc for 7 days in order to better define the nature of the relationship between bone mineralization, intestinal absorption, and plasma calcium in the regulation of calcium homeostasis. Each of the phosphonates which inhibited skeletal mineralization in vivo also inhibited crystallization in vitro, but the converse was not true. A very close correlation was found between inhibition of skeletal mineralization, decreased intestinal calcium absorption, and slight hypercalcemia. A dose-response study with two compounds also revealed the same close correlation. It is argued that the impairment of intestinal calcium absorption in phosphonate-treated rats may represent a secondary homeostatic response to the primary effect of the drugs on bone mineralization. This response may be mediated by an elevation of a fraction of plasma calcium.
Animals↗
Role of parathyroid hormone in regulation of main calcium fluxes in rats.
The effect of calcium deprivation on the various calcium fluxes was studied in growing rats either sham-operated (SHAM), thyroparathyroidectomized (TPTX), or thyroparathyroidectomized and supplemented with parathyroid hormone (PTH) (TPTX + PTH). In SHAM rats a decrease in the net absorption of calcium (Vna) has no influence on calcemia or on bone formation (Vo+), but leads to an increase in bone resorption (Vo-). In TPTX rats a decrease in Vna induces a decrease in calcemia and in Vo+ but still causes an increase in Vo-. The same is true in TPTX + PTH rats although all the variables measured are increased. In TPTX rats, both without and with PTH, a linear correlation exists between calcemia and Vo+ suggesting that calcemia influences bone formation. Furthermore, it appears that PTH is important in regulating bone turnover, but that the adaptation of Vo- to a change in Vna can occur in the absence or in the presence of a constant amount of this hormone. The mechanism of regulating this adaptation of bone resorption is still unknown.
Animals↗
Effect of thyroparathyroidectomy of calcium metabolism in rats: role of 1,25-dihydroxyvitamin D3.
Thyroparathyroidectomy (TPTX) decreases plasma calcium, bone formation and resorption, and tubular reabsorption of calcium. It also reduces the production of 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] which very likely results in the decrease in the intestinal calcium absorption (Vna) observed after TPTX. We have examined whether the influence of TPTX on plasma calcium and bone calcium fluxes could be corrected by doses of 1,25-(OH)2D3 (2 X 13 pmol/day ip), which just normalize Vna. The study was made by calcium balance and 45Ca kinetics in rats receiving a normal supply of vitamin D3. The results show that in TPTX rats physiological doses of 1,25-(OH)2D3 increased plasma calcium and decreased plasma phosphate. Calcium retention was not fully corrected because 1,25-(OH)2D3 increased urinary calcium excretion. The deposition into and the release of calcium from bone were enhanced by 1,25-(OH)2D3 but remained lower than in pair-fed sham-operated animals. Thus, in as much as the diminished renal production of 1,25-(OH)2D3 in TPTX is entirely responsible for the reduced Vna, the decrease in 1,25-(OH)2D3 could only partly explain the effect of thyroparathyroidectomy on bone calcium fluxes.
Animals↗
Tubular localization of adaptation to dietary phosphate in rats.
The renal tubule of rats adapts its transport capacity for inorganic phosphate (Pi) in response to changes in dietary Pi intake. Tubular localization of this adaptation was studied in rats pair-fed for 10 days a low (LPD), i.e., 0.2 g/100 g, or a high (HPD), i.e., 1.8 g/100 g, phosphorus diet. Free-flow micropunctures under acute Pi infusion were made at elevated but similar filtered loads of Pi: LPD, 6.07 +/- 0.29 (n, 32); HPD 5.34 +/- 0.24 (n, 33) micromol/kidney per min. Fractional excretion of Pi (FEPi) for the whole kidney was: LPD, 0.26 +/- 0.02 (n, 32); HPD, 0.67 +/- 0.02 (n, 33) (P less than 0.001). The fraction of filtered Pi found at various puncture sites suggests that part of the adaptation to dietary Pi occurs in the early proximal tubule. An effect in the later portion of the proximal convoluted tubule cannot be excluded in the present experiments. Furthermore, dietary Pi affects markedly the difference in Pi delivery between distal tubule and final urine. This could be due to a greater adaptive response of deep nephrons. Alternatively, the results could also indicate that an adaptation with a secretory component takes place along the terminal part of the nephrons.
Adaptation, Physiological↗
Role of 1,25-dihydroxyvitamin D3 on intestinal phosphate absorption in rats with a normal vitamin D supply.
In vitamin D-deficient rats, impaired intestinal phosphorus (P) absorption can be corrected by 1,25-dihydroxyvitamin D(3)[1,25-(OH)(2)D(3)]. In the present study, it was investigated whether changes in 1,25-(OH)(2)D(3) production can influence intestinal P transport also in animals with a normal supply of vitamin D. The intestinal P absorption was evaluated in rats using both the in situ duodenal loop technique and the determination of the overall gastrointestinal absorption under three conditions known to influence the production of 1,25-(OH)(2)D(3): (a) variation in dietary P, (b) thyroparathyroidectomy (TPTX) with or without administration of parathyroid hormone (PTH), and (c) treatment with disodium ethane-1-hydroxy-1,1-diphosphonate (EHDP). In all circumstances changes in duodenal absorption paralleled the changes in the overall fractional absorption. (a) Lowering dietary P stimulated P absorption. (b) TPTX decreased P absorption. This effect was corrected either by the administration of PTH or by the administration of 1,25-(OH)(2)D(3). (c) EHDP, when given at a dose known to inhibit 1,25-(OH)(2)D(3) formation, decreased the duodenal P absorption in both intact and TPTX animals. This effect was corrected by 1,25-(OH)(2)D(3). In the TPTX-EHDP-treated animals, the administration of PTH did not rectify the low duodenal P absorption. These results support the thesis that, in rats with normal vitamin D supply, variations in the endogenous production of 1,25-(OH)(2)D(3) change the rate of P absorption. However, these changes are in such magnitude that they are of relatively small importance when compared to the effect of variation in the dietary intake of P. These results also strongly suggest that the action of PTH on duodenal P transport is mediated by its effect on 1,25-(OH)(2)D(3) production, inasmuch as the effect of the hormone is abolished after blocking the renal 1-hydroxylation with EHDP.
Animals↗
Effect of 1,25-dihydroxyvitamin D3 on the renal handling of Pi in thyroparathyroidectomized rats.
The kidney adapts its tubular capacity to transport inorganic phosphate (P(i)) according to the dietary supply of P(i) in both intact and thyropara-thyroidectomized (TPTX) rats. However, in TPTX rats the capability of the renal tubule to adapt to a high P(i) diet is diminished. In TPTX rats the production of the active vitamin D(3) metabolite, 1,25-dihydroxyvitamin D(3) [1,25-(OH)(2)D(3)], is also reduced. 1,25-(OH)(2)D(3) has been shown to have a marked effect on P(i) metabolism. Therefore the question arises whether the deficient production of 1,25-(OH)(2)D(3) contributes to the alteration of the tubular transport of P(i) observed in chronically TPTX rats. In the present investigation, vitamin D-replete rats were sham operated (SHAM) or thyroparathyroidectomized and then pair fed diets containing either 0.2 or 1.2 g/100 g P for 7 days. During this period, groups of SHAM and TPTX rats received i.p. 2 x 13 pmol/day of 1,25-(OH)(2)D(3), a dose which was shown to just normalize the decreased intestinal absorption of Ca and P(i) in TPTX rats. The capacity of tubular P(i) transport was then assessed by measuring the fractional excretion of P(i) (FEP(i)) at increasing plasma P(i) concentration ([P(i)](Pl)) obtained by acute infusion of P(i). The results show that in SHAM rats fed either P diet, 1,25-(OH)(2)D(3) has no effect on the renal handling of P(i). In TPTX rats fed 1.2 g/100 g P diet, 1,25-(OH)(2)D(3) increases FEP(i) over a wide range of [P(i)](Pl.) In TPTX rats fed a 0.2 g/100 g P diet, 1,25-(OH)(2)D(3) does not alter FEP(i) up to a [P(i)](Pl) of 3.0-3.5 mM, but does increase it at higher [P(i)](Pl.) In fact, on both diets TPTX rats supplemented with 1,25-(OH)(2)D(3) appear to have the same renal handling of P(i) as SHAM counterparts. The effect of 1,25-(OH)(2)D(3) was not associated with a change in urine pH or in urinary excretion of cyclic AMP and was maintained under marked extracellular volume expansion. It was associated with a rise in plasma calcium in the TPTX rats fed the high, but not the low, P diet. In TPTX rats fed 1.2 g/100 g P diet, 25-hydroxyvitamin D(3) in doses of 2 x 130 or 2 x 1,300 pmol/day i.p. did not increase FEP(i.)In conclusion, 1,25-(OH)(2)D(3) administered in physiological amounts to TPTX rats restores to normal the capability of the renal tubule to excrete P(i) and to adapt to large variation in dietary P(i). The results suggest that 1,25-(OH)(2)D(3) plays an important role in the regulation of the renal handling of P(i) and that the chronic change in the tubular capacity to transport P(i) after TPTX may be due to the decreased formation of 1,25-(OH)(2)D(3).
Animals↗
Aggregation of hydroxyapatite crystals.
A system to study the aggregation of hydroxyapatite crystals was developed. The effect of several factors (Ca2+ x Pi product, Ca2+/Pi ratio, pH, and various substances) were tested. Pb2+, Zn2+, Mg2+ and methyleneblue had only small effects; citrate inhibited aggregation. Pyrophosphate was a strong inhibitor and the diphosphonates disodium ethane-1-hydroxy-1,1-diphosphonate and disodium dichloromethylene diphosphonate were even more potent. The monophosphonate pentanemonophosphonate had no effect. Potent inhibition also occurred with glycosaminoglycans: heparin greater than hyaluronic acid greater than dermatan sulfate greater than chondroitin 4-sulfate greater than chondroitin 6-sulfate. Urine also showed high inhibitory activity. The inhibition of heparin but not that of hyaluronic acid, PPi or urine was abolished by egg white lysozyme. The effects described might be relevant in the normal mineralization process as well as in the mechanisms leading to pathological calcification, such as urinary stone formation.
Binding Sites↗
Disaggregation of hydroxyapatite crystals.
A system has been developed to measure quantitatively the disaggregation of hydroxyapatite crystals. Disaggregation was induced by pyrophosphate, ethane-1-hydroxy-1,1-diphosphonate, dichloromethylene diphosphonate, haparin and citrate. Hyaluronic acid stimulated aggregation at low concentrations and disaggregation at high concentrations. Lactate had no effect. The possible role disaggregation might play in the resorption of calcified tissues in vivo id discussed.
Binding Sites↗
Pyrophosphatase and ATPase of isolated cartilage matrix vesicles.
Some of the characteristics of the pyrophosphatase and ATPase activities studied in isolated cartilage matrix vesicles were found to be similar to those already reported for the solubilized and purified bone alkaline phosphatase. Thus, the pH optimum of the pyrophosphatase activity responded similarly to changes in the concentration of Mg2+, Ca2+, and PPi. Further, the ATPase activity was not activated by Ca2+ in the presence of an optimal Mg2+ concentration. It is proposed that a function of the alkaline phosphatase of matrix vesicles in vivo is to hydrolyze the substrates PPi, ADP, and ATP, which are known inhibitors of calcium phosphate precipitation.
Adenosine Triphosphatases↗
A new device for ultrafiltration. The ultrafiltrability of calcium and inorganic phosphate in human serum.
The various problems concerning the ultrafiltration of blood serum are outlined, and a novel ultrafiltration device, which overcomes all these problems is presented. Its main advantages are the following: (1) The deposition of protein on the filter is prevented by producing a flow across the filter surface. (2) The pCO2 remains constant, since the fluid is not in contact with a gas space. (3) The minimum serum volume is only 0.4 ml (4) Six samples can be processed simultaneously. (5) All commercially available filter types can be used. The machine, tested by measuring the ultrafiltrability of calcium, inorganic phosphate, sodium and chloride in human serum, proved to be a reliable laboratory tool. The precision of the determinations were comparable to respective serum analyses. The ultrafiltrabilities obtained, without correction for water content of serum, were 55.6 +/- 2.9% for calcium, 88.3 +/- 5.0% for inorganic phosphate, 95 +/- 1.7% for sodium, and 103 +/- 0.7% for chloride (means +/- S.D.).
Adult↗
Renal tubular adaptation to dietary phosphorus.
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Adaptation, Physiological↗
The effect of several diphosphonates on acid phosphohydrolases and other lysosomal enzymes.
Diphosphonates are known to inhibit bone resorption in tissue culture and in experimental animals. This effect may be due to their ability to inhibit the dissolution of hydroxyapatite crystals, but other mechanisms may be important. Since lysosomal enzymes have implicated in the process of bone resorption, we have examined the effect of several phosphonates and of a polyphosphate (P20,2) on lysosomal hydrolases derived from rat liver and rat bone. Dichloromethylene diphosphonate strongly inhibited acid beta-glycerophosphatase (EC 3.1.3.2) and acid p-nitrophenyl phosphatase (EC 3.1.3.2) and to a lesser degree (in descending order) acid pyrophosphatase (EC 3.1.3.-), arylsulfatase A (EC 3.1.6.1), deoxyribonuclease II(EC 3.1.4.6) and phosphoprotein phosphatase (EC 3.1.3.16) of rat liver. Inhibition of acid p-nitrophenyl phosphatase and arylsulfatase A was competitive. Ethane-1-hydroxy-1, 1-diphosphonate did not inhibit any of these enzymes, except at high concentrations. Neither dichloromethylene diphosphonate nor ethane-1-hydroxy-1, 1-diphosphonate had any effect on beta-glucuronidase (EC 3.2.1.31), arylesterase (EC 3.1.1.2) and cathepsin D (EC 3.4.23.5). Of several other phosphonates tested only undec-10-ene-1-hydroxy-1, 1-diphosphonic acid inhibited acid p-nitrophenyl phosphatase strongly, the polyphosphate (P20, I) had little effect. Acid p-nitrophenyl phosphatase in rat calvaria extract behaved in the same way as the liver enzyme and was also strongly inhibited by dichloromethylene diphosphonate, but not by ethane-1-hydroxy-1, 1-diphosphonate. It is suggested that the inhibition of bone resorption by dichloromethylene diphosphonate might be due in part to a direct effect of this diphosphonate on lysosomal hydrolases.
Acid Phosphatase↗
Effect of diphosphonates on adenosine 3':5'-cyclic monophosphate in mouse calvaria after stimulation by parathyroid hormone in vitro.
1. The diphosphonates, disodium ethane-1-hydroxy-1,1-diphosphonate (EHDP) and disodium dichloromethylene diphosphonate (Cl2MDP), inhibit bone resorption in animals and in explanted bone in tissue culture. The possibility that these effects might be due to inhibition of skeletal adenylate cyclase has been studied. 2. EHDP and Cl2MDP, added for 30 min to the incubation medium at concentrations known to inhibit bone resorption, had no effect on basal content of adenosine 3':5'-cyclic monophosphate (cyclic AMP) of mouse calvaria incubated in vitro, nor did they inhibit the rise in cyclic AMP induced by bovine parathyroid hormone. 3. Pretreatment of mice for 3 days with Cl2MDP also had no effect on cyclic AMP under basal conditions or after incubation of explanted calvaria with parathyroid hormone in vitro. EHDP under similar conditions slightly inhibited the increase induced by parathyroid hormone but had no effect on basal concentrations of cyclic AMP. 4. It is suggested that the inhibition of adenylate cyclase is not an essential feature of the reduction of bone resorption by diphosphonates, which may act by direct inhibitory effects on the dissolution of hydroxyapatite and perhaps by other unidentified effects on bone cells. Key words: adenosine 3':5'-cyclic monophosphate, bone, dichloromethylene diphosphonate, diphosphonates, ethane-1-hydroxy-1,1-diphosphonate, parathyroid hormone.
Animals↗
The influence of orthophosphate on the renal handling of inorganic pyrophosphate in man and dog.
1. The urinary excretion of inorganic pyrophosphate (PPi), a known inhibitor of the growth and aggregation of crystals of calcium phosphate and calcium oxalate, increases after ingestion of orthophosphate (Pi). This effect may contribute to the apparent ability of oral phosphate to reduce the formation of urinary stones in man. This paper is a study of the mechanism by which Pi increases PPi excretion, investigated by renal clearance techniques in man and renal arterial infusion in dogs. PPi in plasma was measured by an isotope-dilution method after ion-exchange chromatography. 2. The mean renal clearance of endogenous PPi in ten men was 7-9 +/- 1-7 (SE) ml/min, and the mean ratio of PPi clearance to creatinine clearance was 0-08 +/- 0-02 (SE). The oral ingestion of Pi increased the urinary excretion and renal clearance of PPi about threefold, without significantly changing its concentration in plasma. 3. In dogs, the infusion of Pi into one renal artery caused a greater increase in urinary PPi from the infused than from the non-infused kidney, an effect that could be accentuated by simultaneous intravenous infusion of PPi. In dogs, only 1-3% of an injected or infused dose of PPi appeared intact in the urine, regardless of whether it was infused into the systemic or renal circulation. 4. These results suggest that Pi has a direct affect on the kidney to increase the excretion of PPi. It is possible that Pi either interferes with tubular reabsorption of PPi, perhaps by competing for a common tubular transport mechanism, or that Pi diminishes the intrarenal hydrolysis of PPi.
Animals↗
Inorganic phosphate homeostasis. Renal adaptation to the dietary intake in intact and thyroparathyroidectomized rats.
The possibility of renal tubular adaptation to variations in dietary inorganic phosphate (Pi) was investigated in intact and thyroparathyroidectomized (TPTX) rats pair-fed diets containing low, normal, and high amounts of Pi for periods up to 10 days. Clearances were measured before and during active i.v. infusions with Pi in conscious animals. Thus tubular reabsorption of phosphate (TRPi) could be assessed over a wide range of plasma phosphate concentrations ([Pi]P1). It was found that the renal tubule could adapt its capacity to transport Pi according to the dietary Pi: TRPi was always higher, for a given [Pi]P1, in the animals fed low than in those fed higher Pi diets. This diet-induced modification also occurred in the absence of thyroparathyroid glands, in the presence of the same calcemia and urinary pH, and during marked extracellular volume expansion. A time-course study in rats TPTX both before and during the administration of the experimental diets showed that a difference in the tubular handling of Pi was detectable as early as 3 days after switching the animals from a normal to low- or high-Pi diets. These results indicate that factors other than parathyroid hormone are implicated in the tubular response to variations in the dietary intake of inorganic phosphate.
Adaptation, Physiological↗