Fate of biomolecules during carbon tetrachloride induced oxidative stress and protective nature of Ammoniac baccifera Linn.: A natural antioxidant

G. Lavanya, R. Sivajyothi, M. Manjunath, P. R. Parthasarathy


Ethanol extract of Ammannia baccifera was studied for its protective nature against the oxidative damage of lipids, proteins and DNA in carbon tetrachloride (CCl4)-induced toxicity in rats. CCl4 administration to albino Wistar rats increased the levels of lipid peroxidation, protein carbonyls and decreased the levels of total sulfhydryls. CCl4 also induced the elevation of DNA damage measured by the comet assay. The study revealed that the administration of the ethanol extract of A. baccifera to CCl4 intoxicated rats could significantly (P , 0.01) decrease the levels of lipid peroxidation, protein carbonyls and increased the levels of total sulfhydryls in a dose-dependent manner. It was also found that the ethanol extract of A. baccifera prevent the CCl4-induced elevation of DNA damage in hepatocytes. These results suggest that treatment with the ethanol extract of A. baccifera can minimize the deleterious effects caused by CCl4 through its strong antioxidative and free radical scavenging properties.
Key words: Antioxidants, comet assay, DNA, protein carbonyls, total sulfhydryls

Full Text:



Sohal RS, Weindruch R. Oxidative stress, caloric restriction, and

aging. Science 1996;273:59-63.

Benzie IFF. Antioxidants: Observational epidemiology. In:

Sadler MJ, Strain JJ, Cabellero B, editors. The encyclopedia of

human nutrition. New York: Academic Press; 1999. p. 106-15.

Beckman KB, Ames BN. The free radical theory of aging matures.

Physiol Rev 1998;78:547-81.

Collins AR. Oxidative damage, antioxidants and cancer. Bioassays


Gey KF. Vitamins E plus C and interacting co-nutrients required for

optimal health: A critical and constructive review of epidemiology

and supplementation data regarding cardiovascular disease and

cancer. Biofactors 1998;7:113-74.

Hettiarachchy NS, Glenn KC, Gnanasambandam R, Johnson MG.

Natural antioxidant extracts from fenugreek (Trigonella

foenumgraecum) for ground beef patties. J Food Sci 1996;61:516-9.

Wood LG, Gibson PG, Garg ML. A review of the methodology

for assessing in vivo antioxidant capacity. J Sci Food Agr


Kirtikar KR, Basu BD. Indian medicinal plants. Vol. 2, 2nd ed. In:

Pal Singh BM, editor. New Delhi, India: 1972. p. 1071.

Dhar MN, Dhavan BN, Mehrotra BN, Srimal RC, Tandon JS.

Screening of Indian medicinal plants for biological activities.

Indian J Exp Biol 1973;11:43-54.

Bharathi K, Srinivasan KK. Evaluation of Ammania baccifera

Linn. for antiurolithic activity in albino rats. Indian J Exp Biol


Al-Sharma A, Mitschar LA. Comprehensive surgery of indigenous

Iraqi plants for potential economic value.1. Screening results of

species for alkaloids. J Nat Prod 1979;42:633-42.

Dhanapal R, Kavimani S, Bhyrapur Matha VS, Gupta M,

Basu SK. Antisteroidogenic activity of ethanol extract of

Ammania baccifera (L.) whole plant in female albino mice ovaries.

Iranian J Pharmacol Therapeut 2005;4:43-5.

Shahjahan M, Sabitha KE, Jainu M, Shyamala Devi CS. Effect of

Solanum trilobatum against carbon tetrachloride induced hepatic

damage in albino rats. Indian J Med Res 2004;120:194-8.

Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein

measurement with folin phenol reagent. J Boil Chem


Ohkawa H, Ohishi N, Yagi K. Assay of lipid peroxides in animal

tissue by thiobarbituric acid reaction. Anal Biochem 1979;95:351-8.

Levine RL, Garland D, Oliver CN, Amici A, Climent I, Lenz AG,

et al. Determination of carbonyl content in oxidatively modified

proteins. Met Enzymol 1990;186:464-78.

Sedlack J, Lindsay RH. Estimation of total protein bound and

non-protein bound sulphydryl group in the tissue with Ellman’s

reagent. Anal Biochem 1968;25:7826-33.

Endoh D, Okui T, Ozawa S, Yamato O, Kon Y, Arikawa J, et al.

Protective effect of a lignan-containing flaxseed extract against

CCl4-induced hepatic injury. J Vet Med Sci 2002;64:761-5.

Singh NP, McCoy MT, Tice RR, Schneider EL. A simple technique

for quantitation of low levels of DNA damage in individual cells.

Exp Cell Res 1988;175:184-91.

Kumar R, Das SK, Hossain U, Bhattacharya S. Protective effect

of diphenylmethyl selenocyanate against CCl4-induced hepatic

injury. J Appl Toxicol 2007;27:527-37.

Sebastien G, Fabrice N, ElMoukhtar A, Jean-Marie H, Daniel M.

Assessment of genotoxic effect of benzo[a]pyrene in endotracheally

treated rat using the comet assay. Mutat Res 2003;534:33-43.

Recknagel RO, Glende Jr EA. Carbon tetrachloride hepatotoxicity:

An example of lethal cleavage. CRC Crit Rev Toxicol 1973;2:263-97.

Srivastava SP, Chen NQ, Holtzman JL. The in vitro NADPH

dependent inhibition by CCl4 of the ATP-dependent calcium

uptake of hepatic microsomes from male rats: Studies on the

mechanism of the inactivation of the hepatic microsomal calcium

pump by the CCl3• radical. J Biol Chem 1990;265:8392-9.

Recknagel RO, Glende Jr. EA, Dolak JA, Waller RL. Mechanisms

of carbon tetrachloride toxicity. Pharmacol Ther 1989;43:139-54.

Mansour MA. Protective effects of thymoquinone and

desferrioxamine against hepatotoxicity of carbon tetrachloride in

mice. Life Sci 2000;66:2583-91.

Manna P, Mahau, Sil PC. Aqueous extract of Terminalia arjuna

prevents carbon tetrachloride induced hepatic and renal disorders.

BMC Compl Alter Med 2006;6:33-43.

Davies KJ, Delsignore ME, Lin SW. Protein damage and

degradation by oxygen radicals: II: Modification of amino acids.

J Biol Chem 1987;262:9902-7.

Uchida K, Stadtman ER. Covalent attachment of 4-hydroxynonenal

to glyceraldehydes 3-phosphate dehydrogenase: A possible

involvement of intra- and intermolecular cross-linking reaction.

J Biol Chem 1993;268:6388-93.

Lim PS, Cheng YM, Wei YH. Increase in oxidative damage to lipids

and proteins in skeletal muscle of uremic patients. Free Radiat Res


Sarkar A, Sreenivasan Y, Govindarajan T, Ramesh K, Manna SK.

b-D-Glucoside suppresses tumor necrosis factor-induced

activation of nuclear transcription factor NkB but potentiates

apoptosis. J Biol Chem 2004;279:33768-81.

Ahmad S, Singh V, Rao GS. Antioxidant potential in serum

and liver of albino rats exposed to benzene. Indian J Exp Biol


Yildiz D, Dalkilic S, Yildiz H, Oztas H. Methyl parathione-induced

changes in free and protein-bound SH levels in rat tissues. Toxicol

Mechan Met 2006;16:347-52.

Ross D. Glutathione, free radicals and chemotherapeutic agents:

Mechanics of free radical induced toxicity and glutathione‑dependent

protection. Pharmac Ther 1988;37:231-49.

Raja S, Nazeer Ahamed KF, Kumar V, Mukherjee K,

Bandyopadhyay A, Mukherjee PK. Antioxidant effect of Cytisus

scoparius against carbon tetrachloride treated liver injury in rats.

J Ethnopharmacol 2007;109:41-7.

Eastman A, Barry MA. The origins of DNA breaks: A consequence

of DNA damage, DNA repair, or apoptosis? Cancer Invest


Møller P, Wallin H. Adduct formation, mutagenesis and nucleotide

excision repair of DNA damage produced by reactive oxygen

species and lipid peroxidation product. Mutat Res 1998;410:271-90.

Boll M, Weber LW, Becker E, Stampfl A. Pathogenesis of carbon

tetrachloride-induced hepatocyte injury: Bioactivation of CCl4 by

cytochrome P450 and effects on lipid homeostasis. Z Naturforsch


Sasaki YF, Saga A, Akasaka M, Ishibashi S, Yoshida K, Su YQ,

et al. Detection of in vivo genotoxicity of haloalkanes and

haloalkenes carcinogenic to rodents by the alkaline single cell gel

electrophoresis (comet) assay in multiple mouse organs. Mutat

Res 1998;419:13-20.

DOI: http://dx.doi.org/10.22377/ijgp.v3i4.105


  • There are currently no refbacks.