Hepatoprotective effects of partially purified fractions of Senna occidentalis ethanolic extract on diethyl nitrosamine-induced toxicity in Wistar rat

Ojochenemi E. Yakubu, Eleojo B. Ojogbane, Francis O. Atanu, Chukwuka S. M. Udeh, Morayo E. Ale, Blessing H. Bello


Background: Induction of toxicity using nitrosamines provides a reliable animal model for the study of oxidative damage to lipids, cellular membranes, proteins and DNA. In the present report, the effects of partially purified fractions of Senna occidentalis leaves on diethylnitrosamine intoxicated rats were studied.

Methods: Fractions obtained from eluting the column with solvents of increasing polarity, n-hexane, chloroform, ethyl acetate, ethanol, methanol and distilled water were subjected to in vitro for their ability to scavenge 1, 1-dipheny l, 2-pycryl hydrazyl (DPPH) radical. Fraction 6a eluted with ethyl acetate:ethanol (50:50) possessed the highest antioxidant activity, this fraction was therefore selected for in vivo studies. Twenty rats, each weighing between 150 to 250 g were randomly allocated into four groups of five rats each. Hepatotoxicity was induced using a single intraperitoneal injection of diethylnitrosamine (DEN) at the 200 mg/kg body weight. Treatment was carried out for 3 weeks by oral gavage as follows: group A, normal control, group B, DEN control, group C, DEN+fraction (10 mg/kg), group D, DEN+silymarin (5 mg/kg).

Results: The results showed that DEN toxicity significantly (p<0.05) increased alanine transaminase (ALT) and aspartate transaminase (AST) activities and increased the level of thiobarbituric acid reactive substance (TBARS) in the liver. In contrast, the levels of bilirubin, total protein (TP) and albumin (ALB) decreased. However, treatment of rats with the extract significantly (p<0.05) reduced the concentrations of TBARS, ALT, AST and bilirubin, but increased the concentration of TP and ALB.

Conclusions: These results show hepatoprotective potentials of the fraction. Furthermore, GC-MS fingerprinting of fraction 6a revealed the presence of compounds with anticancer, antioxidant and anti-inflammatory properties confirming its high chance for exploration as a medicinal agent.


Hepatoprotective, Senna occidentalis, Diethylnitrosamine, Toxicity, Antioxidant

Full Text:



Gupta M, Mazumder UK, Kumar RS, Kumar T. Antitumour activity and antioxidant role of Bauhinia racemosa against Ehrlisch ascites carcinoma in swiss albino mice. Acta Pharmacol Sin. 2004;25(8):1070-6.

Tolba H, Moghrani H, Benelmouffok A, Kellou D, Maachi R. Essential oil of Algerian Eucalyptus citriodora: Chemical composition, antifungal activity. Journal de Mycologie M├ędicale. 2015;25(4):128-33.

Sylla BS, Wild CP. A million Africans a year dying from cancer by 2030: what can cancer research and control offer to the continent. Int J Cancer. 2012;130(2): 245-50.

Veronique JE, Gabriel NM. Sub-chronic toxicity of the beverage made from Cassia occidentalis seeds in mice. Int J Nutrit Food Sci. 2013;2(5):237-42.

Yakubu OE, Nwodo OFC, Joshua PE, Ugwu MN, Odu AD, Okwo F. Fractionation and determination of total antioxidant capacity, total phenolics and total flavonoids contents of aqueous, ethanol and n-Hexane extracts of Vitex doniana leaves. Afr J Biotechnol. 2014;13(5):693-8.

Karadag A, Ozcelik B, Saner S. Review of methods to determine antioxidant capacities. Food Analyt Method. 2009;2:41-60.

Ortuno J, Serrano R, Jordan MJ, Banon S. Relationship between antioxidant status and oxidative stability in lamb meat reinforced with dietary rosemary diterpenes. Food Chem. 2016;190:1056-63.

Reitman S, Frankel S. A colorimetric method for the determination of serum glutamic oxalacetic and glutamic pyruvic transaminases. Am J Clinic Pathol. 1957;28(1):56-63.

Doumas BT, Watson WA, Biggs HG. Albumin standards and the measurement of serum albumin with bromocresol green. Clinica Chimica Acta. 1997;258(1):87-96.

Bartholomew RJ, Delaney AM. Sulphonphthaleins as specific reagents for albumin: determination of albumin in serum. Proc Aust Assoc Clin Biochemists. 1966;1:214-8.

Fine J. Quantitative determination of serum proteins by colorimetric method. Biochem J. 1935;29:799.

Koleva II, Beek TAV, Linssen JPH, DeGroot A, Evstatieva LN. Screening of plant extracts for antioxidant activity: a comparative study on three testing methods. Phytochem Analysis. 2002;13(1):8-17.

Engoor SA, Mangalanandan SL, Paul HC, Jagathala MS. In vitro antioxidant, anti-lipid peroxidation activities and HPLC analysis of methanol extracts from bark and stem of Mahonia leschenaultia takeda. Asian J Plant Sci Res. 2013;3(2):116-26.

Fatma A, Sokindra K, Shah AK. Estimation of total phenolic content, in-vitro antioxidant and anti-inflammatory activity of flowers of Moringa oleifera. Asian Pac J Trop Biomed. 2013;3(8):623-7

Vivek KB, Yong-Ha P, Pooja, A. Studies on phytochemical analysis, antioxidant and lipid peroxidation inhibitory effects of a medicinal plant, Coleusforskohlii. Frontiers Life Sci. 2015;8:2:139-47.

Richa U, Jitendra KC, Kavindra NT, Karuna S. Antioxidant property of aerial parts and root of Phyllanthus fraternus webster, an important medicinal plant. Scient World J. 2014:692392.

Coppen PP. In: Allen JC, Hamilton RJ, eds. Rancidity in foods. New York, USA: Applied Science Publishers; 1983: 67.

Tuyen TKN, Chamroon L, Montinee T, Komkhae P. Potential antioxidant and lipid peroxidation inhibition of Phyllanthus acidus leaf extract in minced pork. Asian Australas J Animal Sci. 2017;30(9):1323-31.

Balu M, Sangeetha P, Haripriya D, Panneerselvam C. Rejuvenation of antioxidant system in central nervous system of aged rats by grape seed extract. Neurosci Letter. 2005;383:295-300.

Patrick-Iwuanyanwu KC, Wegwu MO, Okiyi JK. Hepatoprotective effects of African locust bean (Xylopia aethiopica) in CCl4 induced liver damaged Wistar albino rats. Int J Pharmacol. 2010;6:744-9.

Hites AR. Gas Chromatography Mass Spectroscopy: Handbook of Instrumental Techniques for Analytical Chemistry. Upper Saddle River, NJ: Prentice Hall PTR; 1997: 609-11.