This study is to find out how the 4 types of calcium salt such as calcium carbonate, calcium phosphate, calcium lactate and calcium citrate in soy protein diet, the vegetable protein source, affect the calcium utilization in the body. To do so, calcium, phosphate and creatinine concentration and ALP activity in blood as well as the content of calcium and ash, the length, weight strength, and the calcium utilization in the bone were measured. Four groups of Sprague-Dawley male rats with the weight of around 180g were fed for 3 weeks with the experimental diet. Each group was fed with the isolated soy protein containing 14% of the diet and the above mentioned 4 types of calcium salt as the calcium source. The results are as follows; 1. There were no differences of the feed intake, weight gain, and feed efficiency among groups. 2. ALP activity in blood was sinificantly high in calcium lactate group(P<0.05), but there were no differences of concentration of calcium, phosphates, and creatinine in blood among groups. 3. The weight, calcium content, calcium ratio in ash and the strength of bone were low when calcium lactate was provided(P<0.05). 4. The content of calcium in the liver was high in calcium lactate group and calcium citrate group(P<.0.05). 5. The exceretion of feces was low in calcium lactate group(P<0.05) and the excretion of urine was also relatively low. In addition, the ratio of absorption and the retention of calcium were high(P<0.05). In summary, out of four types of calcium salt such as calcium carbontate, calcium phosphate, calcium lactate and calcium citrate when calcium lactate was provided the ALP activity in blood was high and the weight, calcium content, calcium ratio in ash and the strength of bone were low. In calcium utilization, the ratio of absorption and retention of calcium were high, however it has lower effect than 3 other calcium types in improving weight, the content of calcium and the strength of bone.
This study was performed to investigate the effects of dietary protein and calcium levels on calcium metabolism in eight healthy Korean adult females. The 2-day metabolic study consisted of a 2 day adaptation period and three 6-day experimental periods. Three experimental diets were low protein low calcium(LPLCa : protein 44g, Ca 422mg), higher protein low calcium(HPLCa : protein 85g, Ca 365mg), and high protein high calcium (HPHCa : protein 84g, Ca 727mg). The apparent calcium absorption was likely to be affected by the calcium intake rather than by the protein intake. Average calcium absorption rate was about 23-29% of calcium intake. The calcium balance was -21.44mg for LPCa, -25.02mg for HPLCa, and -3.22mg for HPHCa. Avergae urinary calcium excretion was 127.7mg for LPLCa, 108.6mg for HPLCa, and 215.4mg for HPHCa. Urinary calcium excretion was more closely related to the changes of calcium intake rather than of protein intake. These results seemed to be due to the interactions between the high phosphours contained in the high protein diet and the little discrepancy of protein intake levels.
Recently, we reported that high extracellular calcium increased receptor activator of nuclear factor-${\kappa}B$ ligand (RANKL) expression via p44/42 mitogen-activated protein kinase (p44/42 MAPK) activation in mouse osteoblasts. However, the mechanism for p44/42 MAPK activation by high extracellular calcium is unclear. In this study, we examined the role of intracellular calcium increase in high extracellular calcium-induced RANKL induction and p44/42 MAPK activation. Primary cultured mouse calvarial osteoblasts were used. RANKL expression was highly induced by 10 mM calcium treatment. Ionomycin, a calcium ionophore, also increased RANKL expression and activated p44/42 MAPK. U0126, an inhibitor of MEK1/2, an upstream activator of p44/42 MAPK, blocked the RANKL induction by both high extracellular calcium and ionomycin. High extracellular calcium increased the phosphorylation of proline-rich tyrosine kinase 2 (Pyk2), one of the known upstream regulators of p44/42 MAPK activation. Bisindolylmaleimide, an inhibitor of protein kinase C, did not block RANKL induction and p44/42 MAPK activation induced by high extracellular calcium. 2-Aminoethoxydiphenyl borate, an inhibitor of inositol 1,4,5-trisphosphate (IP3) receptor, blocked the RANKL induction by high extracellular calcium. It also partially suppressed the activation of Pyk2 and p44/42 MAPK. Cyclosporin A, an inhibitor of calcineurin, also inhibited high calcium-induced RANKL expression in dose dependent manner. However, cyclosporin A did not affect the activation of Pyk2 and p44/42 MAPK by high extracellular calcium treatment. These results suggest that 1) the increase in intracellular calcium via IP3-mediated calcium release is necessary for RANKL induction by high extracellular calcium treatment, 2) Pyk2 activation, but not protein kinase C, following the increase in intracellular calcium might be involved in p44/42 MAPK activation, and 3) calcineurin-NFAT activation by the increase in intracellular calcium is involved in RANKL induction by high extracellular calcium treatment.
Calcium is the most abundant stored mineral in the human body and is especially vital for bone health; thus, calcium deficiency can cause bone-related diseases, such as osteopenia and osteoporosis. However, a high concentration of serum calcium, which is commonly known as hypercalcemia, can also lead to weakened bones and, in severe cases, osteosarcoma. Therefore, it is necessary to maintain the concentration of calcium that is appropriate for bone biology. In the present study, we aimed to elucidate the effects of high concentration of calcium, approximately 2 folds the normal calcium level, on osteoblast differentiation. The CaCl2 treatment showed dose-dependent suppression of the alkaline phosphatase activity and mineralized nodule formation. Calcium showed cytotoxicity at an extremely high concentration, but a moderately high concentration of calcium that results in inhibitory effects to osteoblast differentiation showed no signs of cytotoxicity. We also confirmed that the CaCl2 treatment repressed the mRNA expression and protein abundance of various osteogenic genes and transcriptional factors. Considered together, these results indicate that a high concentration of calcium negatively regulates the osteoblast differentiation of C2C12 cells.
This study was done to evaluate the effect of dietary calcium level (a diet which met 100% or twice the calcium level in AIN-76 diet) on preventing bone loss in ovariectomized rats. Forty Sprauge-Dawley female rats(body weight 200$\pm$5g)were divided into two groups. One group were ovariecotomized (Ovx) while the others received sham operation(Sham). Thereafter, each rat group was further divided into normal calcium diet(0.52%) and high calcium diet(1.04%) subgroups. All rats were fed on experimental diet and deionized water ad libitum for 8 weeks. The total body, spine and femur bone mineral densities and bone mineral contents were measured by Dual Energy X-ray Absorptiometry, Eight weeks following operation, ovariectomized rats fed a high calcium diet had a significantly higher total bone mineral content, total bone calcium content, spine bone mineral density, spine bone mineral content and femur bone mineral content than ovariectomized rats fed control calcium diet. The correlation between dietary calcium intake level and spine bone mineral density were positive, but there was no correlation between dietary calcium intake and femur bone mineral density. The findings from the present study demonstrated that bone loss due to ovarian hormonal deficiency can be partially prevented by a high calcium diet. Futhermore, these findings support the strategy of the use of a high calcium diet in the prevention of estrogen depleted bone loss(postmenopausal osteoporosis)
This study explored the effects of dietary calcium levels and/or ovariectomy on bone formation, bone composition and calcium metabolism using female Sprague-Dawley weanling rats(mean body weight$\pm$SEM : 232.3$\pm$6.7g) as a model. Rats received high(1.5%) calcium diets for eight weeks during the growth period and were randomly assigned to ovariectomy and sham groups. The two groups were than each randomly divided into three sub-groups and fed 0.1%, 0.5% and 1.5% calcium diets for eight weeks after operation. The results indicate that body weight gain was higher in ovariectomy groups than sham groups regardless of dietary calcium levels and food intakes. Serum Ca concentration was decreased in low Ca groups after operation and serum P concentration increased in ovariectomy groups. Serum Ca concentration was decreased in low Ca groups after operation and serum P concentration increased in ovariectomy groups. Serum alkaline phosphatase activity was increased in ovariectomy groups and was not influenced by dietary calcium levels after operation. Urinary hydroxyproline decreased in high Ca intake groups regardless of whether rats had received an ovariectomy or not. The weight, length and breaking force of the femur were not significantly different in all groups. Ash, calcium, phosphate and magnesium contents in the femur and lumbar were not significantly different regardless of ovariectomy operation and dietary calcium levels. But high/normal calcium intake after ovariectomy and sham operation increased the weight and calcium content in bone. Therefore, high calcium intake influenced the formation of peak bone mass during the growth period and calcium levels and calcium levels and calcium levels continued to influence bone growth and composition after ovariectomy.
This study was performed to investigate effect of dietary protein and calcium levels on cad-mium intoxication in rats. Adult Sprague-Dawley male rate(245$\pm$21g) were blocked into 18 groups of 7 animals according to body weight Nine experimental diets different with protein(40%, 15%, 7%) and calcium (1.3%, 0.6%, 0.1%) levels were prepared. Nine groups of animals were fed each diet with 50ppm cadmium in drinking water and the other 9 groups without cadmium for 30days. Results were summarized as follows: 1) Body weight gain F. E. R(Food Efficiency Ratio) and weights of liver kidney and femur were higher in high protein groups among cadmium exposed groups. 2) Cadmium contents in liver and intestine were higher in rats fed high protein diet or low calcium diet among cadmium exposed groups. Fecal cadmium excretion was highest in high protein-high calcium diet group among cadmium exposed animals. Metallothionein contents in liver kidney and intestine were higher in animals exposed to cadmium and fed high protein diets. 3) Gel filtration chromatography of cytosolic solution showed that the higher dietary protein and calcium levels were the more cadmium was found in metallothionein fractions. 4) No gross histopathological change was seen in liver kidney and intestine of cadmium exposed rats. However a significant increase of smooth endoplasmic reticulum which was alleveated by high protein-high calcium diet was observed. Results obtained indicated that not only high protein diet but also high calcium diet showed preventive effect on cadmium intoxication by increasing the induction of metallothionein syn-thesis and decreasing the cadmium absorption.
The purpose of this study was to investigate the effect of calcium intake on lipid contents and enzyme activities in rats of different ages. Lipid levels in serum and liver and GOT, CPK and LDH activities in serum were compared in rats of different ages(4 weeks and 10 months) that were fed various levels of calcium(50, 100, 200% of requirement)for 3 weeks. Body weight gain and feed efficiency ratio were significantly higher in young rats than in adults. Serum calcium level was increased by elevation of calcium intake levels were decreased. Liver phospholipid and triglyceride levels in the high-cal-terol and triglyceride levels were decreased. Liver phospholipid and triglyceride levels in the high calcium group were significantly lower than those in other groups. Serum GOT and LDH activities of adults were significantly higher in low-calcium group than those in adequate/high-calcium groups. However, serum CPK activity of adults was significantly higher in high-calcium group than that in low/adequate-calcium groups. The results of this study suggest that adequate calcium intake may have protective effects ont he alteration of lipid and enzyme activity in rats.
This study was done to evaluate the effectiveness of dietary calcium level(a diet which met 100% or twice the calcium level in AIN-76 diet) on preventing bone loss in ovariectomized rats. Forty female Sprauge-Dawley rats(body weight 200$\pm$5g) were divided into two groups. One group were ovariectomized(Ovx) while the others received sham operation(Sham). Thereafter, each rat group was further divided into normal calcium diet(NCD, 0.52%) and high calcium diet(HCD, 1.04%) sub-groups. All rats were fed on experimental diet and deionized water ad libitum for 8 weeks. Urinary pyridinoline & creatinine and serum estradiol, luteinizing hormone, calcium, phosphate, total protein, albumin, alkaline phosphatase and osteocalcin were determined. There were no significant differences in serum calcium. total protein and albumin in the two groups(Ovx vs Sham) of rats. Ovariectomized rats had significantly lower estradiol than sham operated rats. There was a highly significant correlation between total bone mineral density(TBMD) and overall level of esteradiol(r=0.59, p<0.05). Total bone mineral density did not correlate significantly with ALP or osteocalcin, although a negative trend was evident. However, the rats fed high calcium diet had a lower crosslinks value and osteocalcine than the rats fed normal calcium diet. An increased rate of bone turnover is usually associated with a decrease in bone mass bexause bone formation at each remodeling site is never as great as resorption. Ovariectomized rats fed high calcium diet had a lower crosslink value and osteocalcin; it means high cacium diet decreased bone turnover rate. The findings from the present study demonstrated that bone loss due to ovarian hormonal deficiency can be partially prevented by a high calcium diet. Futhermore, these findings support the strategy of the use of a high calcium diet in the prevention of estrogen depletion bone loss (postmenopausal osteoporosis).
To investigate the effects of dietary protein and calcium levels on calcium and bone metabolism Sprague-Dawley male growing rats weighting approximately 91.4g were divided into four groups and fed one of the following four experimental diets-15% protein 0.2% calcium ; 15% protein 0.5% calcium ; 30% protein 0.2% calcium ; 30% protein 0.5% calcium-for five weeks. Calcium intake and excretion, apparent calcium absorption were measured and bone densities and mineral contents of femur and scapula were analyzed. Calcium excretion through feces and urine was significantly greater in animals receiving diets of higher calcium. Fecal calcium but not urinary calcium excretion was greater when the protein level was increased from 15% to 30%. Apparent calcium absorption rate was significantly higher with lower calcium intakes. Serum alkaline phosphatase activity was significantly higher in 0.2% calcium group than in 0.5% calcium group, while urinary hydroxyproline excretion was essentially same among all experimental groups. Weights and mineral contents or protein. Bone weights were greater, but calcium and ash contents of femur and scapula were lower in animals on the diet containing low calcium and high protein, which suggests that bone metabolism may be affected by the interaction between calcium and protein intake. These results indicate that during growth high protein intake might be beneficial to bone health if the diet is sufficient in calcium, however, if the diet fails to provide an optimum amount of calcium, such practice might be detrimental.
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