RADIOPROTECTIVE ACTIVITY OF FICUS RACEMOSA ETHANOL EXTRACT AGAINST ELECTRON BEAM INDUCED DNA DAMAGE IN VITRO, IN VIVO AND IN SILICO
Keywords:Ficus racemosa, Radioprotective activity, Molecular docking, p53 protein
Objective: To investigate the radioprotective effect of Ficus racemosa (Fr) ethanol stem bark extract against electron beam radiation (EBR) induced DNA damage using in vitro, in vivo and in silico models.
Methods: The extract of Fr was tested against radiation induced DNA damage by exposing pBR322 plasmid to different EBR dose rates. Comet assay was conducted using mice which were exposed at 6Gy EBR. In silico study was performed by inhibiting p53 protein C-chain (1TUP C) using phyto chemicals of Fr.
Results: The in vitro results revealed that, Fr at lower concentration (50Âµg) showed inhibitory effect on radiation induced DNA damage compared with control. Exposure of mice to 6Gy EBR increased comet parameters like TL (Tail length), OTM (Olive tail moment) and %T (percentage of DNA in the tail) of blood lymphocytes. Fr ethanol extract given orally prior to irradiation at a dose of 400 mg/kg body weight protected the DNA from the radiation damage. The phytochemicals of Fr showed clear interaction with p53 protein chain C, specifically binding to Arginine 248 (ARG248) and Arginine 273 (ARG273) amino acid residues thereby inhibiting the p53 protein-DNA interaction upon radiation.
Conclusion: The present study indicates that Fr ethanol extract significantly reduced radiation induced DNA damage in vivo and in vitro. It also showed that the biologically active compounds of Fr have ability to inhibit wild p53 protein which is responsible for apoptosis; these compounds can be used as radioprotectors during chemotherapy to protect normal tissues surrounding cancerous tissue.
Puthran SS, Sudha K, Rao GM, Shetty BV. Oxidative stress and low dose ionizing radiation. Indian J Physiol Pharmacol 2009;53(2):181â€“4.
Thulasi GP, Veena PS, Dharmendra KM, Cherupally KN, Janardhanan KK. Prevention of radiation-induced damages by aqueous extract of Ganoderma lucidum occurring in southern parts of India. Curr Sci 2006;91:3.
Qimin Z, Tsuen C, Michael JA, Albert JF. Tumor suppressor p53 can participate in transcriptional induction of the GADD45 promoter in the absence of direct DNA binding. Mol Cell Biol 1998;18(5):2768â€“78.
Amir E, Haim R, Yael DP, Remo R, Zippora S. Structural studies of p53 inactivation by DNA-contact mutations and its rescue by suppressor mutations via alternative protein DNA interactions. Nucleic Acids Res 2013;41:1â€“12.
Prasanna PG, Stone HB, Wong RS, Capala J, Bernhard EJ, Vikram B, et al. Normal tissue protection for improving radiotherapy. Transl Lung Cancer Res 2012;1:35-48.
Arora R, Chawla R, Singh S, Kumar R, Sharma AK, Puri SC, et al. Radioprotection by himalayan high altitude region plants. In: herbal drugs: A Twenty-first century perspective. Jaypee Brothers Medical Publishing Limited, New Delhi; 2006. p. 301-25.
Arora R, Chawla R, Singh S, Sagar RK, Kumar R, Sharma A, et al. Bio-prospection for radioprotective molecules from indigenous flora. Phytomed 2006;16:179-219.
Satish AB, Deepa RV, Nikhil CT, Vinodkumar SD, Saurabh ST. Ficus racemosa Linn: A comprehensive review. J Appl Chem 2014;3(4):1423-31.
Murti K, Kumar U, Panchal M, Shah M. Exploration of preliminary phytochemicals studies of roots of Ficus racemosa. Marmara Pharm J 2011;15:80-3.
Miller LC, Tainter MC. Estimation of LD50 and its errors by means of logarithmic probity graph paper. Proc Soc Exp Biol Med 1944;57:261-4.
Solanki ND, Bhavsar SK. Toxicological evaluation of aqueous fraction of Ficus racemosa and bauhinia variegate bark in rats. Int J Toxicol Pharmacol Res 2014;6(4):80-5.
Retimen S, Frankel SA. Colorimetric method for determination of serum glutamic oxaloacetic and glutamic pyruvate transaminases. Am J Clin Pathol 1957;28:56-63.
King EJ, Armstrong AR. Determination of serum and bile phosphatase activity. Canadian Med Assoc J 1934;31:56-63.
Malloy HT, Evelyn KA. The determination of bilirubin with the photoelectric colorimeter. J Biol Chem 1937;119:481-90.
Madhu LN, Suchetha KN, Vijay R. Validation of DNA damage progression with days after single exposure of sub-lethal dosage of electron beam radiation. J Appl Pharm Sci 2012;2(12):23-6.
Singh NP. Microgel for estimation of DNA strand breaks DNA protein crosslinkâ€™s and apoptosis. Mutat Res 2000;455:111â€“27.
Rogers D, Hopfinger AJ. Application of genetic function approximation to quantitative structure activity relationships and quantitative structure property relationships. J Chem Inf Comput Sci 1994;34:854.
Gade DR, Pavan KKNV, Duganath N, Raavi D, Kancharla A. ADMET Docking studies and binding energy calculations of some novel ACE inhibitors for the treatment of diabetic nephropathy. Int J Drug Dev Res 2012;4(3):268-82.
Daisy P, Suveena S. Solutions to pharmaceutical issues for anti-cancer drugs by Accord excel. Asian J Pharm Clin Res 2012;5:149-58.
Veerapur VP, Prabhakar KR, Parihar VK, Kandadi MR, Ramakrishna S, Mishra B, et al. Ficus racemosa stem bark extract: A potent antioxidant and a probable natural radioprotector. J Evidence-Based Complementary Altern Med 2009;6:317-24.
Kramer B, Rarey M, Lengauer T. Evaluation of the FLEXX: Incremental construction algorithm for protein ligand docking. Proteins 1999;37(2):228-41.
Maurya DK, Salvi VP, Nair CKK. Radioprotection of normal tissues in tumor bearing mice by Troxerutin. J Radiat Res 2004;45:221-8.
Patwardhan B, Vaidya ADB, Chorghade M. Ayurveda and natural products drug discovery. Curr Sci 2004;86:789-99.
Nitin MG, Cherupally KN. Radiation protection by diethyldithicarbamate: Protection of membrane and DNA in vitro and in vivo against Î³-radiation. J Radiat Res 2004;45(2):175-80.
Cooke MS, Evans MD, Dizdaroglu M, Lunec J. Oxidative DNA damage: Mechanism mutation and disease. FASEB J 2003;17:1195â€“214.
Sarfaraz A, Feroz K. QSAR and docking studies on xanthone derivatives for anticancer activity targeting DNA topoisomerase IIÎ±. Drug Design Dev Ther 2014;8:183â€“95.
Jayasimha RD, Muralidhara RD, Rao DS. Phytochemicals screening and in silico approach for the identification of anti-stress compounds from medicinal plants. Int J Appl Biol Pharm Technol 2013;4:324-34.
Venkataramana CHS, Ramya KMS, Swetha SS, Madhavan V. In silico ADME and toxicity studies of some novel indole derivatives. J Appl Pharm Sci 2011;1:159-62.
Durga DM, Dibyabhaba P, Manne M, Amineni U. Implementation of computational methods for designing potential inhibitors against human p38Î± protein. Nature proceedings; 2010.
Dixon SL, Merz KM. One dimensional molecular representation and similarity calculations: Methodology and validation. J Med Chem 2001;44:3795-809.
Lin X, Ramamurthy K, Mishima M, Kondo A, Howell SB. p53 interacts with the DNA mismatch repair system to module the cytotoxicity and mutagenicity of hydrogen peroxide. Mol Pharmacol 2000;58:1222-9.
Geisler S, Borresen DAL, Johnson H, Aas T, Geisler J, Akslen LA, et al. TP53 gene mutations predict the response to neoadjuvant treatment with 5-fluorouracil and mitomycin in locally advanced breast cancer. Clin Cancer Res 2003;9:5582-8.
Bykov VJN, Issaeva N, Shilov A, Hultcrantz M, Pugacheva E, Chumakov P, et al. Restoration of the tumor suppressor function to mutant p53 by a low molecular weight compound. Nat Med 2002;2:282-8.