In-vitro and In-vivo Antioxidant Activity of Ethanol Leaf Extract of Justicia carnea
International Journal of Biochemistry Research & Review,
Page 48-60
DOI:
10.9734/ijbcrr/2020/v29i430185
Abstract
This study investigated the in-vitro antioxidant activity of ethanol leaf extract of Justicia carnea and its effect on antioxidant status of alloxan-induced diabetic albino rats. The in-vitro antioxidant activity was assayed by determining the total phenol, flavonoids, ascorbic acid, β-carotene and lycopene contents and by using 2,2 diphenyl-1-picrylhydrazyl (DPPH) radical, reducing antioxidant power and inhibition of lipid peroxidation antioxidant systems. Oxidative stress was produced in rats by single intraperitoneal injection of 150 mg/kg alloxan and serum concentration of malonaldehyde (MDA), superoxide dismutase (SOD) and catalase (CAT) were determined. Five experimental groups of rats (n=6) were used for the study. Two groups of diabetic rats received oral daily doses of 100 and 200 mg/kg Justicia carnea leaf extract respectively while gilbenclamide (5 mg/ml); a standard diabetic drug was also given to a specific group for 14 days. From the result, the leaf extract contained a higher concentration of flavonoids followed byphenols, ascorbic acid, lycopene and β-carotene. The extract displayed more potent reducing power ability with EC50 of 40 µg/ml compared to BHA (EC50 of 400µg/ml). The percentage DPPH radical scavenging activity of the extract was also higher with EC50 of 200µg/ml and increased with increase in concentration while BHA had EC50of 320µg/ml. The inhibition of lipid peroxidation also increased with increase in concentration with EC50 of 58µg/ml and comparable with BHA (EC50=60µg/ml). The effect of the plant extract on antioxidant enzyme activities was concentration-dependent. Administration of 100mg/kg of the plant extract resulted in a significant decrease (p<0.05) in serum MDA concentration, while 200 mg/kg of the extract caused a significant (p˂0.05) increase in superoxide dismutase (SOD) and catalase activities with a non-significant increase (p>0.05) in the serum level of MDA when compared with the diabetic untreated group. These findings suggest that ethanol leaf extract of Justicia carnea have antioxidant properties and could handle diabetes-induced oxidative stress.
Keywords:
- Justicia carnea
- antioxidant activity
- oxidative stress
- alloxan-induced
- diabetes
- superoxide dismutase
- catalase
- MDA
How to Cite
Chidi, U. S., Nnenna, A. O., Kelechi, A. K., Chijindu, M. F., & Nebolisa, O. C. (2020). In-vitro and In-vivo Antioxidant Activity of Ethanol Leaf Extract of Justicia carnea. International Journal of Biochemistry Research & Review, 29(4), 48-60. https://doi.org/10.9734/ijbcrr/2020/v29i430185
References
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Polterat O. antioxidants and free redical scavengers of natural origin. Current Organic Chemistry. 1997;1:415-440.
Gerber M, Rault MCB, Herberg S, Riboli A. Scalbert A, Siess MH. Food and cancer state of the art about the protective effect of fruits and vegetables. Bulletin Cancer. 2002;89:293-312.
Robak J, Marcinkiewiez E. Scavenging of reactive oxygen species as the mechanism of drug action. Polish Journal of Pharmacology. 1995;47:89-98.
Sasaki YF, Kawaguchi S, Kamaya A, Ohshita M, Kabasawa K, Iwama K, Taniguchi K, Tsuda S. The comet assay with 8 mouse organs: results with 39 currently used food additives. Mutation Research/Genetic Toxicology and Environmental Mutagenesis. 2002;519: 103-109.
Vongtau HO, Abbah J, Chindo BA, Mosugu O, Salawu AO, Kwanashie HO, Gamaniel KS. Central inhibitory effects of the methanol extract of Neorautaneniamitis root in rats and mice. Journal of Pharmaceutical Biology. 2005;43:113- 120.
Oluyemi KA, Okwuonu UC, Baxter DG, Oyesola TO. Toxic effects of methanolic extract of Aspiliaafricana leaf on the estrous cycle and uterine tissues of wister rats. International Journal of Morphology. 2007;25:609-614.
Rout SP, Choudary KA, Kar DM, Das L, Jain A. Plants in traditional medical system-future source of new drugs. International Journal of Pharmacy and Pharm Sciences. 2009;1:1-23.
Corrêa GM, Alcântara AFC. Chemical constituents and biological activities of species of Justicia - A review. Brazilian Journal of Pharmacognosy. 2012;22(1): 220-238.
Edward FG. Justicia carnea Jacobina, Flamingo plant. Environmental Horticulture Department; 2014.
Moswa JL, Kapanda N, Mungende DM, Okitolonda W, Mayangi M, Mihigo S, Mbalek. Plants as an important source of Iron for the treatment of Anaemia case of justiciasecunda. NAPRECA Symposium book of proceedings, Antananarivo Madagascar. 2015;11:132-135.
Ani ON, Udedi SC, Akpata EI, Ezeigwe OC, Oguazu CE, Onyishi CK, Nwakaudu EN. Effects of ethanol leaf extract of Justicia carnea on biochemical indices of alloxan-induced diabetic rats. IOSR Journal of Biotechnology and Biochemistry (IOSR-JBB). 2020;6(2):39-46.
Lorke D. A new approach to practical acute toxicity testing. Archives of Toxicology. 1983;54:275-287.
Barros L, Ferreira MJ, Queiros B, Ferreira IC, Batista P. Total phenols, ascorbic acid, á-carotene and lycopene in portuguese wild edible mushrooms and their antioxidant activities, Food Chemistry. 2007;103:1314-419.
Klein BP, Perry AK. Ascorbic acid and vitamin A activity in selected vegetables from different geographical areas of the United States. Journal of Food Science. 1982;47:941-945.
Ebrahimzadeh MA, Seyed MN, Seyed FN, Fatemeh B, Ahmed RB. Antioxidants and free radical scavenging activity of H. officinalis, L. angustiotolius, J. odoratu. B. hyreana and C. speciosum. Pakistan Journal of Pharmceutical Science. 2009; 23(1):29-34.
Barros L, Soraia F, Baptista P, Cristina F, Miguel VB, Isabel CF, Ferreira R. Antioxidant activity of agaricus sp. Muchrooms by chemical biochemical and electrochemical assays. Food Chemistry. 2007;111:61-66.
Sun M. Zigma S. An improved spectrophotometer assay of superoxide dismutase based on epinephrine antioxidation. Analytical Biochemistry. 1978;90:81-89.
Sinha AK. Colorimetric Assay of Catalase. Analytical Biochemistry. 1972;47(2):389-394.
Buege JA, Aust SD. Microsomal Lipid Peroxidation. Methods in Ezymology. 1978;52:302-310.
Orjiakor CA. Studies on the phytochemical and nutritional composition of aqueous leaf extract of Justicia carnea and its effect on some Biochemical parameters in anaemic rats. M.Sc. research project in nutritional Biochemistry; department of Biochemistry, University of Nigeria, Nsukka; 2014.
Onyeabo C, Achi NK, Ekeleme-Egedigwe CA, Ebere CU, Okoro CK. Haematological and biochemical studies on Justicia carnea leaves extract in phenylhydrazine induced-anemia in albino rats. Acta Scientiarum Polonorum Technologia Alimentaria. 2017; 16(2):217–230.
Ajila CM, Naidu KA, Bhat SG, Prasada Rao UJS. Bioactive compounds and antioxidant potential of mango peel extract. Food Chemistry. 2007;105:982–988.
Kumar S, Pandey AK. Chemistry and biological activities of flavonoids: An overview. Scientific World Journal. 2013; 162750.
Saxena M, Saxena J, Pradhan A. Flavonoids and Phenolic acids as antioxidants in plants and human health, International Journal of Pharmaceutical Sciences Review and Research. 2012; 16(2):130-134.
Okwu DE. Phytochemical and vitamin content of indigenous spices of South Eastern Nigeria. Journal of Sustainable Agriculture and the Environment. 2004;6: 30-37.
Okwu, DE, Josiah C. Evaluation of the chemical composition of two Nigerian medicinal plants. African Journal of Biotechnology. 2006;4:357-361.
Reşat A, Kubilay G, Birsen D, Mustafa Ö, Saliha E, Çelik B, BektaşoğluK-Işıl B, Dilek Ö. Comparative evaluation of various total antioxidant capacity assays applied to phenolic compounds with the CUPRAC assay. Molecules. 2007;12:1496-1547.
Hamilton RT, Walsh ME, Remmen HV. Mouse models of oxidative stress indicate a role for modulating healthy aging. Journal of Clinical and Experimental Pathology. 2012;S4:005.
Burton GK, Jauniaux E. Oxidative stress. Best practice and research clinical obstetrics and gynaecology. 2011;25: 287-299.
Matthaus B. Antioxidant activity of extracts obtained from residues of different oil seeds. Journal of Agricultural and Food Chemistry. 2002;50:3444-3452.
Meir SJ, Kanner B, Akiri SPH. Determination and involvement of aqueous reducing compounds in oxidative defense systems of various senescing leaves. Journal of Agricultural and Food Chemistry. 1995;43:1813-1817.
Okawa M, Kinjo J, Nohara T, Ono M. DPPH (1, 1-diphenyl-2-picrylhydrazyl) radical scavenging activity of flavonoids obtained from some medicinal plants. Biological and Pharmaceutical Bulletin. 2001;24:1202-1205.
Zhang J, Li LX, Liu XH, Wang Y, Zhao DB. Flavonoids from Artemisia sphaerocephala and their free readical scavenging activity using DPPH. Chemistry of Natural Compounds. 2012;48:879-880.
Dragan A, Dusanka D, Drago B, Nenad T. Structural-radical scavenging activity relationships of flavonoids. Croatica Chemica Acta. 2003;76:55-61.
Huang D, Ou B, Prior RL. The chemistry behind antioxidants capacity assays. Journal of Agricultural and Food Chemistry. 2005;53:1841-1856.
Ordonez AAL, Gomez JD, Vattuone MA, Isla MI. antioxidant activities of Sechiumedule (Jacq.) Swartz extracts. Food Chemistry. 2006;97:452-458.
Chanda S, Dave R. In vitro models for antioxidant activity evaluation and some medicinal plants possessing antioxidant properties: An overview, African Journal of. Microbiology Research. 2009;3:981-996.
Oktay M, Gülçin I, Küfreviogle ÖI. Determination of in vitro antioxidant activity of fennel (Foeniculum vulgare) seed extracts, LWT-Food Science and Technology. 2003;36:263-271.
Barrera G. Oxidative stress and lipid peroxidation products in cancer progression and therapy. ISRN Oncology. 2012. Available:http://dx.doi.org/10.5402/2012/137289.
Nowak JZ. Oxidative stress, polyunsaturated fatty acids-derived oxidation products and bisretinoids as potential inducer of CNS diseases: focus on age-related macular degeneration. Pharmacological Report. 2014;65:288-304.
Usuki R, Endo Y, Kaneda T. Antioxidant activity of some food plants. Journal of Japan Soc. Food Science and Technology. 1981;28:583.
Gutteridge J.M. Free radicals in disease processes: A compilation of cause and consequence. Free Radical Research. 1993;19:141-158.
Knight JA. Diseases related to oxygen derived free radicals. Annals of Clinical Laboratory Science. 1995;25:111-121.
Khalili RMA, Shafekh SE, Norhayati AA, Fatahudin IM, Rahimah R, Norkamaliah H, Azimah AN. Total phenolic content and in vitro antioxidant activity of winged bean (Psophocarpus tetragonolobus). Pakistan Journal of Nutrition. 2013;12:416-422.
Veeru P, Kishor MP, Meenakshi M. Screening of medicinal plants for antioxidant activities. Journal of Medicinal Plant Research. 2009;3:608-612.
Michalak A. Phenolic compounds and their antioxidant activity in plants growing under heavy metal. Pol J Environ Stud. 2006;15: 523-530.
Lobo V, Patil A, Phatak A, Chandra N. Free radicals, antioxidants and functional foods: Impact on human health. Pharmacognosy Review. 2010;4:118- 126.
Gorus FK, Malaisse WJ, Pipeleers DG. Selective uptake of alloxan by pancreatic B-cells. Biochemical Journal. 1982;208: 513–515.
Weaver DC, McDaniel ML, Lacy PE. Alloxan uptake by isolated rat islets of Langerhans. Endocrinology. 1978;102: 1847–1855.
Lenzen S. The mechanisms of alloxan- and streptozotocin-induced Diabetes. Diabetologia. 2008;51:216-226.
Munday R. Dialuric acid autoxidation. Effects of transition metals on the reaction rate and on the generation of ‘active oxygen’ species. Biochemical Pharmacology. 1988;37:409–413.
Winterbourn CC, Cowden WB, Sutton HC. Auto-oxidation of dialuric acid, divicine and isouramil. Superoxide dependent and independent mechanisms. Biochemical Pharmacology. 1989;38:611–618.
Jacob RA. The Integrated antioxidant system. Nutrition Research. 1995;15:755-766.
Singh HP, Kaur S, Negi K, Kumari S, Saini V, Batish DR. Assessment of in-vitro antioxidant actuivity of essential oil of Eucalyptus citriodora (lemon-scented Eucalypt; Myrtaceae) and its major constituents. LWT Food Science and Technology. 2012;48(2):237–241.
Wohaieb SA, Godin DV. Alteration in free radical tissue defense mechanisms in streptozotocin induced diabetes in rat; effects of insulin treatment. Diabetes. 1987;36:169-173.
Halliwell B, Gutteridge JMC. Free radicals in biology and medicine. 2. Oxford: Clarendon Press; 1989.
Mecord JM, Keele BB, Fridovich I. An enzyme based theory of obligate anaerobiosis, the physiological functions of superoxide dismutase. Proceedings of the National Academy of Sciences of the United States of America. 1976;1024-27.
Chance B, Greenstein DS, Roughton RJW. The mechanisms of catalase action steady state analysis. Archives of Biochemistry Biophysics. 1952;37:301-39.
Thirunavukkarasu SV, Venketaraman S, Lokesh U. In vitro antioxidant and antibacterial activity of polyherbal Manasamitra vatakam drug. Journal of Pharmceutical Research. 2010;3:2042-3047.
Chidambara KN, Singh RP, Jayaprakasha Gk. Antioxidant activities of grape (Vitis Viniveral) pomace extracts. Journal of Agriculture and Food Chemistry. 2002: 505909-5914.
Yeh C, Yen G. Induction of hepatic antioxidant enzymes by phenolic acids in rats is accompanied by increased levels of multidrug resistance-associated protein 3 mRNA Expression. Journal of Nutrition. 2006;136:11-15.
Roland–Gutrerez JM, DoleresLugue de Castro M. The need for better methods for characterization and determination. Trends in Analytical Chemistry. 2007;26:163-170.
Halliwel B, Gutteridge JMC. “Free radicals in biology and medicine.” 4th ed., oxford, UK, Clarendon Press. 2008;48-50.
Farombi EO, Fakoyak A. “Free radical scavenging and antigenotic activities of natural phenolic compounds in dried flowers of hibiscus sabiscussabdariffa”. Molecular Nutrition and Food Research. 2005;49:1120-1128.
Polterat O. antioxidants and free redical scavengers of natural origin. Current Organic Chemistry. 1997;1:415-440.
Gerber M, Rault MCB, Herberg S, Riboli A. Scalbert A, Siess MH. Food and cancer state of the art about the protective effect of fruits and vegetables. Bulletin Cancer. 2002;89:293-312.
Robak J, Marcinkiewiez E. Scavenging of reactive oxygen species as the mechanism of drug action. Polish Journal of Pharmacology. 1995;47:89-98.
Sasaki YF, Kawaguchi S, Kamaya A, Ohshita M, Kabasawa K, Iwama K, Taniguchi K, Tsuda S. The comet assay with 8 mouse organs: results with 39 currently used food additives. Mutation Research/Genetic Toxicology and Environmental Mutagenesis. 2002;519: 103-109.
Vongtau HO, Abbah J, Chindo BA, Mosugu O, Salawu AO, Kwanashie HO, Gamaniel KS. Central inhibitory effects of the methanol extract of Neorautaneniamitis root in rats and mice. Journal of Pharmaceutical Biology. 2005;43:113- 120.
Oluyemi KA, Okwuonu UC, Baxter DG, Oyesola TO. Toxic effects of methanolic extract of Aspiliaafricana leaf on the estrous cycle and uterine tissues of wister rats. International Journal of Morphology. 2007;25:609-614.
Rout SP, Choudary KA, Kar DM, Das L, Jain A. Plants in traditional medical system-future source of new drugs. International Journal of Pharmacy and Pharm Sciences. 2009;1:1-23.
Corrêa GM, Alcântara AFC. Chemical constituents and biological activities of species of Justicia - A review. Brazilian Journal of Pharmacognosy. 2012;22(1): 220-238.
Edward FG. Justicia carnea Jacobina, Flamingo plant. Environmental Horticulture Department; 2014.
Moswa JL, Kapanda N, Mungende DM, Okitolonda W, Mayangi M, Mihigo S, Mbalek. Plants as an important source of Iron for the treatment of Anaemia case of justiciasecunda. NAPRECA Symposium book of proceedings, Antananarivo Madagascar. 2015;11:132-135.
Ani ON, Udedi SC, Akpata EI, Ezeigwe OC, Oguazu CE, Onyishi CK, Nwakaudu EN. Effects of ethanol leaf extract of Justicia carnea on biochemical indices of alloxan-induced diabetic rats. IOSR Journal of Biotechnology and Biochemistry (IOSR-JBB). 2020;6(2):39-46.
Lorke D. A new approach to practical acute toxicity testing. Archives of Toxicology. 1983;54:275-287.
Barros L, Ferreira MJ, Queiros B, Ferreira IC, Batista P. Total phenols, ascorbic acid, á-carotene and lycopene in portuguese wild edible mushrooms and their antioxidant activities, Food Chemistry. 2007;103:1314-419.
Klein BP, Perry AK. Ascorbic acid and vitamin A activity in selected vegetables from different geographical areas of the United States. Journal of Food Science. 1982;47:941-945.
Ebrahimzadeh MA, Seyed MN, Seyed FN, Fatemeh B, Ahmed RB. Antioxidants and free radical scavenging activity of H. officinalis, L. angustiotolius, J. odoratu. B. hyreana and C. speciosum. Pakistan Journal of Pharmceutical Science. 2009; 23(1):29-34.
Barros L, Soraia F, Baptista P, Cristina F, Miguel VB, Isabel CF, Ferreira R. Antioxidant activity of agaricus sp. Muchrooms by chemical biochemical and electrochemical assays. Food Chemistry. 2007;111:61-66.
Sun M. Zigma S. An improved spectrophotometer assay of superoxide dismutase based on epinephrine antioxidation. Analytical Biochemistry. 1978;90:81-89.
Sinha AK. Colorimetric Assay of Catalase. Analytical Biochemistry. 1972;47(2):389-394.
Buege JA, Aust SD. Microsomal Lipid Peroxidation. Methods in Ezymology. 1978;52:302-310.
Orjiakor CA. Studies on the phytochemical and nutritional composition of aqueous leaf extract of Justicia carnea and its effect on some Biochemical parameters in anaemic rats. M.Sc. research project in nutritional Biochemistry; department of Biochemistry, University of Nigeria, Nsukka; 2014.
Onyeabo C, Achi NK, Ekeleme-Egedigwe CA, Ebere CU, Okoro CK. Haematological and biochemical studies on Justicia carnea leaves extract in phenylhydrazine induced-anemia in albino rats. Acta Scientiarum Polonorum Technologia Alimentaria. 2017; 16(2):217–230.
Ajila CM, Naidu KA, Bhat SG, Prasada Rao UJS. Bioactive compounds and antioxidant potential of mango peel extract. Food Chemistry. 2007;105:982–988.
Kumar S, Pandey AK. Chemistry and biological activities of flavonoids: An overview. Scientific World Journal. 2013; 162750.
Saxena M, Saxena J, Pradhan A. Flavonoids and Phenolic acids as antioxidants in plants and human health, International Journal of Pharmaceutical Sciences Review and Research. 2012; 16(2):130-134.
Okwu DE. Phytochemical and vitamin content of indigenous spices of South Eastern Nigeria. Journal of Sustainable Agriculture and the Environment. 2004;6: 30-37.
Okwu, DE, Josiah C. Evaluation of the chemical composition of two Nigerian medicinal plants. African Journal of Biotechnology. 2006;4:357-361.
Reşat A, Kubilay G, Birsen D, Mustafa Ö, Saliha E, Çelik B, BektaşoğluK-Işıl B, Dilek Ö. Comparative evaluation of various total antioxidant capacity assays applied to phenolic compounds with the CUPRAC assay. Molecules. 2007;12:1496-1547.
Hamilton RT, Walsh ME, Remmen HV. Mouse models of oxidative stress indicate a role for modulating healthy aging. Journal of Clinical and Experimental Pathology. 2012;S4:005.
Burton GK, Jauniaux E. Oxidative stress. Best practice and research clinical obstetrics and gynaecology. 2011;25: 287-299.
Matthaus B. Antioxidant activity of extracts obtained from residues of different oil seeds. Journal of Agricultural and Food Chemistry. 2002;50:3444-3452.
Meir SJ, Kanner B, Akiri SPH. Determination and involvement of aqueous reducing compounds in oxidative defense systems of various senescing leaves. Journal of Agricultural and Food Chemistry. 1995;43:1813-1817.
Okawa M, Kinjo J, Nohara T, Ono M. DPPH (1, 1-diphenyl-2-picrylhydrazyl) radical scavenging activity of flavonoids obtained from some medicinal plants. Biological and Pharmaceutical Bulletin. 2001;24:1202-1205.
Zhang J, Li LX, Liu XH, Wang Y, Zhao DB. Flavonoids from Artemisia sphaerocephala and their free readical scavenging activity using DPPH. Chemistry of Natural Compounds. 2012;48:879-880.
Dragan A, Dusanka D, Drago B, Nenad T. Structural-radical scavenging activity relationships of flavonoids. Croatica Chemica Acta. 2003;76:55-61.
Huang D, Ou B, Prior RL. The chemistry behind antioxidants capacity assays. Journal of Agricultural and Food Chemistry. 2005;53:1841-1856.
Ordonez AAL, Gomez JD, Vattuone MA, Isla MI. antioxidant activities of Sechiumedule (Jacq.) Swartz extracts. Food Chemistry. 2006;97:452-458.
Chanda S, Dave R. In vitro models for antioxidant activity evaluation and some medicinal plants possessing antioxidant properties: An overview, African Journal of. Microbiology Research. 2009;3:981-996.
Oktay M, Gülçin I, Küfreviogle ÖI. Determination of in vitro antioxidant activity of fennel (Foeniculum vulgare) seed extracts, LWT-Food Science and Technology. 2003;36:263-271.
Barrera G. Oxidative stress and lipid peroxidation products in cancer progression and therapy. ISRN Oncology. 2012. Available:http://dx.doi.org/10.5402/2012/137289.
Nowak JZ. Oxidative stress, polyunsaturated fatty acids-derived oxidation products and bisretinoids as potential inducer of CNS diseases: focus on age-related macular degeneration. Pharmacological Report. 2014;65:288-304.
Usuki R, Endo Y, Kaneda T. Antioxidant activity of some food plants. Journal of Japan Soc. Food Science and Technology. 1981;28:583.
Gutteridge J.M. Free radicals in disease processes: A compilation of cause and consequence. Free Radical Research. 1993;19:141-158.
Knight JA. Diseases related to oxygen derived free radicals. Annals of Clinical Laboratory Science. 1995;25:111-121.
Khalili RMA, Shafekh SE, Norhayati AA, Fatahudin IM, Rahimah R, Norkamaliah H, Azimah AN. Total phenolic content and in vitro antioxidant activity of winged bean (Psophocarpus tetragonolobus). Pakistan Journal of Nutrition. 2013;12:416-422.
Veeru P, Kishor MP, Meenakshi M. Screening of medicinal plants for antioxidant activities. Journal of Medicinal Plant Research. 2009;3:608-612.
Michalak A. Phenolic compounds and their antioxidant activity in plants growing under heavy metal. Pol J Environ Stud. 2006;15: 523-530.
Lobo V, Patil A, Phatak A, Chandra N. Free radicals, antioxidants and functional foods: Impact on human health. Pharmacognosy Review. 2010;4:118- 126.
Gorus FK, Malaisse WJ, Pipeleers DG. Selective uptake of alloxan by pancreatic B-cells. Biochemical Journal. 1982;208: 513–515.
Weaver DC, McDaniel ML, Lacy PE. Alloxan uptake by isolated rat islets of Langerhans. Endocrinology. 1978;102: 1847–1855.
Lenzen S. The mechanisms of alloxan- and streptozotocin-induced Diabetes. Diabetologia. 2008;51:216-226.
Munday R. Dialuric acid autoxidation. Effects of transition metals on the reaction rate and on the generation of ‘active oxygen’ species. Biochemical Pharmacology. 1988;37:409–413.
Winterbourn CC, Cowden WB, Sutton HC. Auto-oxidation of dialuric acid, divicine and isouramil. Superoxide dependent and independent mechanisms. Biochemical Pharmacology. 1989;38:611–618.
Jacob RA. The Integrated antioxidant system. Nutrition Research. 1995;15:755-766.
Singh HP, Kaur S, Negi K, Kumari S, Saini V, Batish DR. Assessment of in-vitro antioxidant actuivity of essential oil of Eucalyptus citriodora (lemon-scented Eucalypt; Myrtaceae) and its major constituents. LWT Food Science and Technology. 2012;48(2):237–241.
Wohaieb SA, Godin DV. Alteration in free radical tissue defense mechanisms in streptozotocin induced diabetes in rat; effects of insulin treatment. Diabetes. 1987;36:169-173.
Halliwell B, Gutteridge JMC. Free radicals in biology and medicine. 2. Oxford: Clarendon Press; 1989.
Mecord JM, Keele BB, Fridovich I. An enzyme based theory of obligate anaerobiosis, the physiological functions of superoxide dismutase. Proceedings of the National Academy of Sciences of the United States of America. 1976;1024-27.
Chance B, Greenstein DS, Roughton RJW. The mechanisms of catalase action steady state analysis. Archives of Biochemistry Biophysics. 1952;37:301-39.
Thirunavukkarasu SV, Venketaraman S, Lokesh U. In vitro antioxidant and antibacterial activity of polyherbal Manasamitra vatakam drug. Journal of Pharmceutical Research. 2010;3:2042-3047.
Chidambara KN, Singh RP, Jayaprakasha Gk. Antioxidant activities of grape (Vitis Viniveral) pomace extracts. Journal of Agriculture and Food Chemistry. 2002: 505909-5914.
Yeh C, Yen G. Induction of hepatic antioxidant enzymes by phenolic acids in rats is accompanied by increased levels of multidrug resistance-associated protein 3 mRNA Expression. Journal of Nutrition. 2006;136:11-15.
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