acid-phosphatase and ipriflavone

To investigate the therapeutic effects of ipriflavone on postmenopausal syndrome and osteoporosis in women.. A randomized and double-blind study was conducted. Sixty postmenopausal women with osteoporosis were chosen and they were randomly divided into three groups: Treatment group I was given oral compound calcium acid chelate and Vitamin AD guttate; treatment group II was given oral compound calcium acid chelate, Vitamin AD guttate and ipriflavone; Control group was given placebo and compound calcium acid chelate. The postmenopausal syndrome, bone mineral density (BMD), and bone biochemical markers were assessed 6 and 12 months after the treatment.. In treatment group II, hot flush and ostalgia syndromes were dramatically relieved, BMD and serum calcium level increased markedly and alkaline phosphatase, parathyroid hormone and tartrate-resistant acid phosphatase decreased markedly, comparing with treatment group I and control group (p < 0.05).. Ipriflavone could inhibit bone resorption and promote bone formation. It is an effective drug for the prevention and treatment to menopausal syndrome and osteoporosis. Ipriflavone could be used as a supplement to estrogen replacement treatment.

Topics: Acid Phosphatase; Adult; Alanine Transaminase; Bone Density; Bone Remodeling; Calcium; Double-Blind Method; Female; Humans; Isoenzymes; Isoflavones; Menopause; Osteoporosis, Postmenopausal; Parathyroid Hormone; Phosphorus; Phytoestrogens; Statistics, Nonparametric; Tartrate-Resistant Acid Phosphatase

Thirty postmenopausal women with reduced bone mineral density were divided randomly into two groups based on the chronological sequence of their first visit to the Osteoporosis Clinic of Katsuragi Hospital. Group I was given 600 mg ipriflavone orally daily and group II was weekly injected intramuscularly with 20 units elcatonin, Asu1-7 eel calcitonin (carbocalcitonin). Lumbar spine BMD was measured by dual-energy X-ray absorptiometry, and trabecular bone mineral density at the distal radius, cortical bone density, and relative cortical volume at the radial diaphysis by peripheral computed tomography before the beginning of the study and at the 4th, 8th, and 12th month. Markers of bone metabolism--serum total alkaline phosphatase, bone-specific alkaline phosphatase, tartrate-resistant acid phosphatase, osteocalcin, intact osteocalcin, PICP and ICTP, and urinary pyridinoline, deoxypyridinoline, and calcium/creatinine (Ca/Cr)--were also measured at the same interval. Plasma parathyroid hormone (PTH) and calcitonin (CT) were measured at the same time. Radial trabecular bone density showed a significantly higher rate of increase in group I (ipriflavone group) than in group II (elcatonin group) at the 4th month, whereas lumbar spine BMD showed more pronounced increase in the elcatonin group than in the ipriflavone group throughout the study period. Bone metabolism markers tended to decline in both groups. Total and intact osteocalcin showed a significant fall from the baseline throughout the study period only in the ipriflavone group. Urine pyridinoline and deoxypyridinoline showed a significant fall from the baseline at the 12th month only in the ipriflavone group. On comparing bone gainers with increase of lumbar spine BMD by 2% or more with bone losers with a decrease by 2% or more, only urine Ca/Cr was significantly different, lower in the former than in the latter, despite the general tendency for bone resorption markers to decrease in bone gainers and to increase in bone losers.

Topics: Acid Phosphatase; Aged; Alkaline Phosphatase; Animals; Bone Remodeling; Bone Resorption; Calcitonin; Eels; Female; Humans; Isoenzymes; Isoflavones; Middle Aged; Osteocalcin; Parathyroid Hormone; Tartrate-Resistant Acid Phosphatase

Ipriflavone (7-isopropoxyisoflavone) is an effective antiresorptive agent used to treat osteoporosis. However, the mechanism of its action on osteoclasts and their precursor cells is not well understood. To determine whether the mechanism involves direct effects on osteoclasts or their precursors, we examined the effects of ipriflavone on cytosolic free calcium ([Ca2+]i) in osteoclasts and their precursors and measured specific binding of 3H-labeled ipriflavone. Highly purified chicken osteoclast precursors, which spontaneously differentiate into multinucleated osteoclasts in 3-6 days, were loaded with fura-2, and the subcellular [Ca2+]i distribution was monitored by videoimaging. Ipriflavone induced a rapid increase in [Ca2+]i followed by a sustained elevation [EC50 = 5 x 10(-7) M, 263 +/- 74 nM (SE) (n = 8) above basal levels, by 10(-6) M ipriflavone, sustained phase]. The responses were the same in differentiated chicken osteoclasts and isolated rabbit osteoclasts. An influx of extracellular Ca2+ is likely to be responsible for the ipriflavone-induced change in [Ca2+]i because the response was abolished by 0.5 mM LaCl3, or by Ca-free medium containing EGTA. Moreover, high [Ca2+]i levels were detected adjacent to the cell membrane after ipriflavone addition. Ipriflavone induced Ca influx mainly through dihydropyridine-insensitive Ca2+ channels, because nicardipine (10(-7)M) and verapamil (10(-7)M) had no effects on ipriflavone-induced [Ca2+]i responses. [3H]Ipriflavone binding studies indicated the presence of specific ipriflavone binding sites (two classes), both in precursor cells [dissociation constant (Kd), 7.60 x 10(-8)M, 2.67 x 10(-6)M] and in mature osteoclasts (Kd, 4.98 x 10(-8)M, 3.70 x 10(-6)M). Specific ipriflavone binding was not displaced by various modulators of avian osteoclast function, such as estradiol (10(-8)M) or retinoic acid (10(-6)M), indicating that ipriflavone receptors differ from the receptors for these Ca-regulating hormones. The fusion of osteoclast precursor cells was significantly inhibited by ipriflavone, which led to dose-dependent inhibition of bone resorption and tartrate-resistant acid phosphatase activity. Novel specific ipriflavone receptors that are coupled to Ca2+ influx were demonstrated in osteoclasts and their precursor cells. These ipriflavone receptors may provide a mechanism to regulate osteoclast differentiation and function.

Topics: Acid Phosphatase; Animals; Bone Remodeling; Bone Resorption; Calcium; Cell Division; Chickens; Cytosol; Female; Fluorescent Dyes; Fura-2; Isoenzymes; Isoflavones; Osmolar Concentration; Osteoclasts; Rabbits; Receptors, Vitronectin; Tartrate-Resistant Acid Phosphatase

The effects of ipriflavone and vitamin K on bone metabolism were examined using a culture system. Vitamin K1 and vitamin K2 (10(-7)M-10(-5)M) inhibited both the activation of mature osteoclasts and the formation of new osteoclasts without affecting the growth of progenitor cells in cultures of mouse unfractionated bone cells. The inhibitory effects of vitamin K on bone resorption were similar to those of ipriflavone and were not affected by the vitamin K antagonist warfarin. When ipriflavone was added to the culture medium in combination with vitamin K2, an additive inhibitory effect on bone resorption was observed. An additive effect was also observed in organ cultures of mouse calvaria. On the other hand, ipriflavone, but neither vitamin K1 nor vitamin K2, stimulated cellular alkaline phosphatase (ALP) activity on rat bone marrow stromal cells under culture conditions in which cells subsequently form mineralized bone-like tissue. Vitamin K1 and vitamin K2 also did not modulate the stimulatory effect of ipriflavone on the ALP activity of the cells. These results suggest that the inhibitory effects of vitamin K on bone resorption are similar to those of ipriflavone through mechanisms that may be independent of the gamma-carboxylation system, while the effects of vitamin K on osteoblast phenotype expression are different from those of ipriflavone.

Topics: Acid Phosphatase; Animals; Bone Development; Bone Marrow; Bone Resorption; Cell Division; Cell Fractionation; Cells, Cultured; Drug Synergism; Femur; Isoflavones; Mice; Organ Culture Techniques; Osteoclasts; Phenotype; Rats; Stem Cells; Stromal Cells; Vitamin K; Warfarin

The effects of ipriflavone on bone cells were studied in vitro on pre-osteoclastic (FLG 29.1) and osteoblast-like (Saos-2) cells grown for 48 h either separately or in co-cultures, with or without the addition of PTH. Histological, ultrastructural and histochemical (TRAP-activity demonstration) methods were used. The main results show that ipriflavone reduces replication of FLG 29.1 cells and inhibits TRAP production by these cells both in controls and in co-cultures treated with PTH. Moreover, it has a moderate stimulatory effect on proliferation of osteoblast-like cells and reduces the PTH-induced degenerative changes of Saos-2 cells. These results suggest that the inhibitory effect of ipriflavone on FLG 29.1 cells might be indirect and might be mediated by the osteoblast-like cells.

Topics: Acid Phosphatase; Cell Adhesion; Cell Division; Filaggrin Proteins; Humans; Isoflavones; Microscopy, Electron; Osteoblasts; Osteoclasts; Parathyroid Hormone; Tumor Cells, Cultured

Bone labeling, histochemical, and fine structural studies were performed in order to clarify the effects of ipriflavone (IP) on rat bone tissue in vivo and in vitro. Labeling experiments showed a slight increase in bone formation during 3 days' administration. It was also noted that many osteoclasts detached from the bone surface at 1, 2, and 6 hours after administration in vivo. In addition, irregular localization of tartrate-resistant acid phosphatase (TRACPase) activity was observed in osteoclasts. Fine structurally, IP-treated osteoclasts exhibited irregularity in their ruffled borders, as reported in calcitonin administration, and many enlarged rough endoplasmic reticuli and vacuoles were observed. However, osteoclasts at 12 hours after administration, as well as the control, indicated recovery features from the effect of IP. Osteoblast proliferation and differentiation led to increasing alkaline phosphatase activity (ALPase) with time as well as the development of rough endoplasmic reticuli and Golgi apparatus with well-developed fine structure. These findings imply active synthesis of bone matrix. In our in vitro experiment, osteoclasts and osteoblasts displayed histochemical and fine structural characteristics similar to those observed in our in vivo experiment. Moreover, fewer TRACP-positive mononuclear cells were observed after 24-hour culture with IP than with the control. These results suggest that IP inhibits directly and/or indirectly differentiation and activity of osteoclasts and also promotes differentiation of osteoblast-lineage cells and their bone-forming activity.

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Bone Resorption; Cell Differentiation; Endoplasmic Reticulum; Golgi Apparatus; In Vitro Techniques; Isoenzymes; Isoflavones; Male; Osteoblasts; Osteoclasts; Osteogenesis; Rats; Rats, Wistar; Tartrate-Resistant Acid Phosphatase

Effects of ipriflavone (7-isopropoxyisoflavone) on osteoclast-induced bone resorption were evaluated using an unfractionated bone cell culture system containing mature osteoclasts from the femur and tibia of newborn mice. When cells were cultured for 4 days on dentin slices in the presence of 5% fetal bovine serum and 10(-8) M 1 alpha, 25(OH)2D3, ipriflavone (3 x 10(-7) -3 x 10(-5) M) inhibited pit formation and caused a decrease in the number of tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cells (MNCs). The lowest significant effect was observed at a concentration of 10(-6) M. Unlike ipriflavone, calcitonin inhibited pit formation 4 days after the culture was started without affecting the number of TRAP-positive MNCs. Ipriflavone still inhibited pit formation when the culture period was 13 days, when new osteoclasts were expected to be formed. These findings suggest that ipriflavone inhibits new osteoclast formation and bone resorption at the cellular level.

Topics: Acid Phosphatase; Animals; Bone Resorption; Calcitriol; Cells, Cultured; Dentin; Isoflavones; Mice; Osteoclasts

Ipriflavone, one of the isoflavone derivatives, is a therapeutic drug for osteoporosis. The mechanism is thought to be the inhibition of bone resorption. In the present paper, we report that ipriflavone inhibited formation of osteoclasts from murine spleen cells co-cultured with stromal cells cloned from murine bone marrow. In this system, ipriflavone inhibited osteoclast generation in a dose-dependent manner (10(-7)-10(-5) M). Ipriflavone also inhibited prostaglandin E2 production in MC3T3-E1 cells, which are widely employed as osteoblasts. Moreover, ipriflavone inhibited the proliferation of stromal cells (10(-6)-10(-5) M), but not osteoblastic cells. These results suggest that one mechanism for the inhibitory effects of ipriflavone on bone resorption is the inhibition of osteoclast formation through inhibiting prostaglandin E2 production in osteoblasts and thereby suppressing proliferation of stromal cells.

Topics: 3T3 Cells; Acid Phosphatase; Animals; Bone Marrow; Bone Marrow Cells; Cell Division; Cells, Cultured; Dinoprostone; Isoflavones; Male; Mice; Osteoblasts; Osteoclasts; Spleen; Stromal Cells

The effects of internal treatment were compared with those of surgical procedures on secondary hyperparathyroidism as a complication of chronic renal failure patients in order to study their applications and problems. Maintenance hemodialysis patients complicated with 2HPT were selected as test subjects. The following internal treatment was administered. 1) Elcitonine to 6 cases and 2) Ipriflavon to 5 cases respectively and applied 3) pulse therapy of 1.25(OH)2D3 to 12 cases. On the other hand, total parathyroidectomy and autotransplantation were carried out as surgical procedures. The results of the treatment were evaluated by comparing serological data including tartrate resistant acid phosphatase (TAP) measured with the passage of time, bone scintigram findings, and change of bone mineral content (BMC) measured by single photon absorptiometry and dual photon absorptiometry. In the pulse therapy group, an oral tolerance test of 6 micrograms of 1.25(OH)2D3 was carried out to investigate its relation to long term prognosis. Furthermore, in both the pulse therapy group and PTX group, serum aluminum (A1) and delta A1 calculated by Defferoxamine (DFO) tolerance test were measured. The results are as follows. 1) In Elcitonine and Ipriflavon administration groups, increase of ALP and PTH and decrease of BMC (p less than 0.05) were recognized. 2) In the pulse therapy group, although the patients with PTH-C less than 30 ng/ml showed decrease in PTH both in short and long terms, cases with PTH-C more than 30 ng/ml kept the same level in PTH. Regardless of the change of PTH and ALP, there was no significant change observed in BMC. 3) In the PTX group, ALP/TAP ratio rose by 900% temporarily and BMC increased (p less than 0.01) in all regions measured. 4) Serum A1 and delta A1 were decreased (p less than 0.01) in PTX cases and in the pulse therapy cases in which ALP was decreased. In recent years, internal treatments on 2HPT patients have become diversified. However, exacerbation of 2HPT considered as an escape phenomenon caused the decrease in BMC after exclusive calcitonine preparatives administration. Pulse therapy, which is regarded at present as most effective in reducing PTH, ALP, did not work to increase BMC efficiently. In other word, in order to attain efficient BMC increase action, it is necessary to transfer from bone resorption phase to formative phase rapidly with drastic decrease of PTH and TAP observed in PTX.(ABSTRACT TRUNCATED AT 400 WORDS)

Topics: Acid Phosphatase; Alkaline Phosphatase; Aluminum; Bone Density; Calcitonin; Calcitriol; Drug Administration Schedule; Drug Evaluation; Humans; Hyperparathyroidism, Secondary; Isoflavones; Kidney Failure, Chronic; Parathyroid Hormone