Abstract
The present investigation was undertaken to study the effect of diclofenac sodium on alkaline phosphatase and acid phosphatase activity in mice tibiae. Diclofenac was administered at the dose rate of 10 mg/kg body wt/day intramuscularly to experimental animals for 10, 20 and 30 days while the control group received normal saline. Alkaline phosphatase and acid phosphatase enzymes were assayed biochemically. Acid phosphatase activity was studied by histochemical analysis whereas alkaline phosphatase was analyzed by native PAGE. Student’s t test and 2-way Anova were applied wherever required. Significant decrease in acid phosphatase was revealed during this course of study which was recorded as 23.03, 22.06 and 15.72 % after 10, 20 and 30 days respectively. Similar trend of decreased acid phosphatase was demonstrated during histochemical study along with remarkable histopathological changes after diclofenac treatment for various durations of investigation. A down-regulation of 28.94 % was recorded in alkaline phosphatase activity of tibia after 10 days whereas no significant change was observed after 20 days. Slight increase (5.12 %) in alkaline phosphatase activity was documented after 30 days of drug therapy. Results from biochemical analysis of alkaline phosphatase were further supported by electrophoretic studies.
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Green RE, Newton I, Shultz S, Cunningham AA, Gilbert M, Pain DJ, Prakash V (2004) Diclofenac poisoning as a cause of vulture population declines across the Indian subcontinent. J Appl Ecol 41:793–800
McComarck K, Brune K (1991) Dissociation between the antinociceptive and anti-inflammatory effects of the nonsteroidal anti-inflammatory drugs. A survey of their analgesic efficacy. Drugs 41:533–547
Nair P, Kanwar SS, Sanyal SN (2006) Effects of non steroidal anti-inflammatory drugs on the antioxidant defense system and the membrane functions in the rat intestine. Nutr Hosp 21:638–649
Mizuno H, Liang RF, Kawabata A (1990) Effects of oral administration of various anti-inflammatory drugs on bone growth and bone wound healing in mice. Meikai Daigaku Shigaku Zasshi 19:234–250
Krischak GD, Augat P, Blakytny R, Claes L, Kinzl L, Beck A (2007) The non-steroidal anti-inflammatory drug diclofenac reduces appearance of osteoblasts in bone defect healing in rats. Arch Orthop Trauma Surg 127:453–458
Harris H (1989) The human alkaline phosphatase: what we know and what we don’t know. Clin Chim Acta 186:133–150
Safadi A, Livne E, Silbermann M, Reznick AZ (1991) Activity of alkaline phosphatase in rat skeletal muscle localized along the sarcolemma and endothelial cell membranes. J Histochem Cytochem 39:199–203
Halleen JM, Raisanen S, Salo JJ, Reddy SV, Roodman GD, Hentunen TA, Lehenkari PP, Kaija H, Vihko P, Vaananen HK (1999) Intracellular fragmentation of bone resorption products by reactive oxygen species generated by osteoclastic tartrate-resistant acid phosphatase. J Biol Chem 274:22907–22910
Vaaraniemi J, Halleen JM, Kaarlonen K, Ylipahkala H, Alatalo SL, Andersson G, Kaija H, Vihko P, Vaananen HK (2004) Intracellular machinery for matrix degradation in bone-resorbing osteoclasts. J Bone Miner Res 19:1432–1440
Urena P, de Vernejoul MC (1999) Circulating biochemical bone markers of bone remodeling in uremic patients. Kidney Int 55:2141–2156
Mclean I, Nakane P (1974) Periodate-lysine-paraformaldehyde fixative: a new fixative for immunoelectron microscopy. J Histochem Cytochem 22:1077–1083
Mori S, Sawai T, Teshima T, Kyogoku M (1988) A new decalcifying technique for immunohistochemical studies of calcified tissue, especially applicable to cell surface marker demonstration. J Histochem Cytochem 36:111–114
Borque WT, Gross M, Hall BK (1993) A histological technique that preserves the integrity of calcified tissues (bone, enamel), yolky amphibian embryos, and the growth factor antigens in skeletal tissues. J Histochem Cytochem 41:1429–1434
Barka T, Anderson PJ (1962) Histochemical methods for acid phosphatase using hexazonium pararosanilin as coupler. J Histochem Cytochem 10:741–753
Alabi OA, Sanni M (2003) Effect of peroxidized soyabeans oil on acid and alkaline phosphatase activity of some rat bones. Biokemistri 15:76–83
Weil L, Russel MA (1940) Study on plasma phosphatase activity in relation to fat metabolism in rats. J Biol Chem 136:9–22
Tausky HH, Shorr E (1953) A microcolorimetric method for determination of inorganic phosphorous. J Biol Chem 207:675–685
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurements with folin–phenol reagent. J Biol Chem 193:265–275
Fraki J, Ruuskanen O, Kouvalainen J (1977) Biochemical characterization of alkaline phosphatase in guinea pig thymus. Biochim Biophys Acta 482:370–378
Kornblatt MJ, Klugerman A, Nagy F (1983) Biochemical characterization and localization of alkaline phosphatase activity in rat testes. Biol Reprod 29:157–164
Epstein E, Wolf PL, Hortwitz JP, Zak B (1967) An indigogenic reaction for alkaline phosphatase in disc electrophoresis. Am J Clin Pathol 48:530–534
Manchenko GP (2003) Handbook of detection of enzymes on electrophoretic gels. CRC Press, Boca Raton
Boelsterli UA (2002) Mechanisms of NSAID-induced hepatotoxicity: focus on nimesulide. Drug Saf 25:633–648
Feenstra J, Grobbee DE, Mosterd A, Stricker BH (1997) Adverse cardiovascular effects of NSAIDs in patients with congestive heart failure. Drug Saf 17:166–180
Muller SS, Curcelli EC, Sardenberg T, Zuccon A, Junior JLDC, Padovani CR (2004) Clinical and biomechanical analysis of the effect of diclofenac sodium in tibial fracture healing in rats. Acta Ortho Bras 12:197–204
Sassioto MC, Inouye CM, Aydos RD, de Figueiredo AS (2006) Bone repair in rats treated with sodic diclofenac and calcitonin. Acta Cir Bras 21:40–44
Katagiri T, Takahashi N (2002) Regulatory mechanisms of osteoblast and osteoclast differentiation. Oral Dis 8:147–159
Baron R, Neff L, Tran VP, Nefussi J, Vignery A (1986) Kinetic and cytochemical identification of osteoclast precursors and their differentiation into multinucleated osteoclasts. Am J Pathol 2:363–378
Karakawa A, Sano T, Amano H, Yamada S (2010) Inhibitory mechanism of non-steroidal anti-inflammatory drugs on osteoclast differentiation and activation. J Oral Biosci 52:119–124
da Silva RA, Fagundes DJ, Brochado Antoniolli Silva ACM, Sisti KE, de Carvalho TMMB, e Silva DN (2008) Effect of anti-inflammatory agents on the integration of autogenous bone graft and bovine bone devitalized matrix in rats. Acta Cir Bras 23:140–148
Kotake S, Yago T, Kawamoto M, Nanke Y (2010) Effects of nsaids on differentiation and function of human and murine osteoclasts: crucial ‘human osteoclastology’. Pharmaceuticals 3:1394–1410
Karashima M, Fujikawa Y, Itonaga I, Takita C, Tsumura H (2009) The effect of selective cyclooxygenase-2 inhibitor on osteoclast precursors to influence osteoclastogenesis in vitro. Mod Rheumatol 19:192–198
de Araujo MB, Voltarelli FA, Contarteze RVL, Manchado-Gobatto FDB, de Mello MAR (2009) Oxidative stress in rats exercised at different intensities. J Chin Clin Med 4:11–18
Pennington RJT (1964) Disorder of voluntary muscle. Churchill Livingstone, London
Daluiski A, Ramsey KE, Shi Y, Bostrom MP, Nestor BJ, Martin G, Hotchkiss R, Stephen DA (2006) Cyclooxygenase inhibitors in human skeletal fracture healing. Orthopedics 29:259–261
Khoker MA, Dandona P (1988) The effect of indomethacin and aspirin on alkaline phosphatase secretion and [3H] thymidine incorporation by human osteoblasts. Br J Rheumatol 27:291–294
Takiguchi T, Kobayashi M, Nagashima C, Yamaguchi A, Nishihara T, Hasegawa K (1999) Effect of prostaglandin E2 on human recombinant bone morphogenetic protein-2- stimulated osteoblastic differentiation in human periodontal ligament cells. J Periodontal Res 34:431–436
Chouhan S, Sharma S (2011) Subchronic diclofenac sodium induced alterations of alkaline phosphatase in serum and skeletal muscle of mice. Indian J Exp Biol 49:446–454
Ho ML, Cheng JK, Chuang LY, Hsu HK, Wang GJ (1999) Effects of nonsteroidal anti-inflammatory drugs and prostaglandins on osteoblastic functions. Biochem Pharmacol 58:983–990
Sharma S, Malhotra RK (1992) Acid and alkaline phosphatase activities in normal, denervated and exercised chick skeletal muscles. Proc Nat Acad Sci India 62(B)II:155–160
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Chouhan, S., Sharma, S. Down-Regulation of Acid and Alkaline Phosphatases Induced After Prolonged Diclofenac Use in Experimental Mouse Bone. Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci. 84, 37–45 (2014). https://doi.org/10.1007/s40011-013-0191-9
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DOI: https://doi.org/10.1007/s40011-013-0191-9