acid-phosphatase and Ascorbic-Acid-Deficiency

The transcription factor, nuclear factor-kappa B (NFkappaB), is believed to play a pivotal role in osteoclast formation. In this study, we focused on NFkappaB decoy oligodeoxynucleotides (ODN) as a new therapeutic strategy to attenuate osteoporosis. Tartrate-resistant acid phosphatase (TRAP)-positive multinuclear osteoclasts formed in mononuclear cells including osteoclast precursors from neonatal rabbit bone marrow were increased in the presence of 1,25-dihydroxyvitamin D3, whereas transfection of NFkappaB decoy ODN decreased the number of TRAP-positive cells and attenuated RANKL and M-CSF-induced osteoclast formation. NFkappaB decoy ODN also inhibited the activity of osteoclasts, as assessed by pit formation. In rat ovariectomized model of estrogen deficiency, continuous administration of NFkappaB decoy ODN attenuated the increase of TRAP activity, accompanied by a significant increase in calcium concentration in tibia and femur and decrease in urinary deoxypyridinoline. In additional osteoporosis model using vitamin C-deficient rat, inhibition of NFkappaB by decoy ODN dramatically improved the bone length, weight, density as assessed by dual-energy X-ray absorptiometry. Overall, inhibition of NFkappaB by decoy strategy prevented osteoporosis through the inhibition of bone resorption. Targeting of NFkappaB might be potential therapy in various bone metabolic diseases.

Topics: Absorptiometry, Photon; Acid Phosphatase; Animals; Ascorbic Acid Deficiency; Cell Differentiation; Female; Femur; Genetic Therapy; Isoenzymes; Models, Animal; NF-kappa B; Oligodeoxyribonucleotides; Osteoclasts; Osteoporosis; Ovariectomy; Rabbits; Rats; Rats, Wistar; Tartrate-Resistant Acid Phosphatase; Transfection

The effect of ascorbic acid deficiency on bone metabolism was evaluated using the ascorbate-requiring Osteogenic Disorder Shionogi (ODS) rat model. Ascorbic acid (Asc)-deficient rats gained body weight in a manner similar to Asc-supplemented rats (control) during 3 weeks, but began to lose weight during the 4th week of Asc deficiency. The tartrate-resistant acid phosphatase (TRAP) activity in serum increased to about 2-fold the control value in the rats fed the Asc-free diet for 2, 3, and 4 weeks (AscD2, AscD3, and AscD4), while a decrease in the alkaline phosphatase (ALP) activity was observed only in AscD4 rats. The serum pyridinoline cross-linked carboxyterminal telopeptide of type I collagen (ICTP) level significantly increased to 1.3-, 1.4-, and 1.9-fold of that in the controls in AscD2, D3, and D4, respectively. The ALP activity in the distal femur was unchanged in AscD1, D2, and D3, but decreased to 50% of the control level in AscD4 rats. The TRAP activity in the distal femur increased to about 2-fold of that in the controls in the AscD2 and D3 and decreased to the control level in the AscD4 rats. The amount of hydroxyproline in the distal femur significantly decreased to about 80%, 70%, and 60% of the control in AscD2, D3, and D4 rats, respectively. These decreases were associated with a similar reduction in the calcium content of the distal femur. Histochemical analysis of the distal femur showed an increase in TRAP-positive cells in AscD2 and AscD3 rats and a decrease in the trabecular bone in AscD2, D3, and D4 rats. These results suggested that a deficiency of Asc stimulated bone resorption at an early stage, followed by a decrease in bone formation in mature ODS rats which already had a well-developed collagen matrix and fully differentiated osteoblasts.

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Ascorbic Acid Deficiency; Body Weight; Bone Resorption; Calcium; Collagen Type I; Disease Models, Animal; Female; Femur; Hydroxyproline; Immunohistochemistry; Isoenzymes; Oligopeptides; Peptide Fragments; Peptides; Procollagen; Rats; Tartrate-Resistant Acid Phosphatase; Time Factors

The effect of erythorbic acid (ErA) administration on activities of liver aniline hydroxylase, liver acid phosphatase, and serum alkaline phosphatase, and the content of liver cytochrome P-450 was studied to determine whether or not ErA would affect the availability of ascorbic acid (AsA) in normal and AsA-deficient guinea pigs. In experiment I, changes of the enzyme activities and liver cytochrome P-450 content in the guinea pigs administered AsA and/or ErA and sacrificed on days 4, 10, 16, and 30 were examined. Moreover, in experiment II, after 16 days of depletion of AsA, the guinea pigs were administered AsA and/or ErA. These animals were sacrificed on days 0, 4, and 20 of the repletion period, after which the activities of drug metabolic enzyme and phosphatases and content of cytochrome P-450 during recovery were observed. The enzyme activities and cytochrome P-450 content of AsA-supplemented guinea pigs were similar to those of ErA-supplemented animals and also similar to those of both AsA and ErA-supplemented guinea pigs throughout the experimental period. During the repletion of the AsA-depleted guinea pigs, there were no significant differences in these enzyme activities and cytochrome P-450 content among the animals administered AsA and/or ErA. These results suggested that ErA administration may not affect the AsA availability in the guinea pigs.

Topics: Acid Phosphatase; Alkaline Phosphatase; Aniline Hydroxylase; Animals; Ascorbic Acid; Ascorbic Acid Deficiency; Cytochrome P-450 Enzyme System; Guinea Pigs; Liver; Male

Topics: Acid Phosphatase; Adrenal Glands; Alanine Transaminase; Alkaline Phosphatase; Animals; Ascorbic Acid Deficiency; Aspartate Aminotransferases; Enzyme Activation; Kidney; Lipid Metabolism; Liver; Male; Rats; Rats, Inbred Strains; Vanadates; Water Pollutants; Water Pollutants, Chemical

Previously we have observed increased specific activities of several lysosomal hydrolases in scorbutic guinea pigs and thus the specificity of this effect was examined in guinea pigs marginally deficient in ascorbic acid (AA). Guinea pigs were fed an AA-deficient diet for 2 weeks to deplete body AA pools and then fed a stock diet containing 0.5 mg AA/g diet or the deficient diet plus oral administration of 10 mg AA/day, 1 mg AA/100 g body weight or 0.5 mg AA/100 g body weight each day. Animal were periodically killed during the 12-week experiment and lysosomes isolated from individual livers and analyzed. Serum and brain AA declined when AA was withheld, returned to normal when the stock diet or 10 mg AA were fed but remained at low levels on administation of 1.0 mg or 0.5 mg AA/100 g body weight. Brain norepinephrine followed a similar pattern to brain AA and was opposite to the pattern observed for dopamine. In guinea pigs receiving 1 mg AA/100 g body weight, amine concentrations slowly returned to normal after 8 weeks. Serum hexosaminidase and lysosomal cathepsins A and B were unchanged during the experiment, whereas lysosomal hexosaminidase and acid phosphatase were significantly higher when the experiment was terminated.

Topics: Acid Phosphatase; Animals; Ascorbic Acid; Ascorbic Acid Deficiency; beta-N-Acetylhexosaminidases; Carboxypeptidases; Cathepsin A; Cathepsin B; Cathepsins; Dose-Response Relationship, Drug; Guinea Pigs; Hexosaminidases; Liver; Lysosomes; Male

The effects of L-ascorbic acid deficiency on guinea pig hepatic and brain lysosomal hydrolases were examined. In general, hepatic beta-N-acetylhexosaminidase, beta-D-glucoronidase, alpha-D-galactosidase, alpha-D-mannosidase, and acid phosphatase were elevated in scorbutic animals. This appears to be independent of the starved state. Brain beta-D-glucoronidase and acid phosphatase followed a similar pattern to that observed with the liver enzymes, but brain beta-N-acetylhexosaminidase was not affected by L-ascorbic acid decreased the activity of hepatic beta-N-acetylhexosaminiadase was unaffected by dietary treatments although the activity of beta-N-acetylhexosaminidase A tended to increase in the scorbutic animals. Subcellular fractions were obtained from the three groups of animals and the recoveries of protein, beta-N-acetylhexosaminidase, and glucose-6-phosphatase estimated.

Topics: Acid Phosphatase; Animals; Ascorbic Acid; Ascorbic Acid Deficiency; Brain; Glycoside Hydrolases; Guinea Pigs; Hydrolases; Liver; Lysosomes; Male; Subcellular Fractions

Enzymes related to bactericidal activities of leukocytes were studied in ascorbic acid deficient guinea pig leukocytes. The activities of glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase were not affected either under resting or phagocytizing conditions in ascorbic acid deficiency. Granule bound NADPH-oxidase activity of resting leukocytes also was not altered in ascorbic acid deficiency. However, the extent of stimulation in NADPH-oxidase activity under phagocytizing condition was found to be significantly lower in ascorbic acid deficient leukocytes than that in control leukocytes. Similary, the extent of release of acid phosphatase from lysosomes during phagocytosis was also low in ascorbic acid deficient leukocytes. Ascorbic acid deficiency did not influence the activities of glutathione reductase and myeloperoxidase of leukocytes. The significance of these enzyme changes is discussed in relation to the decreased phagocytic and bactericidal activities of leukocytes in ascorbic acid deficiency.

Topics: Acid Phosphatase; Animals; Ascorbic Acid Deficiency; Blood Bactericidal Activity; Escherichia coli; Glucosephosphate Dehydrogenase; Glutathione Reductase; Guinea Pigs; NADH, NADPH Oxidoreductases; Neutrophils; Peroxidase; Phagocytosis; Phosphogluconate Dehydrogenase

Topics: Acid Phosphatase; Adrenal Glands; Alkaline Phosphatase; Animals; Ascorbic Acid Deficiency; Aspartate Aminotransferases; Glucose-6-Phosphatase; Guinea Pigs; Hyaluronoglucosaminidase; Immunoelectrophoresis; Kidney; Liver; Male

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Ascorbic Acid; Ascorbic Acid Deficiency; Guinea Pigs; Histocytochemistry; Lipids; Male; Scurvy; Spermatozoa; Succinate Dehydrogenase; Testis

Topics: 5'-Nucleotidase; Acid Phosphatase; Adenosine Monophosphate; Ascorbic Acid Deficiency; Biotin; Folic Acid; Nucleotidases; Protein Tyrosine Phosphatases