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Journal of Chinese Integrative Medicine: Volume 10, 2012   Issue 3,  Pages: 337-346

DOI: 10.3736/jcim20120314
Original Experimental Research
Potential of the homeopathic remedy, Arnica Montana 30C, to reduce DNA damage in Escherichia coli exposed to ultraviolet irradiation through up-regulation of nucleotide excision repair genes
1. Sreemanti Das (Cytogenetics and Molecular Biology Laboratory, Department of Zoology, University of Kalyani, Kalyani 741235, India )
2. Santu Kumar Saha (Cytogenetics and Molecular Biology Laboratory, Department of Zoology, University of Kalyani, Kalyani 741235, India )
3. Arnab De (Cytogenetics and Molecular Biology Laboratory, Department of Zoology, University of Kalyani, Kalyani 741235, India )
4. Durba Das (Cytogenetics and Molecular Biology Laboratory, Department of Zoology, University of Kalyani, Kalyani 741235, India )
5. Anisur Rahman Khuda-Bukhsh (Cytogenetics and Molecular Biology Laboratory, Department of Zoology, University of Kalyani, Kalyani 741235, India E-mail: khudabukhsh_48@rediffmail.com)
OBJECTIVE: To examine to what degree an ultra-highly diluted homeopathic remedy, Arnica Montana 30C (AM-30C), used in the treatment of shock and injury, can modulate the expression of nucleotide excision repair genes in Escherichia coli exposed to ultraviolet (UV) irradiation.
METHODS: E. coli were cultured to their log phase in a standard Luria-Bertani medium and then exposed to sublethal doses of UV irradiation at 25 and 50 J/m2 for 22.5 and 45 s, respectively. The UV-exposed bacteria were then supplemented with either AM-30C (drug) or placebo (P-30C). The drug-treated and placebo-treated bacteria were subjected to assay for DNA damage and oxidative stress 90 min after UV exposure. Several protocols like comet assay, gel electrophoresis for DNA ladder and intracellular reactive oxygen species (ROS) generation, and biomarker measurement like superoxide dismutase (SOD), catalase (CAT) and reduced glutathione (GSH) were conducted. The mRNA expressions of the excision repair genes like ultraviolet repair uvrA, B and C genes (or also known as excision repair genes) were estimated by reverse transcription-polymerase chain reaction method.
RESULTS: The UV-exposed bacteria showed DNA damage and oxidative stress, as revealed by an increase in ROS generation, and a decrease in SOD, CAT and GSH activities. As compared to placebo, the AM-30C-treated bacteria showed less DNA damage and oxidative stress as manifested by a decrease in ROS generation, and an increase in SOD, CAT and GSH activities. AM-30C also up-regulated the expression of repair genes as compared to the control.
CONCLUSION: AM-30C helped repair the DNA damage through up-regulation of repair genes and also ameliorated the oxidative stress through the reduction of ROS generation and suitable modulation of anti-oxidative stress enzymes.
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Das S, Saha SK, De A, Das D, Khuda-Bukhsh AR. Potential of the homeopathic remedy, Arnica Montana 30C, to reduce DNA damage in Escherichia coli exposed to ultraviolet irradiation through up-regulation of nucleotide excision repair genes. J Chin Integr Med / Zhong Xi Yi Jie He Xue Bao. 2012; 10(3): 337-346.
1Lubin D, Jensen EH. Effects of clouds and stratospheric ozone depletion on ultraviolet radiation trends[J].Nature, 1995, 377(6551): 710-713.  
2Taylor JS. DNA, sunlight, and skin cancer[J].Pure Appl Chem, 1990, 67(1): 183-190.  
3Wiseman H, Halliwell B. Damage to DNA by reactive oxygen and nitrogen species: role in inflammatory disease and progression to cancer[J].Biochem J, 1996, 313(Pt 1): 17-29.  
4Valko M, Rhodes CJ, Moncol J, Izakovic M, Mazur M. Free radicals, metals and antioxidants in oxidative stress-induced cancer[J].Chem Biol Interact, 2006, 160(1): 1-40.  
5Halliwell B, Gutteridge J. Free radicals in biology and medicine. 4th ed. Oxford: Oxford University Press. 2007.
6Sinha RP, Hder DP. UV-induced DNA damage and repair: a review[J].Photochem Photobiol Sci, 2002, 1(4): 225-236.  
7Solomon KR. Effects of ozone depletion and UV-B radiation on humans and the environment[J].Atmos Ocean, 2008, 46(1): 185-202.  
8Cooper GM. The cell: a molecular approach. Washington: Sinauer Associates Inc. 1997.
9Ravanat JL, Douki T, Cadet J. Direct and indirect effects of UV radiation on DNA and its components[J].J Photochem Photobiol B, 2001, 63(1-3): 88-102.  
10de Lima-Bessa KM, Armelini MG, Chiganas V, Jacysyn JF, Amarante-Mendes GP, Sarasin A, Menck CF. CPDs and 6-4PPs play different roles in UV-induced cell death in normal and NER-deficient human cells[J].DNA Repair (Amst), 2008, 7(2): 303-312.  
11E. coli K-12[J].Proc Natl Acad Sci U S A, 1964, 51: 293-300.  
12Lin JJ, Sancar A. Reconstitution of nucleotide excision nuclease with UvrA and UvrB proteins from Escherichia coli and UvrC protein from Bacillus subtilis. J Biol Chem. 1990; 265(34): 21337-21341.
13Escherichia coli. Disassembly of the UvrBC-DNA complex by helicase Ⅱ and DNA polymerise Ⅰ[J].J Biol Chem, 1992, 267(2): 780-788.  
14Pettijohn DE, Hanawalt P. Evidence for repair replication of ultraviolet damaged DNA in bacteria[J].J Mol Biol, 1964, 9: 395-410.  
15Boericke W. Pocket manual of homeopathic materia medica. Calcutta: Sett Dey and Co. 1976.
16Mitra K, Kundu SN, Khuda-Bukhsh AR. Efficacy of a potentized homoeopathic drug (Arsenicum Album-30) in reducing toxic effects produced by arsenic trioxide in mice: Ⅰ. On rate of accumulation of arsenic in certain vital organs[J].Complement Ther Med, 1998, 6(4): 178-184.  
17Khuda-Bukhsh AR. Potentized homoeopathic drugs act through regulation of gene-expression: a hypothesis to explain their mechanism and pathways of action in vitro[J].Complement Ther Med, 1997, 5(1): 43-46.  
18Biswas SJ, Khuda-Bukhsh AR. Effect of a homeopathic drug, Chelidonium, in amelioration of p-DAB induced hepatocarcinogenesis in mice[J].BMC Complement Altern Med, 2002, 2: 4.  
19Chakrabarti J, Biswas SJ, Khuda-Bukhsh AR. Cytogenetical effects of sonication in mice and their modulations by actinomycin D and a homeopathic drug, Arnica 30[J].Indian J Exp Biol, 2001, 39(12): 1233-1242.  
20Khuda-Bukhsh AR. Laboratory research in homeopathy: pro[J].Intregr Cancer Ther, 2006, 5(4): 320-332.  
21Khuda-Bukhsh AR, Pathak S. Homeopathic drug discovery: theory update and methodological aspect[J].Expert Opin Drug Discov, 2008, 3(8): 979-990.  
22Chikramane PS, Suresh AK, Bellare JR, Kane SG. Extreme homeopathic dilutions retain starting materials: A nanoparticulate perspective[J].Homeopathy, 2010, 99(4): 231-242.  
23Saha S, Jana B, Basu T. The two inducible responses, SOS and heat-shock, in Escherichia coli act synergistically during Weigle reactivation of the bacteriophage phiX174. Int J Radiat Biol. 2007; 83(7): 463-469.
24Kakkar P, Das B, Viswanathan PN. A modified spectrophotometric assay of superoxide dismutase[J].Indian J Biochem Biophys, 1984, 21(2): 130-132.  
25Omran RG. Peroxide levels and the activities of catalase, peroxidase, and indoleacetic acid oxidase during and after chilling cucumber seedlings[J].Plant Physiol, 1980, 65(2): 407-408.  
26Sedlak J, Lindsay RH. Estimation of total, protein-bound, and nonprotein sulfhydryl groups in tissue with Ellman’s reagent[J].Anal Biochem, 1968, 25(1): 192-205.  
27Dhawan A, Mathur N, Seth PK. The effect of smoking and eating habits on DNA damage in Indian population as measured in the Comet assay[J].Mutat Res, 2001, 474(1-2): 121-128.  
28Chowdhury R, Dutta A, Chaudhuri SR, Sharma N, Giri AK, Chaudhuri K. In vitro and in vivo reduction of sodium arsenite induced toxicity by aqueous garlic extract[J].Food Chem Toxicol, 2008, 46(2): 740-751.  
29Dhiman R, Kathania M, Raje M, Majumdar S. Inhibition of bfl-1/A1 by siRNA inhibits mycobacterial growth in THP-1 cells by enhancing phagosomal acidification[J].Biochim Biophys Acta, 2008, 1780(4): 733-742.  
30Sies H, Menck CF. Singlet oxygen induced DNA damage[J].Mutat Res, 1992, 275(3-6): 367-375.  
31Sies H. Damage to plasmid DNA by singlet oxygen and its protection[J].Mutat Res, 1993, 299(3-4): 183-191.  
32Yao XH, Min H, Lv ZM. Response of superoxide dismutase, catalase, and ATPase activity in bacteria exposed to acetamiprid[J].Biomed Environ Sci, 2006, 19(4): 309-314.  
33Imlay JA, Chin SM, Linn S. Toxic DNA damage by hydrogen peroxide through the Fenton reaction in vivo and in vitro[J].Science, 1988, 240(4852): 640-642.  
34Chen JR, Weng CN, Ho TY, Cheng IC, Lai SS. Identification of the copper-zinc superoxide dismutase activity in Mycoplasma hyopneumoniae[J].Vet Microbiol, 2000, 73(4): 301-310.  
35Gerlach D, Reichardt W, Vettermann S. Extracellular superoxide dismutase from Streptococcus pyogenes type 12 strain is manganese-dependent[J].FEMS Microbiol Lett, 1998, 160(2): 217-224.  
36Jung S. Expression level of specific isozymes of maize catalase mutants influences other antioxidants on norflurazon-induced oxidative stress[J].Pestic Biochem Physiol, 2003, 75(1-2): 9-17.  
37Khuda-Bukhsh AR. Towards understanding molecular mechanisms of action of homeopathic drugs: an overview[J].Mol Cell Biochem, 2003, 253(1-2): 339-345.  
38Khuda-Bukhsh AR. Mice as a model for homeopathy research[J].Homeopathy, 2009, 98(4): 267-279.  
39Mallick P, Mallick JC, Guha B, Khuda-Bukhsh AR. Ameliorating effect of microdoses of a potentized homeopathic drug, Arsenicum Album, on arsenic-induced toxicity in mice[J].BMC Complement Altern Med, 2003, 3: 7.  
40Bhattacharyya SS, Mandal SK, Biswas R, Paul S, Pathak S, Boujedaini N, Belon P, Khuda-Bukhsh AR. In vitro studies demonstrate anticancer activity of an alkaloid of the plant Gelsemium sempervirens[J].Exp Biol Med (Maywood), 2008, 233(12): 1591-1601.  
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