Abstract
The evolving role of mitochondria, in mediating chemotherapy-induced apoptosis motivated us for the studies described here. The combination of cisplatin and cyclophosphamide is widely used in treating various types of cancers. The purpose of our study was to understand the mechanism of the toxicity induced by the co-administration of cisplatin and cyclophosphamide, on mitochondrial bioenergetics, and to study the protective effect of prior administration of the medicinal plant extract Phyllanthus fraternus. Our results reveal that co-administration of cisplatin (12 mg/kg, i.p) and cyclophosphamide (150 mg/kg, oral) to wistar rats (100 g) significantly alters mitochondrial structure and hence function. The rate of mitochondrial respiration was decreased significantly with both NAD + and FAD-linked substrates. The respiratory control ratio, an index of membrane integrity and the P/O ratio, a measure of phosphorylating efficiency also were decreased significantly accompanied by elevation in the lipid peroxide levels in liver, kidney homogenate and liver mitochondria respectively. Also, the phospholipid content of the mitochondrial membrane, showed a significant decrease, indicating mitochondrial membrane changes. Prior administration of an aqueous extract of P. fraternus (100 mg/kg) to rats, showed protection on all parameters investigated. Administration of P. fraternus alone did not show any significant changes on mitochondrial membrane bioenergetics. Thus, we propose, that the toxic side effects of cisplatin + cyclophosphamide, are due to a chain of interconnected events, within the mitochondrial inner membrane, ultimately leading to hepatotoxicity and nephrotoxicity. Further, our work also suggests that administration of aqueous extract of P. fraternus can enhance the therapeutic potential of anticancer drugs by reducing drug related toxicity.
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Ali BH, Moundhri MS (2006) Agents ameliorating or augmenting the nephrotoxicity of cisplatin and other platinum compounds: a review of some recent research. Food Chem Toxicol 44:1173–1183, and their weapons. J Cell Physiol 2002;192:131–7
Anders MW, Robotham JL, Sheu SS (2006) Mitochondria: new drug targets for oxidative stress-induced diseases. Expert Opin Drug Metab Toxicol 2(1):71–79
Bhattacharjee R, Sil PC (2006) The protein fraction of Phyllanthus niruri plays a protective role against acetaminophen induced hepatic disorder via its antioxidant properties. Phytother Res 20(7):595–601
Bligh EG, Dyer WJ (1959) A rapid method of total lipid extraction and purification. Can J Biochem physiol 37:911–917
Calixto JB, Santos AR, Cechinel FV, Yunes RA (1998) Review of the Plants of the Genus Phyllanthus: Their Chemistry, Pharmacology, and Therapeutic Potential. Med Res Rev 8(4):225–258
Chatterjee M, Sil PC (2006) Hepatoprotective effect of aqueous extract of Phyllanthus niruri on nimesulide-induced oxidative stress in vivo. Indian J Biochem Biophys 43(5):299–305
Cooperstein ST, Liazarow AJ (1951) A microspectrophotometric method for the determination of cytochrome c oxidase. J Biol Chem 189:665–670
Custódio JB, Cardoso CM, Santos MS, Almeida LM, Vicente JA, Fernandes MA (2009) Cisplatin impairs rat liver mitochondrial functions by inducing changes on membrane ion permeability: prevention by thiol group protecting agents. Toxicology 2,259(1–2):18–24
Davies MJ, Fu S, Dean RT (1995) Protein hydroperoxides can give rise to reactive free radicals. Biochem 305:643–649
Estabrook RW (1967) Mitochondrial respiratory control and the polarographic measurements of ADP:O ratios. In: Estabrook RW, Pullman ME (eds) Methods in enzymology, 10th edn. Academic, New York, pp 41–47
Fiske CH, Subba Row Y (1925) The colorimetric determination of phosphorus. J Biol Chem 66:375–400
Gemba M, Fukuishi N, Nakano S (1988) Effect of N-N-diphenyl-p-phenylenediamine pretreatment on urinary enzyme exctretion in cisplatin nephrotoxicity in rats. Jpn J Pharmacol 46:90–92
Gornall AG, Bardawill CJ, David MM (1949) Determination of serum proteins by means of the biuretic reaction. J Biol Chem 177:751–766
Hanaki Y, Sugiyama S, Akiyamam N, OzawaT (1990) Role of the autonomic nervous system in cyclophosphamide induced heart mitochondrial dysfunction in rats. Biochem Int 21: 289–295
Hanigan MH, Devarajan P (2003) Cisplatin nephrotoxicity: molecular mechanisms. Cancer Ther 1:47–61
Hatefi Y, Reiske JS (1967) In: Estabrook RE, Pullman ME (eds) Methods in enzymology, 10. Academic, New York, pp 225–231
Kameyama Y, Gemba M (1991) The iron chelator deferoxamine prevents cisplatin-induced lipid peroxidation in rat kidney cortical slices. Jpn J Pharmacol 57: 259–262
Kearney EB (1957) Studies on succinate dehydrogenase IV ACTIVATION OF THE BEEF HEART ENZYME. J Biol Chem 229:363–375
King TE (1967) Preparation of succinate dehydrogenase and reconstitution of succinate oxidase. In: Estabrook RW, Pullman ME (eds) Methods in enzymology, 10. Academic press Inc, New York, pp 322–331
King TE, Robert HL (1967) Preparation and properties of soluble NADH dehydrogenase from cardiac muscle. In: Estabrook RW, Pullman ME (eds) Methods in Enzymology, 10. Academic, Newyork, pp 275–294
Kozel’tsev VL, Golubev VP, Volodina TV (1980) Protective role of phospholipids in the enzymatic degradation of mitochondrial membrane proteins. Vopr Med Khim 26(5):680–5
Lawrence CB, Davis NT (1986) A novel simple and rapid method for the isolation of mitochondria which exibit respiratory control from small intestinal mucosa. Biochem Biophys Acta 848:35–40
Levrat C, Louisot P (1994) Study of the succinate dehydrogenase activation in permeabilized mitochondria through the Ca(2+)-stimulated phospholipase A2. Biochem Mol Biol Int 34(3):569–578
Manjrekar AP, Jisha V, Bag PP, Adhikary B, Pai MM, Hegde A, Nandini M (2008) Effect of Phyllanthus niruri Linn. treatment on liver, kidney and testes in CCl4 induced hepatotoxic rats. Indian J Exp Biol 46(7):514–20
Martins NM, Santos NAG, Curti C, Bianchi MLP, Santos AC (2008) Cisplatin induces mitochondrial oxidative stress with resultant energetic metabolism impairment, membrane rigidification and apoptosis in rat liver. J Appl Toxicol 28:337–344
Ogawa Y, Kondo T, Sugiyama S, Ogawa K, Satake T, Ozawa T (1987) Role of phospholipase in the genesis of doxorubicin-induced cardiomyopathy in rats. Cancer Res 47:1239–1243
Ohkawa H, Ohishi N, Yagi K (1979) Assay for lipid peroxides in animal tissue by thiobarbituric acid reaction. Annal Biochem 95:351–358, 67
Padma P, Setty OH (1997) Protective effect of vitamin E against ethionine toxicity. Biochem Mol Biol Int 41:785–795
Pinmai K, Chunlaratthanabhorn S, Ngamkitidechakul C, Soonthornchareon N, Hahnvajanawong C (2008) Synergistic growth inhibitory effects of Phyllanthus emblica and Terminalia bellerica extracts with conventional cytotoxic agents: doxorubicin and cisplatin against human hepatocellular carcinoma and lung cancer cells. World J Gastroenterol 14:1491–1497
Pompella A, Romani A, Benedetti A, Comporti M (1991) Loss of membrane protein thiols and lipid peroxidation in allyl alcohol hepatotoxicity. Biochem Pharmacol 41:1255–1259
Pozzan T, Miconi V, Di Virgilio F, Azzone GF (1979) H+/site, charge/site, and ATP/site ratios at coupling sites I and II in mitochondrial e- transport. J Biol Chem 254:12000-12005
Sailaja R, Setty OH (2006) Protective effect of Phyllanthus fraternus against allyl alcohol-induced oxidative stress in liver mitochondria. J Ethnopharmacol 21:105(1–2):201–9
Sangeetha P, Das UN, Koratkar R, Suryaprabha P (1990) Increase in free radical generation and lipid peroxidation following chemotherapy in patients with cancer. Free Radic Biol Med 8: 15–19
Santiago E, López-Moratalla N, Segovia JL, Eugui J (1977) NADH-cytochrome c reductase, succinate cytochrome c reductase and phospholipids. Rev Esp Fisiol 33(3):197–204
Santos NAG, Martins NM, Curti C, Bianchi MLP, Santos AC (2007) Dimethylthiourea protects against mitochondrial oxidative damage induced by cisplatin in liver of rats. Chem Biol Int 170:177–186
Sebastian T, Setty OH (1999) Protective effect of P.fraternus against ethanol induced mitochondrial function. Alcohol 17:29–34
Shanafelt TD, Lin T, Geyer SM et al (2007) Pentostatin, cyclophosphamide, and rituximab regimen in older patients with chronic lymphocytic leukemia. Cancer 109(11):2291–2298
Simpson JA, Narita S, Gieseg S, Gebicki S, Gebicki JM, Dean RT (1992) Long-lived reactive species on free-radical-damaged proteins. Biochem J 282:621–624
Somani SM, Husain K, Whitworth C, Trammell GL, Malafa M, Rybak LP (2000) Dose-dependent protection by lipoic acid against cisplatin induced nephrotoxicity in rats: antioxidant defense system. Pharmacol Toxicol 86:234–241
Stankiewicz A, Skrzydlewska E, Makieła M (2002) Effects of amifostine on liver oxidative stress caused by cyclophosphamide administration to rats. Drug Metabol Drug Interact 19:67–82
Sugihara KS, Nakano S, Koda M, Tanaka K, Fukuishi N, and Gemba M (1987) Stimulatory effect of cisplatin on production of lipid peroxidation in renal fissues. Jpn J Pharmacol 43: 247–252
Sugiyama S, Hayakawa M, Kato T, Hanaki Y, Shimizu K, Ozawa T (1989) Adverse effects of anti-tumor drug, cisplatin, on rat kidney mitochondria: disturbances in glutathione peroxidase activity. Biochem Biophys Res Commun 159:1121–1127
Supratim Ray, Chandana Sengupta, Kumal Roy (2005) Evaluation of Ascorbic acid as suppressor of cyclophospamide induced lipid peroxidation using common laboratory markers. Acta Poloniae Pharmaceutican Drug Res 62(2):145–151
Szewczyk A, Wojtczak L (2002) Mitochondria as a pharmacological target. Pharmacol Rev 54:101–127
Tisdale HD (1967) In: Estabrook RW, Pullman ME (eds) Methods in enzymology, 10. Academic, New York, pp 213–215
Tzagoloff, A., and Myers, A. M. (1986) Genetics of mitochondrial biogenesis. Annu Rev Biochem 55: 249–285
Yonetani T (1967) Methods in Enzymology. In: Estrabrook RW, Pullman ME (eds) Cytochrome oxidase: beef heart. 10. Academic Press, New york, pp 332–335
Young SD, Whissell M, Noble JC et al (2006) Phase II clinical trial results involving treatment with low-dose daily oral cyclophosphamide, weekly vinblastine, and rofecoxib in patients with advanced solid tumors. Clin Cancer Res 12(10):3092–3098
Yu L, Yu CA, King TE (1978) The indispensability of phospholipid and ubiquinone in mitochondrial electron transfer from succinate to cytochrome c. J Biol Chem 253:2657–2663
Zhang JG, Lindup WE (1994) Cisplatin nephrotoxicity: decreases in mitochondrial protein sulphydryl concentration and calcium uptake by mitochondria from rat renal cortical slices. Biochem Pharmacol 47:1127–1135
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Kumari, K.K., Setty, O.H. Protective effect of Phyllanthus fraternus against mitochondrial dysfunction induced by co-administration of cisplatin and cyclophosphamide. J Bioenerg Biomembr 44, 179–188 (2012). https://doi.org/10.1007/s10863-012-9423-6
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DOI: https://doi.org/10.1007/s10863-012-9423-6