Abstract
BackgroundDecoction prepared from leaves of Atalantia ceylanica is used in traditional medicine in Sri Lanka for the treatment of various liver ailments since ancient times. Lyophilized powder of the water extract of A. ceylanica leaves was investigated for its phytochemical constituents, antioxidant and hepatoprotective activity in-vitro.MethodsThe total phenolic and flavonoid contents were determined using Folin Ciocalteu method and aluminium chloride colorimetric assay respectively. The antioxidant activities of the decoction were investigated using 1,1-Diphenyl-2-picrylhydrazyl (DPPH), hydroxyl radical, nitric oxide scavenging assays and ferric ion reducing power assay. Hepatotoxicity was induced on porcine liver slices with ethanol to study hepatoprotective activity. Porcine liver slices were incubated at 37°C with different concentrations of the water extract of A. ceylanica in the presence of ethanol for 2 hours. The hepatoprotective effects were quantified by the leakage of alanine transaminase (ALT), aspartate transaminase (AST) and lactate dehydrogenase (LDH) to the medium. Thiobarbituric acid reactive substances (TBARS) assay was performed to examine the anti-lipid peroxidation activity caused by the plant extract.ResultsThe mean ± SD (n =9) for the levels of total phenolics and flavonoids were 4.87 ± 0.89 w/w% of gallic acid equivalents and 16.48 ± 0.63 w/w% of (-)-Epigallocatechin gallate equivalents respectively. The decoction demonstrated high antioxidant activity. The mean ± SD values of EC50 were 131.2 ± 36.1, 48.4 ± 12.1, 263.5 ± 28.3 and 87.70 ± 6.06 μg/ml for DPPH, hydroxyl radical, nitric oxide scavenging assays and ferric ion reducing power assay respectively.A significant decrease (p <0.05) was observed in ALT, AST and LDH release from porcine liver slices treated with A. ceylanica extract at a concentration of 2 mg/ml in the presence of ethanol (5 M) compared to that of ethanol (5 M) treated slices. Furthermore, a reduction in lipid peroxidation was also observed in liver slices treated with the leaf extract of A. ceylanica (2 mg/ml) compared to that of ethanol induced liver toxicity (p <0.05).ConclusionsThe results suggest that aqueous extract of A. ceylanica exerts hepatoprotective activity against ethanol induced liver toxicity of porcine liver slices which can be attributed to the antioxidant properties possessed by the plant material.Electronic supplementary materialThe online version of this article (doi:10.1186/1472-6882-14-395) contains supplementary material, which is available to authorized users.
Highlights
Decoction prepared from leaves of Atalantia ceylanica is used in traditional medicine in Sri Lanka for the treatment of various liver ailments since ancient times
Phenolic compounds contain hydroxyl groups which contribute to the free radical scavenging and act as primary antioxidants [19]
A similar study carried out using the ethanolic leaf extract of Atalantia monophylla (Family: Rutaceae), has shown that the total phenolic content present was 560 mg/g (56% w/w% gallic acid equivalents) which is very high compared to the present study [20]
Summary
Chemicals and equipment The chemicals gallic acid, Folin Ciocalteu reagent, trichloroacetic acid, 2-deoxy-D-ribose and ethylenediamine tetra acetic acid (EDTA) were purchased from Sigma Chemicals Co. The effective concentration needed to scavenge DPPH free radical by 50% (EC50) was calculated by regression analysis of the dose response curve plotted between percentage inhibition versus concentration of the test samples and the standard. The percentage scavenging of hydroxyl radical for A. ceylanica and the standard antioxidant was calculated according to equation 1. Dose response curve was plotted between the absorbance versus concentrations of plant extracts or standards by linear regression analysis. The supernatants of the tissue homogenates and the media obtained from post incubation of tissues were assayed for alanine transaminase (ALT), aspartate transaminase (AST) and lactate dehydrogenase (LDH).The percentage cytotoxicity was calculated using equation 2: The percentage cytotoxicity 1⁄4 Enzyme activity in the medium  100 Total Enzyme activity ð2Þ. EC50 values were calculated from either linear or logarithmic dose response curves where R2 > 0.90
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