Category Archives: Minerals

Zinc-Carnosine > Zinc at Increasing Alkaline Phosphatase in Rat Tissue

Abstract

Comparison of the effect of beta-alanyl-L-histidinato zinc and its zinc-chelating ligand on bone metabolism in tissue culture.

The present investigation was undertaken to compare the effects of beta-alanyl-L-histidinato zinc (AHZ) and its zinc-chelating ligands on bone metabolism in tissue culture. Calvaria were removed from 3-week-old male rats and cultured for up to 72 h in Dulbecco’s modified eagle medium containing zinc sulfate, AHZ, di(N-acetyl-beta-alanyl-L-histidinato)zinc (AAHZ), and di(histidino)zinc (HZ). The bone calcium content and alkaline phosphatase activity were significantly increased in the presence of AHZ or AAHZ (10(-8)-10(-5) M). Those increases were seen at 10(-7) to 10(-5) M zinc sulfate and HZ. The bone deoxyribonucleic acid (DNA) content was significantly increased by AHZ or AAHZ with 10(-7) to 10(-5) M, while 10(-7) M zinc sulfate and HZ had no effect. Thus, AHZ and AAHZ had more potent effect than that of zinc sulfate and HZ. The effect of AHZ, AAHZ and HZ (10(-5) M) increasing bone alkaline phosphatase activity was abolished by the presence of 10(-4) M dipicolinate, a chelator of zinc. Moreover, the effect of these zinc compounds on bone metabolic indices was not seen in the presence of 10(-6) M cycloheximide. The present results suggest that the effect of AHZ and AAHZ on bone metabolism is more potent than that of zinc sulfate and HZ.

Yamaguchi M, Kishi S
Biol. Pharm. Bull. Apr 1994
PMID: 8069261

Zinc-Carnosine Stimulates Alkaline Phosphatase in Rat Bone Cells

Abstract

Stimulatory effect of beta-alanyl-L-histidinato zinc on alkaline phosphatase activity in bone tissues from elderly rats: comparison with zinc sulfate action.

The capability of beta-alanyl-L-histidinato zinc (AHZ) to increase alkaline phosphatase activity in the femoral diaphysis from elderly rats was investigated. The femoral-diaphyseal tissues were removed from weanling (3-week-old) and elderly (10-month-old) female rats. Bone tissues were cultured in Dulbecco’s modified Eagle medium (high glucose, 4.5%) supplemented with antibiotics and bovine serum albumin. Among various other bone-stimulating factors (AHZ; 10(-5) M, zinc sulfate; 10(-4) M, sodium fluoride; 10(-3) M, insulin; 10(-8) M, and beta-estradiol; 10(-9) M), AHZ had a potent effect on increasing alkaline phosphatase activity in the diaphyseal tissues from both rat groups. In the bone tissues from elderly rats, the effect was concentration dependent (10(-7) – 10(-5) M). At 10(-5) M the effect of AHZ was seen for a longer time during 72-h culture, although the zinc sulfate (10(-5) M) effect was no longer. The effect of AHZ to increase bone alkaline phosphatase activity was completely abolished by the presence of cycloheximide (10(-6) M). AHZ thus appears able to directly stimulate alkaline phosphatase activity dependent on protein synthesis in the bone tissues from elderly rats.

Kisi S, Yamaguchi M
Biol. Pharm. Bull. Feb 1994
PMID: 8205136

Zinc-Carnosine Can Prevent Bone Loss in Ovariectomized Rats

Abstract

Histomorphological confirmation of the preventive effect of beta-alanyl-L-histidinato zinc on bone loss in ovariectomized rats.

The effect of beta-alanyl-L-histidinato zinc (AHZ), in which zinc is chelated to beta-alanyl-L-histidine, on bone loss was investigated in the femur of ovariectomized rats. AHZ (10, 30 and 100 mg/kg/d) was orally administered to ovariectomized rats for 3 months. Ovariectomy significantly decreased the estradiol concentration in the serum as compared with that from sham-operated rats. This decrease was not altered by the dose of AHZ. The bone ash weight and mineral density in the femur of ovariectomized rats significantly decreased in comparison with those from sham-operated rats. Moreover, the trabecular bone at the femoral metaphysis was clearly decreased by ovariectomy. The decreases in the femoral ash content and mineral density and the metaphyseal trabecular bone were clearly prevented by the tested doses of AHZ (10, 30 and 100 mg/kg/d). The present finding with histomorphological study further supports the view that the administration of AHZ can prevent bone loss by ovariectomy.

Kishi S, Segawa Y, Yamaguchi M
Biol. Pharm. Bull. Jun 1994
PMID: 7951157

Zinc-Carnosine Prevents Bone Loss in Ovariectomized Rats

Abstract

Prolonged administration of beta-alanyl-L-histidinato zinc prevents bone loss in ovariectomized rats.

The effect of beta-alanyl-L-histidinato zinc (AHZ), in which zinc is chelated to beta-alanyl-L-histidine, on bone metabolism was investigated in the femoral diaphysis of ovariectomized rats. AHZ (10, 30 and 100 mg/kg body weight/day) was orally administered to ovariectomized rats for 3 months. Ovariectomy significantly decreased the estradiol concentration in the serum as compared with that from sham-operated rats. This decrease was not altered by the dose of AHZ. The bone volume and dry weight in the femur of ovariectomized rats significantly decreased in comparison with those from sham-operated rats. Moreover, alkaline phosphatase activity, deoxyribonucleic acid (DNA) and calcium contents in the femoral diaphysis were significantly decreased by ovariectomy. The decreases of the femoral volume and dry weight, the femoral-diaphyseal alkaline phosphatase activity, DNA and calcium contents by ovariectomy were completely prevented by the tested doses of AHZ (10, 30 and 100 mg/kg/day). AHZ in the dose range of 10-100 mg/kg/day caused a significant increase in zinc content in the femoral diaphysis of ovariectomized rats. The present study suggests that the prolonged administration of AHZ can prevent bone loss by ovariectomy.

Yamaguchi M, Kishi S
Jpn. J. Pharmacol. Oct 1993
PMID: 8283831 | Free Full Text

Zinc Restores Protein Synthesis in Unloaded Rats

Abstract

Zinc stimulates protein synthesis in the femoral-metaphyseal tissues of normal and skeletally unloaded rats.

The effect of zinc on protein synthesis in the femoral-metaphyseal tissues of normal and skeletally unloaded rats was investigated. Skeletal unloading was designed using the model of hindlimb suspension in rats. Animals were fed for 2 or 4 days during the unloading. [3H]Leucine was added to the reaction mixture containing the 5500 g supernatant fraction of the homogenate prepared from the femoral-metaphyseal tissues. In vitro protein synthesis was significantly decreased in the bone tissues from the rats which had undergone unloading for 2 or 4 days. When the metaphyseal tissues were cultured for 24 h in the presence of zinc sulfate (10(-5) M) or beta-alanyl-L-histidinato zinc (AHZ, 10(-5) M), zinc compounds clearly stimulated protein synthesis in the metaphyseal tissues from the 4-day unloaded rats. The zinc effect was also seen in the metaphyseal tissues from normal rats. The addition of zinc sulfate (10(-5) M) or AHZ (10(-7) to 10(-5) M) into the reaction mixture containing the 5500 g supernatant fraction of metaphyseal homogenate from normal or unloaded rats produced a significant increase in protein synthesis. This increase was clearly inhibited in the presence of cycloheximide (10(-7) M). The present result demonstrates that protein synthesis is impaired in the femoral-metaphyseal tissues of rats with skeletal unloading, and that this impairment is clearly restored by zinc supplementation.

Ehara Y, Yamaguchi M
Res Exp Med (Berl) 1997
PMID: 9089885

Zinc-Carnosine Has Proliferative Effect on Mouse Osteoblasts In Vitro

Abstract

Stimulatory effect of beta-alanyl-L-histidinato zinc on cell proliferation is dependent on protein synthesis in osteoblastic MC3T3-E1 cells.

The effect of beta-alanyl-L-histidinato zinc (AHZ) on bone metabolism was investigated in osteoblastic MC3T3-E1 cells. Cells were cultured for 3 days at 37 degrees C in a CO2 incubator in plastic dishes containing alpha-modified minimum essential medium supplemented with 10% fetal bovine serum. After the cultures, the medium was exchanged for that containing 0.1% bovine serum albumin plus various concentrations of AHZ or other reagents, and the cells were cultured further for appropriate periods of time. The presence of AHZ (10(-7)-10(-5) M) stimulated the proliferation of cells. AHZ (10(-6) and 10(-5) M) increased deoxyribonucleic acid (DNA) content in the cells with 48 hr-culture. This increase was completely blocked by the presence of cycloheximide (10(-6) M) or hydroxyurea (10(-3) M). Also, the presence of cycloheximide (10(-6) M) completely inhibited the AHZ (10(-5) M)-induced increase in the proliferation of cells. Meanwhile, parathyroid hormone (10(-7) M), estrogen (10(-9) M) and insulin (10(-8) M) significantly increased cellular DNA content. However, these hormonal effects clearly lowered in comparison with that of AHZ (10(-5) M). Dibutyryl cyclic AMP (10(-4) M) and zinc sulfate (10(-5) M) did not cause a significant increase in cellular DNA content. The present results support the view that AHZ has a direct specific proliferative effect on osteoblastic cells in vitro and that this effect is dependent on protein synthesis.

Hashizume M, Yamaguchi M
Mol. Cell. Biochem. May 1993
PMID: 8350864

Zinc-Carnosine > Zine at Stimulating DNS Synthesis in Mouse Osteoblast Cells

Abstract

Stimulatory effect of zinc-chelating dipeptide on deoxyribonucleic acid synthesis in osteoblastic MC3T3-E1 cells.

Whether deoxyribonucleic acid (DNA) synthesis in osteoblastic MC3T3-E1 cells is stimulated by zinc, an activator of bone formation, was investigated in vitro. After subculture for 3 days, the cells were cultured for up to 3 days (72 h) with zinc sulfate or zinc-chelated dipeptide (beta-alanyl-L-histidinato zinc; AHZ) in the range of 10(-7) to 10(-5) M. The culture with zinc compounds (10(-5) M) produced a significant increase of cell number, DNA content, and protein concentration in the cells, as reported previously. The culture with zinc compounds (10(-6) and 10(-5) M) clearly stimulated DNA synthesis in the homogenate, when it was estimated by the incorporation of [3H]deoxythymidine 5′-triphosphate into the DNA in the homogenate of cells. The AHZ effect was greater than that of zinc sulfate. The culture together with cycloheximide (19(-6) M) completely abolished the zinc compounds (10(-5) M)-induced increase of DNA synthesis in the cells, suggesting that the zinc compound effect is based on a newly synthesized protein component. Moreover, when zinc sulfate (10(-7) and 10(-6) M) or AHZ (10(-8) to 10(-5) M) was added into the reaction mixture with the homogenate of cells cultured without zinc compounds, the DNA synthesis was clearly increased. The effect of addition of zinc compounds (10(-6) M) on the DNA synthesis was completely inhibited by the presence of staurosporine (10(-8) M), an inhibitor of protein kinase C, or okadaic acid (10(-7) M), an inhibitor of protein phosphatase. The present study demonstrates that zinc compounds have a stimulatory effect on DNA synthesis in osteoblastic cells.

Yamaguchi M, Matsui T
Peptides 1996
PMID: 8959758

 

Zinc-Carnosine Prevents Deterioration of Bone in Ovariectomized Rats

Abstract

Preventive effect of beta-alanyl-L-histidinato zinc on the deterioration of bone metabolism in ovariectomized rats.

The preventive effect of beta-alanyl-L-histidinato zinc (AHZ) on the deterioration of bone metabolism was investigated in the femoral diaphysis of ovariectomized rats. AHZ (10, 30 and 100 mg/kg body weight/d) was orally administered to ovariectomized rats for 6 weeks. Ovariectomy produced a significant decrease in estradiol, calcitonin, calcium and inorganic phosphorus concentrations in the serum as compared with those from sham-operated rats. The dose of 30 and 100 mg AHZ/kg prevented any decrease in serum inorganic phosphorus concentration caused by ovariectomy. Alkaline phosphatase activity, deoxyribonucleic acid (DNA) and calcium contents in the femoral diaphysis of ovariectomized rats significantly decreased in comparison with those from sham-operated rats. These decreases were completely prevented by the dose of AHZ (10, 30 and 100 mg/kg). Electron microscopical analysis showed a rough alteration of bone matrix in the femoral diaphysis of ovariectomized rats. This alteration was clearly modified by the doses of AHZ (10, 30 and 100 mg/kg). Also, dosages of AHZ (30 and 100 mg/kg) restored the atrophy of osteoblasts and cartilage cells caused by ovariectomy. The present study suggests that oral administration of AHZ can prevent the deterioration of bone metabolism by ovariectomy. AHZ may have a therapeutic role in the treatment of osteoporosis.

Segawa Y, Tsuzuike N, Tagashira E, Yamaguchi M
Biol. Pharm. Bull. May 1993
PMID: 8364496

Zinc Inhibits Osteoclast-Like Cell In Rat Cells

Abstract

Zinc compounds inhibit osteoclast-like cell formation at the earlier stage of rat marrow culture but not osteoclast function.

The effect of zinc compounds on osteoclast-like cell formation in rat marrow culture in vitro was investigated. The bone marrow cells were cultured for 7 days in alpha-minimal essential medium containing a well-known bone resorbing hormone (1, 25-dihydroxyvitamin D3 and parathyroid hormone [1-34]). Osteoclast-like cell formation was estimated by staining for tartrate-resistant acid phosphatase (TRACP), a marker enzyme of osteoclasts. The presence of 1, 25-dihydroxyvitamin D3 (10(-8) M) or parathyroid hormone (PTH; 10(-8) M) induced a remarkable increase in osteoclast-like multinucleated cells (MNC). These increases were clearly inhibited by the presence of zinc sulfate or zinc-chelating dipeptide (beta-alanyl-L-histidinato zinc; AHZ) in the concentration range of 10(-7) to 10(-5) M. The inhibitory effect was seen at the earlier stage of osteoclast-like MNC formation. However, zinc compounds (10(-6) M) did not have an effect on PTH (10(-8) M)-induced osteoclast-like cell formation in the presence of EGTA (5 x 10(-4) M), dibucaine (10(-5) M) or staurosporine (10(-9) M). Moreover, when osteoclasts isolated from rat femoral-diaphyseal tissues were cultured for 24 h in the presence of zinc compounds (10(-7) to 10(-5) M), the compounds did not have an effect on cell numbers or lysosomal enzymes activity (acid phosphatase and beta-glucuronidase) in the cells. The present study clearly demonstrates that zinc compounds inhibit osteoclast-like cell formation at the earlier stage with differentiation of marrow cells.

Yamaguchi M, Kishi S
Mol. Cell. Biochem. May 1996
PMID: 8817479

Zinc-Carnosine Stimulates Bone Formation in Arthritic Rats

Abstract

Effect of beta-alanyl-L-histidinato zinc on bone metabolism in rats with adjuvant arthritis.

The effect of a new zinc compound, beta-alanyl-L-histidinato zinc (AHZ), on osteopenia was investigated in rats with adjuvant arthritis. Arthritis was induced in female rats by administering 1% Mycobacterium butyricum (MB) into the subplantar surface of the right hind paw. AHZ (10, 30 and 100 mg/kg body weight) was orally administered to MB-treated rats 28 times at 24-h intervals, and the rats were bled 24 h after the last administration. Treatment with MB caused a remarkable increase in paw volume and a corresponding decrease in the ratio of albumin per globulin in serum, indicating that the treatment induces inflammation. These alterations were not significantly changed by the administration of AHZ (10, 30 and 100 mg/kg). Serum calcium and zinc concentrations are significantly decreased in rats with adjuvant arthritis. These decreases were completely restored by the administration of AHZ (30 and 100 mg/kg). Furthermore, the inflammation-induced decreases in alkaline phosphatase activity and calcium content in the femoral diaphysis were clearly blocked by the administration of AHZ (30 and 100 mg/kg). Also, the larger doses of AHZ (30 and 100 mg/kg) produced a significant increase in femoral-diaphyseal deoxyribonucleic acid and in the zinc content in rats with adjuvant arthritis. These results suggest that AHZ has a stimulating effect on bone formation in the femoral diaphysis of rats with adjuvant arthritis, although the compound did not have an anti-arthritic effect.

Segawa Y, Tsuzuike N, Itokazu Y, Tagashira E…
Biol. Pharm. Bull. Jul 1993
PMID: 8401397