Total phenolic, flavonoid contents, in-vitro antioxidant activities and hepatoprotective effect of aqueous leaf extract of Atalantia ceylanica

BMC Complementary and Alternative Medicine

Preethi Soysa¹  Chamira Dilanka Fernando¹ 

[1] Department of Biochemistry & Molecular Biology, Faculty of Medicine, University of Colombo, Kynsey Road, Colombo 08, Sri Lanka

关键词: Hepatoprotective;    Antioxidants;    Aqueous extract;    Atalantia ceylanica;    Flavonoids;    Phenolics;   

Others  :  1086050
DOI  :  10.1186/1472-6882-14-395

 received in 2014-05-15, accepted in 2014-10-06,  发布年份 2014

【 摘 要 】

Background

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. Lyophilized powder of the water extract of A. ceylanica leaves was investigated for its phytochemical constituents, antioxidant and hepatoprotective activity in-vitro.

Methods

The 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.

Results

The 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).

Conclusions

The 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.

【 授权许可】

   
2014 Fernando and Soysa; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

【 参考文献 】

• [1]McCord JM: The evolution of free radicals and oxidative stress. Am J medi Sci 2000, 108:652-659.
• [2]Hikino H, Kiso Y: Natural Products for Liver Disease. London: Academic Press; 1988:39-42.
• [3]Liao KL, Yin MC: Individual and combined antioxidant effects of seven phenolic agents in human erythrocyte membrane ghosts and phosphatidyl choline liposome systems: Importance of the partition coefficient. J Agric Food Chem 2000, 48:2266-2270.
• [4]Ahmadi A, Ebrahimzadeh MA, Ahmad-Ashrafi S, Karami M, Mahdavi MR, Saravi SSS: Hepatoprotective, antinociceptive and antioxidant activities of cimetidine, ranitidine and famotidine as histamine H2 receptor antagonists. Fundam Clin Pharmacol 2011, 25(1):72-79.
• [5]Jayaprakash GK, Rao LJ: Phenolic constituents from lichen Parmontrema stuppeum”. Food Control 2000, 56:1018-1022.
• [6]Momin A: Role of Indigenous Medicine in Primary Health Care. In Proceedings of the first International Seminar on Unani Medicine. New Delhi, India; 1987:54.
• [7]Dassanayake MD, Forsberg FR: A Revised Handbook to the Flora of Ceylon, Volume 5. New Delhi:Oxford: IBH Publishing Co; 1985:406-475.
• [8]Jayaweera DMA: Medicinal Plants (Indigenous and Exotic) Used in Ceylon, Part V. Colombo: A publication of the National Science Council of Sri Lanka; 1982.
• [9]Makkar HPS, Blummel M, Borowy NK, Becker K: Gravimetric determination of tannins and their correlations with chemical and protein precipitation methods. J Sci Food Agric 1993, 61:161-165.
• [10]Zhishen J, Mengcheng T, Jianming W: The determination of flavonoid contents in mulberry and their scavenging effects on Superoxide radicals. Food Chem 1999, 64:555-559.
• [11]Silva IK, Soysa P: Evaluation of phytochemical composition and antioxidant capacity of a decoction containing adenanthera pavonina L. and Thespesia populnea L. Pharmacogn Mag 2011, 7(27):193-199.
• [12]Halliwell B, Gutteridge JM, Arnoma OL: The Deoxyribose method: A simple test tube assay for the determination of rate constant for reaction of hydroxyl radical. Anal Biochem 1987, 165:215.
• [13]Garret D: The quantitative analysis of drugs. Champman Hall Japan 1964, 3:456-458.
• [14]Oyaizu M: Studies on products of browning reactions: antioxidative activities of products of browning reaction prepared from glucosamine. Jpn J Nutr 1986, 44:307-315.
• [15]Wormser U, Ben ZS, Stivelband E, Eizen O, Nyska A: The Liver slice system: A rapid in vitro acute toxicity test for primary screening of hepatotoxic agents. Toxicol in Vitro 1990, 4:783-789.
• [16]Invittox protocol no.42: The ERGATT/FRAME Data Bank of in Vitro Techniques in Toxicology: Liver Slice Hepatotoxicity Screening System. England: INVITTOX; 1992.
• [17]Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurement with the Folin phenol reagent. J Biol Chem 1951, 193:265.
• [18]Lynch SM, Frei B: Mechanisms of copper- and iron dependent oxidative modification of human low-density lipoprotein. J Lipid Res 1993, 34:1745-1751.
• [19]Dhalwal K, Shinde VM, Namdeo AG: Antioxidant profile and HPTLC densitometric analysis of umbelliferone and psoralen in Aegle marmelos. Pharma Biol 2008, 46:266-272.
• [20]Reddy KH, Sharma PV, Reddy OV: A comparative in vitro study on antifungal and antioxidant activities of Nervilia aragoana and Atlantia monophylla. Pharm Biol 2010, 48(5):595-602.
• [21]Pawar CR, Surana SJ: Optimizing conditions for gallic acid extraction from Caesalpinia decapetala wood. Pak J Pharm Sci 2010, 23(4):423-425.
• [22]Oboh G, Puntel RL, Rocha JBT: Hot pepper (Capsicum annuum, Tepin and Capsicum chinese, Habanero) prevents Fe2+-induced lipid peroxidation in brain in vitro. Food Chem 2007, 102:178-185.
• [23]Siddique NA, Mujeeb M, Najmi AK, Akram M: Evaluation of antioxidant activity, quantitative estimation of phenols and flavonoids in different parts of Aegle marmelos. African J Plant Sci 2010, 4(1):001-005.
• [24]Molyneux P: The use of the stable free radical diphenylpicrylhydrazyl (DPPH) for estimating antioxidant activity Songklanakarin. J Sci Technol 2004, 26(2):211-219.
• [25]Mishra J, Yousuf A, Singh RD, Aradhana : Phytochemical investigation and in-vitro antioxidant potential of leaves of Murraya koenigii. Int J Integr Biol 2009, 7(3):171.
• [26]Pandey P, Mehta A, Hajra S, John J, Mehta P: Antioxidant property, total Phenolic content and inhibition of a-amylase activity of Ruta graveolens L. leaves extract. J Pharm Res 2011, 4(6):1735-1737.
• [27]Muthiah PL, Umamaheswari M, Asokkumar K: In-vitro antioxidant activities of leaves, fruits and peel extracts of Citrus. Int J Phytopharmacy 2012, 2(1):13-20.
• [28]Malinski T: Nitric oxide and nitroxidative stress in Alzheimer's disease. J Alzheimers Dis 2007, 11:207-218.
• [29]Attarde DL, Chaudhari BJ, Bhambar RS: Phytochemical investigation and in vitro antioxidant activity of extracts from leaves of Limonia acidissima linn. (Rutaceae). J Pharm Res 2011, 4(3):766-768.
• [30]Yen GC, Chen HY: Antioxidant activity of various tea extracts in relation to their antimutagenicity. J Agric Food Chem 1995, 43:27-32.
• [31]Naik RS, Mujumdar AM, Ghaskadbi S: Protection of liver cells from ethanol cytotoxicity by curcumin in liver slice culture in vitro. J Ethnopharmacol 2004, 95:31-37.
• [32]Mahnaaz M, Swati K, Devasagayam TP, Saroj G: Pterocarpus marsupium extract reveals strong in vitro antioxidant activity. Drug Discov Ther 2009, 3:151-161.
• [33]Grosse-Siestrup C, Pfeffer J, Unger V, Nagel S, Witt C, Fischer A, Groneberg DA: Isolated hemoperfused slaughterhouse livers as a valid model to study hepatotoxicity. Toxicol Pathol 2002, 30:749.
• [34]Vilei MT, Granato A, Ferraresso C, Neri D, Carraro P, Gerunda G, Muraca M: Comparison of pig, human and rat hepatocytes as a source of liver specific metabolic functions in culture systems–implications for use in bioartificial liver devices. Int J Artif Organs 2001, 24(6):392-396.
• [35]Drotman RB, Lawhorn GT: Serum enzymes as indicators of chemical induced liver damage. Drug Chem Toxicol 1978, 1:163-171.
• [36]Christina AJ, Saraswathy GR, Robert SJ, Kothai R, Chidambaranathan N, Nalini G, Therasal RL: Inhibition of CCl4-induced liver fibrosis by Piper longum Linn. Phytomedicine 2006, 13:196-198.
• [37]Pal R, Nath R, Gill KD: Lipid peroxidation and antioxidant defense enzymes in various regions of adult rat brain after coexposure to cadmium and ethanol. Pharmacol Toxicol 1993, 73:209-214.
• [38]Habibi E, Shokrzadeh M, Chabra A, Naghshvar F, Keshavarz Maleki R, Ahmadi A: Protective effects of Origanum vulgare ethanol extract against cyclophosphamide-induced liver toxicity in mice. Pharm Biol 2014, 15:1-6.
• [39]Sinha S, Bhat J, Joshi M, Sinkar V, Ghaskadbi S: Hepatoprotective activity of Picrorhiza kurroa Royle Ex. Benth extract against alcohol cytotoxicity in mouse liver slice culture. Int J Green Pharmacy 2011, 5(3):244-253.
• [40]Shokrzadeh M, Chabra A, Ahmadi A, Naghshvar F, Habibi E, Salehi F, Assadpour S: Hepatoprotective effects of zataria multiflora ethanolic extract on liver toxicity induced by Cyclophosphamide in mice. Drug Res 2014, 64:1-7.