In vitro protection of biological macromolecules against oxidative stress and in vivo toxicity evaluation of Acacia nilotica (L.) and ethyl gallate in rats
© Mohan et al.; licensee BioMed Central Ltd. 2014
Received: 5 May 2014
Accepted: 15 July 2014
Published: 21 July 2014
Recently, enormous research has been focused on natural bioactive compounds possessing potential antioxidant and anticancer properties using cell lines and animal models. Acacia nilotica (L.) is widely distributed in Asia, Africa, Australia and Kenya. The plant is traditionally used to treat mouth, ear and bone cancer. However, reports on Acacia nilotica (L.) Wild. Ex. Delile subsp. indica (Benth.) Brenan regarding its toxicity profile is limited. Hence in this study, we investigated the antioxidant capacity and acute toxicity of ethyl gallate, a phenolic antioxidant present in the A. nilotica (L.) leaf extract.
The antioxidant activity of ethyl gallate against Fenton’s system (Fe3+/H2O2/ascorbic acid) generated oxidative damage to pBR322 DNA and BSA was investigated. We also studied the interaction of ethyl gallate to CT-DNA by wave scan and FTIR analysis. The amount of ethyl gallate present in the A. nilotica (L.) leaf extract was calculated using HPLC and represented in gram equivalence of ethyl gallate. The acute toxicity profile of ethyl gallate in the A. nilotica (L.) leaf extract was analyzed in albino Wistar rats. Measurement of liver and kidney function markers, total proteins and glucose were determined in the serum. Statistical analysis was done using statistical package for social sciences (SPSS) tool version 16.0.
Ethyl gallate was found to be effective at 100 μg/mL concentration by inhibiting the free radical mediated damage to BSA and pBR322 DNA. We also found that the interaction of ethyl gallate and A. nilotica (L.) leaf extract to CT-DNA occurs through intercalation. One gram of A. nilotica (L.) leaf extract was found to be equivalent to 20 mg of ethyl gallate through HPLC analysis. Based on the acute toxicity results, A. nilotica (L.) leaf extract and ethyl gallate as well was found to be non-toxic and safe.
Results revealed no mortality or abnormal biochemical changes in vivo and the protective effect of A. nilotica (L.) leaf extract and ethyl gallate on DNA and protein against oxidative stress in vitro. Hence, A. nilotica (L.) leaf extract or ethyl gallate could be used as potential antioxidants with safe therapeutic application in cancer chemotherapy.
KeywordsAcacia nilotica (L.) Ethyl gallate Antioxidant Oxidative damage Toxicity DNA interaction
Oxidative stress due to reactive oxygen species/reactive nitrogen species (ROS/RNS) is associated with many diseases like cancer, cardiovascular disease, diabetes, arthritis and neurodegeneration . ROS generation damages the biological macromolecules such as proteins, DNA, polysaccharides and lipids. These free radicals are scavenged by compounds that are capable of donating a hydrogen atom or by activating the antioxidant enzymes . Many synthetic antioxidants were produced to protect the biological macromolecules against oxidative damage. However, there has been an increasing concern regarding the toxic effect of the available antioxidants. Therefore, in recent years, the interest in natural substances from medicinal plants has increased enormously due to the alleviation of diseases by the neutralization of bio-macromolecular oxidation .
Acacia nilotica (L.) belonging to the family Fabaceae and sub-family Mimosoideae is a medium sized tree with a variety of medicinal uses. The species is widely distributed in various tropical and sub-tropical countries around the world. The plant is used traditionally in several parts of Chhattisgarh state of India for treating cancers of the mouth, bones and skin, and in West Africa for tumors of ears, eyes, or testicles . Twigs of the plant are used as tooth brushes in many parts of India and Africa [5, 6]. Various scientific reports suggest that A. nilotica (L.) is rich in bioactive compounds and nutrients to treat many diseases such as cold, bronchitis, asthma, diabetes, diarrhoea, dysentery, blindness, bleeding piles and leucoderma to name a few [7, 8]. A. nilotica (L.) has been reported to exhibit potent antioxidant activity as well, and it has been shown to be significant in comparison with quercetin, tocopherol, ascorbic acid and catechin. Leaves of A. nilotica (L.) are found to be rich in rutin and apigenin-6, 8-bi's-C-β-D-glucopyranoside [4, 9]. Kalaivani et al. [10, 11] has reported the antioxidant capacity of ethyl gallate isolated from the ethanol leaf extract of Acacia nilotica (L.) Wild. ex. Delile subsp. indica (Benth.) Brenan. Subsequently, the cytotoxic activity of ethyl gallate and A. nilotica (L.) leaf extract on HeLa cancer cells and Vero normal cells were established. Furthermore, ethyl gallate isolated from Galla rhois was reported to possess anticancer activity against human leukemia cell line through induction of apoptosis .
Ethyl gallate is a phenolic antioxidant compound present in foods with cancer prevention potential . Therefore, the medicinal and protective properties of ethyl gallate isolated from A. nilotica (L.) leaf extract could be employed in the formulation of drugs. However, there is a paucity of literature information on the toxicity profile of A. nilotica (L.) leaf extract or ethyl gallate. The knowledge on their toxicity profile is essential in order to develop a potent drug from them. Thus, the objective was to determine the safety of A. nilotica (L.) leaf extract and ethyl gallate by assessing their dose-dependent toxicity profile on female albino Wistar rats. The attenuation of DNA or protein damage by Fenton’s system-generated free radicals was also evaluated. In addition, the nature of interaction of A. nilotica (L.) leaf extract and ethyl gallate on CT-DNA was examined by FTIR spectroscopy.
The chemicals ethyl gallate, bovine serum albumin (BSA) and Calf thymus DNA (CT-DNA) were purchased from Sigma-Aldrich Chemical Co (St Louis, MO. USA). pBR322 DNA was purchased from GENEI (Bangalore, India). Biochemical kits were obtained from Span diagnostics (Surat, Gujarat, India), and all other reagents used were of analytical grade.
Plant material and extraction
Leaves of Acacia nilotica (L.) Wild. ex. Delile subsp. indica (Benth.) Brenan were collected from Vellore district, Tamilnadu, India and identified by Dr. G.V.S. Murthy, Scientist-in-Charge, Botanical Survey of India, Southern Regional Centre, Tamilnadu Agricultural University, Coimbatore, India (Voucher number: 1035). A. nilotica (L.) dried leaves (100 g) were extracted exhaustively using ethanol by Soxhlet extraction yielding 40 g after evaporation as described in our previous report . In addition, for validation of the presence of ethyl gallate in the leaves of A. nilotica (L.), 1 g of leaves were extracted by maceration with 20 mL of different solvents separately like methanol-acetonitrile-10 mM ammonium acetate containing 0.1% formic acid (10:25:65 v/v/v) according to the method of Gao et al. , ethanol and water. The supernatant was recovered after filtering the extract through Whatman No. 1 filter paper. The solvent present was evaporated in a vacuum rotary evaporator to obtain the dry extract. The dried extracts were dissolved in methanol-acetonitrile-10 mM ammonium acetate containing 0.1% formic acid (10:25:65 v/v/v) (1.0 mg/mL) and filtered through sterile 0.22 μm millipore filter and subjected to HPLC analysis.
Protein damage assay
The protective effect of A. nilotica (L.) leaf extract and ethyl gallate were tested against Fenton’s system generated protein oxidation by the electrophoretic pattern on Sodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) according to the method of Wang et al.  with slight modification. Reaction mixture containing 1 mg/mL of bovine serum albumin (BSA), Ferric chloride (FeCl3) (50 μM), hydrogen peroxide (H2O2) (1 mM) and ascorbic acid (100 μM) with or without A. nilotica (L.) leaf extract (100 μg/mL), ethyl gallate (100 μg/mL) or butylated hydroxyl toluene (BHT) (100 μg/mL) was made up to 1.2 mL in 20 mM potassium phosphate buffer of pH 7.4. The reaction mixture was incubated at 37°C for 3 h. Electrophoresis of the samples were carried out according to Laemmli’s method  in 12% SDS-PAGE and analyzed using the gel documentation system, AlphaImager HP, Cell Biosciences (Santa Clara, CA).
DNA damage assay
The protective effect of A. nilotica (L.) leaf extract and ethyl gallate were tested for deoxyribonucleic acid (DNA) damage based on the method of Lee et al. . The pBR322 plasmid DNA (200 ng) was oxidized using Fenton’s system (Fe3+/H2O2/ascorbic acid) in the presence or absence of A. nilotica (L.) leaf extract or ethyl gallate or quercetin at 100 μg/mL concentration for 30 min at 37°C. After incubation of these compounds with DNA, 5 μL of all the samples were loaded along with gel loading dye in 1% agarose gel for electrophoresis. The gel was scanned using the gel documentation system, AlphaImager HP, Cell Biosciences (Santa Clara, CA).
DNA interaction by FTIR and UV analysis
FTIR spectroscopy is widely used in recent years for the interaction studies of DNA with natural compounds. A solution of CT-DNA was made with 10 mM Tris-HCl buffer of pH 7.4 and its purity was verified by its absorbance at 260 and 280 nm. Different concentrations of DNA (0.1 to 1 mg mL-1) were analyzed for its binding capacity with a single concentration of A. nilotica (L.) leaf extract or ethyl gallate .
IRAffinity-1 FTIR spectrophotometer (Shimadzu, Columbia, Maryland, USA) was used for recording the spectra using DTGS detector, Ni-Chrome source and KBr beam splitter. One hundred scans for each sample with 4 cm-1 resolution were recorded and evaluated using OMNIC software. The interaction of A. nilotica (L.) leaf extract and ethyl gallate with CT-DNA was evaluated by comparing the shift in the spectrum formed individually or as complexes. A UV-Visible spectrum was also recorded by wave scan range from 200 to 800 nm using Systronics AU-2701 UV-Vis double beam spectrophotometer (Gujarat, India).
A stock of 1 mg/mL of the A. nilotica (L.) leaf extract was prepared using 0.1% ethanol. From this, 100 μL was taken and diluted to 3 mL with methanol. Wave scan analysis was carried out using Systronics AU-2701 UV-Vis double beam spectrophotometer in the wave length ranging from 200 to 800 nm. The peak obtained was compared with that of ethyl gallate. Following this, A. nilotica (L.) leaf extract was dissolved in HPLC grade methanol at a concentration of 1.0 mg/mL, filtered through 0.22 μm filter and subjected to HPLC Schimadzu isocratic system equipped with Luna C18 column. Separation was achieved using acetonitrile/water and the peak obtained was compared with the pure ethyl gallate as standard at 272 nm.
Validation of the presence of ethyl gallate in A. nilotica (L.) leaf extract with different solvents were also done using YOUNGLIN HPLC instrument Acme 9000 with vacuum degasser and mixer. The instrument was equipped with a gradient pump SP930D, a UV/Vis detector UV730D and a Kromasil 100-5C18 column with a length of 250 x 4.6 mm. Data was integrated by the software YOUNGLIN Autochro-3000 chromatograph data system. Separation was achieved by isocratic mobile phase consisting of methanol-acetonitrile-10 mM ammonium acetate containing 0.1% formic acid (10:25:65, v/v/v) with a flow rate of 0.5 mL/min. Peak area of the sample was compared with that of the standard at 291 nm.
Forty eight female albino Wistar rats of six to eight weeks old were obtained from the Institutional animal house, VIT University, Vellore, Tamilnadu, India. All animals were maintained under standard conditions of temperature (28 ± 2°C) and light (12 h light/dark cycles). The animals were housed in polypropylene cages (45 × 24 × 15 cm) and fed with standard diet pellets and water ad libitum. Animals were handled according to the University and Institutional Regulations, administered by the Animal Ethical Committee, VIT University. The protocols performed on the animals were approved and conducted in accordance with the National Institute of Health Guide (VIT/IAEC/V/017/2012).
Acute toxicity study
Rats were divided into eight groups based on their body weights. Animals were deprived of food but not water, 15 h prior to the administration of test substances. Toxicity was assessed by oral administration of 1 mL of A. nilotica (L.) leaf extract and ethyl gallate by gavage feeding and monitored for any mortality up to 14 days. Group 1 rats served as control receiving 1.0 mL of the vehicle (0.1% ethanol); Group 2 rats received A. nilotica (L.) leaf extract (250 mg/kg body weight, equivalent to 5 mg/kg ethyl gallate); Group 3 rats received A. nilotica (L.) leaf extract (500 mg/kg body weight, equivalent to 10 mg/kg ethyl gallate); Group 4 rats received A. nilotica (L.) leaf extract (1000 mg/kg body weight, equivalent to 20 mg/kg ethyl gallate); Group 5 rats received A. nilotica (L.) leaf extract (2000 mg/kg body weight); Group 6 rats received ethyl gallate (5 mg/kg body weight); Group 7 rats received ethyl gallate (10 mg/kg body weight); Group 8 rats received ethyl gallate (20 mg/kg body weight).
Body weights were recorded on 0th and 14th day for each group and all rats were decapitated after an overnight fast . Liver and kidney tissues were removed and rinsed with saline solution, observed for any lesions and weighed. Blood was collected for the assessment of total serum proteins, glucose, liver and kidney function markers such as aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) activities; and total bilirubin and creatinine levels in the serum using Span diagnostic reagent kits . Serum was separated from the collected blood after centrifugation at 3000 rpm for 10 min without any anticoagulant.
All quantitative measurements were expressed as mean ± standard error (SEM). Statistical analysis was performed by one way analysis of variance (ANOVA) followed by Duncan’s Multiple Range Test (DMRT) using Statistical Package for Social Sciences (SPSS) Version 16.0. p < 0.05 were considered statistically significant.
Results and discussion
Although medicinal plants and the plant derived bioactive compounds appear to contribute to the prevention and progression of diseases, they are not nutrients. Hence, the knowledge of their safety and side-effects, if any, are important considerations for their effective use in disease management. In this regard, the general toxicity of medicinal plants and their derivatives needs to be validated from a toxicological point of view. In our previous report, we have determined the titres of phenolics in the A. nilotica (L.) leaf extract and assessed their antioxidant activity . Ethyl gallate, a major phenolic compound obtained from A. nilotica (L.) leaf extract has also been shown to inhibit the growth of cancer cells in vitro. In this report, A. nilotica (L.) leaf extract and ethyl gallate was evaluated for their protection against Fenton’s system generated free radical damage on biological macromolecules like DNA and protein. The nature of interaction of A. nilotica (L.) leaf extract and ethyl gallate to CT-DNA was analyzed using FTIR and UV spectroscopy. The toxicological profile was also studied using albino Wistar rats.
Protection of BSA and DNA damage against oxidative stress
Hydroxyl radicals are well known cause for damaging biological macromolecules leading to mutation, cancer and age-related diseases . Polyphenols are known to inhibit the adverse effects of oxidative stress through their anticancer and antimutagenic properties . The results of this study show that A. nilotica (L.) leaf extract and ethyl gallate act as active scavengers of hydroxyl radicals, thereby protecting BSA and pBR322 DNA, without causing any toxicity. These results are in agreement with the previous work of Singh et al.  using A. nilotica (L.) green pod extracts against DNA damage.
FTIR and wave scan analysis
Acute toxicity study in rats
An acute toxicity study was performed to evaluate the safe administrable doses of A. nilotica (L.) leaf extract or ethyl gallate in female albino Wistar rats for 14 days. Generally, any test substance showing an LD50 of 1000 mg/kg through oral mode can be considered safe . Group 1 received the vehicle alone (0.1% ethanol); groups 2 to 4 received A. nilotica (L.) leaf extract of 250, 500, 1000 mg/kg body weight (ethyl gallate equivalent of 5, 10 and 20 mg/kg, respectively); group 5 received 2000 mg/kg body weight of A. nilotica (L.) leaf extract or ethyl gallate equivalent of 40 mg/kg; and groups 6 to 8 received ethyl gallate alone at doses of 5, 10 and 20 mg/kg body weight respectively. Administration of these test substances orally did not show any mortality in the treated groups. Hence, the LD50 could not be determined for the administered doses. Previous report shows that A. nilotica (L.) flower extract was found to be non-toxic up to a dose of 2000 mg/kg . Similarly, ethyl acetate fruit extract of A. nilotica (L.) upon oral administration to mice showed LD50 value of 7393.4 mg/kg . In contrast, 20-100% mortality was reported in rats treated with 50-500 mg/kg A. nilotica (L.) fruit extract on intraperitoneal administration . However, in the present investigation no mortality was observed for A. nilotica (L.) leaf extract on oral administration to rats.
Effect of A. nilotica (L.) leaf extract and ethyl gallate on weight analysis in rats
Body weight gain (g)
Liver weight (g)
Kidney weight (g)
10.00 ± 1.0a
5.44 ± 0.16a,b
1.28 ± 0.06a
15.33 ± 1.52c,d
5.29 ± 0.39a
1.27 ± 0.18a
21.66 ± 1.15e
6.76 ± 0.43b,c
1.49 ± 0.12a,b
12.00 ± 1.73a,b
7.01 ± 0.95c
1.63 ± 0.06b
17.66 ± 2.30c,d
8.82 ± 0.97d
2.85 ± 0.15c
18.66 ± 1.15c
7.49 ± 0.86c
2.66 ± 0.16c
12.66 ± 1.55b
7.11 ± 0.97c
1.47 ± 0.09a,b
14.33 ± 1.15b,c
6.75 ± 0.81b,c
1.42 ± 0.19a,b
No significant difference in the serum total protein, albumin, globulin and glucose was found between the rats fed with A. nilotica (L.) leaf extract on ethyl gallate equivalent basis and those fed with ethyl gallate alone. Significant differences in total bilirubin level, however, existed between the rats that received A. nilotica (L.) leaf extract, 500 mg/kg body weight (ethyl gallate equivalent of 10 mg/kg, 0.34 ± 0.01 mg/dL) and those receiving 10 mg/kg body weight of ethyl gallate (0.26 ± 0.01 mg/dL). The creatinine levels in the groups fed with A. nilotica (L.) leaf extract (group 2, 250 mg/kg body weight, 0.44 ± 0.02 mg/dL; and group 3, 500 mg/kg body weight, 0.51 ± 0.02 mg/dL) were significantly different (at 95% confidence) from ethyl gallate fed groups (group 6, 5 mg/kg body weight, 0.54 ± 0.02; and group 7, 10 mg/kg body weight, 0.38 ± 0.01 mg/dL).
No significant difference was found for AST between the groups receiving A. nilotica (L.) leaf extract (in ethyl gallate equivalence) and those receiving ethyl gallate. Likewise, no significant difference was found between any groups for ALP except for the groups treated with 250 mg/kg body weight (92.54 ± 4.27 U/L) of A. nilotica (L.) leaf extract (equivalent to 5 mg/kg ethyl gallate) and 5 mg/kg body weight (57 ± 2.63 U/L) of ethyl gallate at p < 0.05. However, significant difference was found for ALT between groups fed with 500 and 1000 mg/kg body weight of A. nilotica (L.) leaf extract (26.52 ± 1.23 and 30.05 ± 1.38 U/L) and 10 and 20 mg/kg of ethyl gallate (20.50 ± 0.94 and 24.67 ± 1.13 U/L).
Taken together, the liver and kidney function markers in the serum was found to be elevated with increase in the concentration of the test substances. When the liver marker values were compared on an equivalent basis of ethyl gallate, there were differences in liver markers between A. nilotica (L.) leaf extract and ethyl gallate, which could be attributed due to the presence of other constituents present in the A. nilotica (L.) leaf extract. Nevertheless, those changes do not appear to have any consequential clinical significance, since the biochemical indicators were found to be within the normal ranges . Our results are also in line with the report by Kannan et al. that the methanolic extract of the aerial part of A. nilotica (L.) offers protection against hepatotoxicity induced by acetaminophen in Wistar rats.
A. nilotica (L.) leaf extract or ethyl gallate does not appear to possess any toxicity in vivo as evidenced by zero mortality, suggesting that a wide margin of safety is possible for the selected therapeutic doses. Secondly, biochemical changes in the serum did not show any signs of toxicity, indicating their safety. In addition, the absence of DNA or protein damage indicates that the test substances are effective antioxidants against the hydroxyl radicals generated by the Fenton’s system. FTIR and UV-Vis spectral results demonstrate, for the first time, that A. nilotica (L.) leaf extract and ethyl gallate has a binding affinity to DNA by intercalation. The mode of interaction of these substances to CT-DNA and the protection offered to pBR322 DNA and BSA suggests their potential for their use in cancer chemotherapy. Further work would involve the evaluation of the absorption and bioavailability of ethyl gallate and other constituents in the A. nilotica (L.) leaf extract through oral ingestion in rats.
Fourier transform infrared
High performance liquid chromatography
Reactive oxygen species
Reactive nitrogen species
Calf-thymus deoxyribonucleic acid
Sodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis
Butylated hydroxyl toluene
- FeCl3 :
- H2O2 :
Bovine serum albumin
- LD50 :
Lethal dose 50
Revolutions per minute
Analysis of variance
Duncan’s multiple range test
Statistical package for social sciences.
The authors are thankful to the VIT University for providing the infrastructure, constant support and encouragement. We also thank the School of Advanced Sciences (SAS) for performing FTIR analysis.
- Donne DI, Rossi R, Colombo R, Giustarini D, Milzani A: Biomarkers of oxidative damage in human disease. Clin Chem. 2006, 52: 601-623.View ArticleGoogle Scholar
- Oboh G: Antioxidant properties of some commonly consumed and underutilized tropical legumes. Eur Food Res Tech. 2006, 224: 61-65.View ArticleGoogle Scholar
- Silva BM, Andrade PB, Valentaoo P, Ferreres F, Seabra RM, Ferreira MA: Quince (Cydonia oblonga Miller) fruit (pulp, peel and seed) and jam: antioxidant activity. J Agric Food Chem. 2004, 52: 4705-4712.View ArticlePubMedGoogle Scholar
- Kalaivani T, Mathew L: Free radical scavenging activity from leaves of Acacia nilotica (L.) Wild. ex Delile, an Indian medicinal tree. Food Chem Toxicol. 2010, 48: 298-305.View ArticlePubMedGoogle Scholar
- Hooda A, Rathee M, Singh J: Chewing sticks in the era of toothbrush: a review. Internet J Fam Pract. 2009, 9: 1-Google Scholar
- Jansen PCM, Cardon D: Dyes and tannins. Plant Resources of Tropical Africa 3. PROTA Foundation. 2005, Wageningen, Netherlands: Backhuys Publishers, 19-25.Google Scholar
- Ambasta SP: The Wealth of India. A dictionary of Indian raw materials and industrial products. 1986, New Delhi: The useful plants of India, CSIR, 13: 198-Google Scholar
- Ali A, Akhtar N, Khan BA, Khan MA, Rasul A, Zaman SU, Khalid N, Waseem K, Mahmood T, Ali L: Acacia nilotica: a plant of multipurpose medicinal uses. J Med Plants Res. 2012, 6: 1492-1496.Google Scholar
- Seigler DS: Phytochemistry of Acacia-sensu lato. Biochem Syst Ecol. 2003, 31: 845-873.View ArticleGoogle Scholar
- Kalaivani T, Rajasekaran C, Mathew L: Free radical scavenging, cytotoxic, and hemolytic activities of an active antioxidant compound ethyl gallate from leaves of Acacia nilotica (L.) wild. Ex. Delile subsp. Indica (Benth.) Brenan. J Food Sci. 2011, 76: 144-149.View ArticleGoogle Scholar
- Kalaivani T, Rajasekaran C, Suthindhiran K, Mathew L: Free radical scavenging, cytotoxic and haemolytic activities from leaves of Acacia nilotica (L.) Wild. ex. Delile subsp. indica (Benth.) Brenan. Evid Based Complement Alternat Med. 2010, 2010: 1-8.Google Scholar
- Kim WH, Song HO, Choi HJ, Bang HI, Choi DY, Park H: Ethyl gallate induces apoptosis of HL-60 cells by promoting the expression of caspases-8,-9,-3, apoptosis-inducing factor and endonuclease G. Int J Mol Sci. 2012, 13: 11912-11922.View ArticlePubMedPubMed CentralGoogle Scholar
- Gotes J, Kasian K, Jacobs H, Cheng ZQ, Mink SN: Benefits of ethyl gallate versus norepinephrine in the treatment of cardiovascular collapse in Pseudomonas aeruginosa septic shock in dogs. Crit Care Med. 2012, 40: 560-572.View ArticlePubMedGoogle Scholar
- Gao S, Zhan Q, Li J, Yang Q, Li X, Chen W, Sun L: LC-MS/MS method for simultaneous determination of ethyl gallate and its major metabolite in rat plasma. Biomed Chromatogr. 2010, 24: 472-478.PubMedGoogle Scholar
- Wang BS, Lin SS, Hsiao WC, Fan JJ, Fuh LF, Duh PD: Protective effects of aqueous extract of Welsh onion green leaves on oxidative damage of reactive oxygen and nitrogen species. Food Chem. 2006, 98: 149-157.View ArticleGoogle Scholar
- LaemmLi UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970, 227: 680-685.View ArticlePubMedGoogle Scholar
- Lee JC, Kim HR, Kim J, Jang YS: Antioxidant property of an ethanol extract of the stem of Opuntia ficus-indica var. Saboten. J Agri Food Chem. 2002, 50: 6490-6496.View ArticleGoogle Scholar
- Ghosh P, Devi PG, Priya R, Amrita A, Sivaramakrishna A, Babu S, Siva R: Spectroscopic and in silico evaluation of interaction of DNA with six anthraquinone derivatives. Appl Biochem Biotechnol. 2013, 170: 1127-1137.View ArticlePubMedGoogle Scholar
- Armed Forces Institute of Pathology: laboratory methods in Histotechnology. Edited by: Prophet EB, Mills B, Arrington JB, Sobin LH. 1992, Washington DC: American Registry of Pathology, 25-53.Google Scholar
- Sharma P, Shankar S, Agarwal A, Singh R: Variation in serum lipids and liver function markers in lindane exposed female wistar rats: attenuating effect of curcumin, vitamin C and vitamin E. Asian J Exp Biol Sci. 2010, 1: 440-444.Google Scholar
- Prakash D, Singh BN, Upadhyay G: Antioxidant and free radical scavenging activities of phenols from onion. Food Chem. 2007, 102: 1389-1393.View ArticleGoogle Scholar
- Singh BN, Singh BR, Singh RL, Prakash D, Sarma BK, Singh HB: Antioxidant and anti-quorum sensing activities of green pod of Acacia nilotica L. Food Chem Toxicol. 2009, 47: 778-786.View ArticlePubMedGoogle Scholar
- Teel RW: Ellagic acid binding to DNA as a possible mechanism for its antimutagenic and anticarcinogenic action. Can Lett. 1986, 30: 329-336.View ArticleGoogle Scholar
- Boubaker J, Mansour HB, Ghedira K, Ghedira LC: Polar extracts from (Tunisian) Acacia salicina Lindl. Study of the antimicrobial and antigenotoxic activities. BMC Complemen Altern Med. 2012, 12: 37-View ArticleGoogle Scholar
- Ghomi M, Letellier R, Liquier J, Taillandier E: Interpretation of DNA vibrational spectra by normal coordinate analysis. Int J Biochem. 1990, 22: 691-699.View ArticlePubMedGoogle Scholar
- Tyagi G, Jangir DK, Singh P, Mehrotra R: DNA interaction studies of an anticancer plant alkaloid, vincristine, using fourier transform infrared spectroscopy. DNA and Cell Biol. 2010, 29: 693-699.View ArticleGoogle Scholar
- Dedon PC: Determination of binding mode: intercalation. Curr Protoc Nucleic Acid Chem. 2001, 8: 1-13.PubMedGoogle Scholar
- Kanakis CD, Tarantilis PD, Pappas C, Bariyanga J, Tajmir-Riahi HA, Polissiou MG: An overview of structural features of DNA and RNA complexes with saffron compounds: models and antioxidant activity. J Photochem Photobiol. 2009, 95: 204-212.View ArticleGoogle Scholar
- Clarke EGC, Clarke ML: Veterinary Toxicology. Edited by: Cassel , Collier . 1997, London: Macmillan publishers, 268-277.
- Wakte PS, Sachin BS, Patil AA, Shinde DB: Hepatoprotective activity of Acacia nilotica flowers. Med Chem Drug Discovery. 2012, 3: 152-159.Google Scholar
- Guta M, Urga K, Assefa A, Lemma H, Addis G, Gemeda N, Yirsaw k, Mudi K, Melaku D: Antibacterial and acute toxicity study of Acacia nilotica. Ethiop J Biol Sci. 2007, 6: 43-49.Google Scholar
- El-Hadiyah TM, Abdulhadi NH, Badico EEM, Mohammed EYG: Toxic potential of ethanolic extract of Acacia nilotica (Garad) in rats. Sudan J Med Sci. 2011, 6: 1-6.Google Scholar
- Raza M, Al-Shabanah OA, El-Hadiyah TM, Al-Majed AA: Effect of prolonged vigabatrin treatment on haematological and biochemical parameters in plasma, liver and kidney of Swiss albino mice. Sci Pharm. 2002, 70: 135-145.Google Scholar
- Teo S, Stirling D, Thomas S, Hoberman A, Kiorpes A, Khetani V: A 90-day oral gavage toxicity study of d-methylphenidate and d I-methylphenidate in Sprague-Dawley rats. Toxicol. 2002, 179: 183-196.View ArticleGoogle Scholar
- Al-Mustafa ZH, Dafallah AA: A study on the toxicology of Acacia nilotica. Am J Chin Med. 2000, 28: 123-View ArticlePubMedGoogle Scholar
- Rodostits OM, Gay CC, Blood DC, Hinchcliff KW: Clinica Veterinaria. 2002, Rio de Janeiro, RJ, Brazil: Guanabara Koogan, Rio de Janeiro, 1737-9Google Scholar
- Wolford ST, Schroer RA, Gohs FX, Gallo PP, Brodeck M, Falk HB, Ruhren R: Reference range data base for serum chemistry and hematology values in laboratory animals. J Toxicol Environ Health. 1986, 18: 161-188.View ArticlePubMedGoogle Scholar
- Johnson-Delaney C: Small mammals. Exotic animal companion medicine handbook for veterinarians. Edited by: Hudelson KS. 1996, Florida: Zoological Education Network, 1-62.Google Scholar
- Asad M, Munir TA, Afzal N: Acacia nilotica leaf extract and glyburide: comparison of fasting blood glucose, serum insulin, beta-thromboglobulin levels and platelet aggregation in streptozotocin induced diabetic rats. J Pak Med Assoc. 2011, 61: 247-251.PubMedGoogle Scholar
- Corns CM: Herbal remedies and clinical biochemistry. Annals of Clin Biochem. 2003, 40: 489-507.View ArticleGoogle Scholar
- Hilaly J, El Israili ZH, Lyoussi B: Acute and chronic toxicological studies of Ajuga iva in experimental animals. J Ethnopharmacol. 2004, 91: 43-50.View ArticlePubMedGoogle Scholar
- Isnard BC, Deray G, Baumelou M, Le Quintree M, Vanherweghem JL: Herbs and the kidney. Am J Kidney Disease. 2004, 44: 1-11.View ArticleGoogle Scholar
- Saad B, Azaizeh H, Abu-Hijleh G, Said S: Safety of traditional Arab herbal medicine. Evid Based Complement Alternat Med. 2006, 3: 433-439.View ArticlePubMedPubMed CentralGoogle Scholar
- Tolman KG, Rej R: Liver function. Tietz Textbook of clinical chemistry. Edited by: Burtis CA, Ashwood ER. 1999, Philadelphia Pennsylvania: Saunders Company, 1125-1177. 3Google Scholar
- Mitruka BM, Rawnsley HM: Clinical, Biochemical and Hematology reference values in normal and experimental animals. Clinical, biochemical and hematological reference values in normal experimental animals. Edited by: Mitruka BM, Rawnsley HM. 1981, USA: Masson publishing, 134-135. 2Google Scholar
- Kannan N, Sakthivel KM, Guruvayoorappan C: Protective effect of Acacia nilotica (L.) against acetaminophen-induced hepatocellular damage in wistar rats. Adv Pharmacol Sci. 2013, 2013: 1-9.View ArticleGoogle Scholar
- The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1472-6882/14/257/prepub
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