Pak. J. Pharm. Sci., Vol.27, No.6(Suppl), November 2014, pp.2101-2107 2101 Phytochemical and therapeutic evaluation of leaf and in vitro derived callus and shoot of Solanum trilobatum Priya Govindarajan 1 and Chellaram Chinnachamy 2* 1 Department of Biotechnology, Sathyabama University, Chennai, Tamilnadu, India 2 Department of Biomedical Engineering, Vel Tech Multitech Engineering College, Chennai, Tamilnadu, India Abstract: This study focuses on the phytochemical properties and the anti-hepatocarcinogenic effects of the leaf and in vitro-derived callus and shoot extracts of Solanum trilobatum. In the leaf, callus and shoot, the presence of sugar, proteins, alkaloids, flavonoids, saponins, tananins, cardiacglycoside, terpenoid and lipids was established by preliminary phytochemical screening. Surface-sterilized explants (0.5-1.0 cm) were placed on the MS basal medium supplemented with different concentrations of 2,4-dichlorophenoxyacetic acid (0.45, 2.26, 4.52, 11.31 and 12.56µM), naphthylacetic acid (NAA; 0.54, 1.34, 2.69, 5.37, 13.43 and 26.85µM) and 6-benzylaminopurine (BA; 0.44, 1.11, 2.22, 4.44, 8.88 and 13.32µM) for callus induction. Explants from node and callus culture were inoculated on the MS basal medium supplemented with varying concentrations of BA (0.44-22.20 µM) and NAA (0.54-10.74µM) for shoot multiplication. Rats were divided into five groups and administered with diethyl nitrosamine (DEN) and DEN (200mg/kg bwt) intraperitoneally along with methanol leaf and in vitro-derived callus and shoot extracts (250mg/kg bwt) orally for 3 months. A significant deviation (P<0.05) in marker enzymes such as alanine transaminase, aspartate transaminase, lactate dehydrogenase and total bilirubin was found in rats administered with DEN. The liver tissue was used for the analysis of glutathione reductase, lipid peroxidation, glutathione peroxidase, glutathione S-transferase, superoxide dismutase and catalase. DEN administration caused a significant elevation in serum enzymes and total bilirubin. Moreover, antioxidant enzymes were drastically inhibited with significant reduction in glutathione and increased lipid per oxidation. Increased glutathione level and reduced lipid peroxidation were also evident in S. trilobatum-treated rats. However, crude S. trilobatum and in vitro-derived callus and shoot extracts offered better protection against free radical toxicity induced by DEN. Keywords: Diethyl nitrosamine, free radical, antioxidant, liver, Solanum trilobatum. INTRODUCTION Recently, there is a renewed interest worldwide in plants as pharmaceuticals. The discovery of numerous biologically active molecules by the pharmaceutical industry and the increasing use of crude extracts of plants for self-medication by the general public are clear evidences (Chellaram et al., 2009a, 2011). Several of these herbal drugs are extracted from plants growing in wild strands. Nearly 95% of plants used as ingredients in traditional drugs are collected from forests and other natural reserves (Nadkarani, 1979). However, limited distribution, low availability, inaccessibility and anthropogenic pressure have caused a large decline in several of the wild populations, making them rare species. Solanum trilobatum Linn (family: Solanaceae) has been one of the common Indian medicinal plants used for many centuries to treat asthma, cough, dyspnoea, chronic febrile infections and parturition. It shows cardiac, tonic and carminative actions. Sobatum, β solamarine, solaine, solasodine, glycoalkaloid, disgenin and tomatidine are its chief constituents. This plant has been reported to possess a broad spectrum of antibiotic, antibacterial and anticancer activities (Mohanan, 1996). The active agent (sobatum) obtained by petroleum ether extract was demonstrated to have anticancer properties by in vitro and in vivo experiments. The existing variability in a breeding population may not be sufficient for modern plant- breeding purposes; thus an effort has to be made to broaden the existing gene pool of crops. Because of these complications and adverse ecological impacts plant cell culture have been used as an alternative source of the natural products and novel lead compounds with medicinal properties (Chellaram et al., 2009b and Prem Anand, 2011). Primary liver cancer ranks fifth in frequency among all malignancies in the world. A vast majority of primary liver cancer is hepatocellular carcinoma (HCC). HCC frequently occurs in the form of a solid tumor. It is highly prevalent with 620,000 new cases reported per year worldwide, of which more than 80% of cases are being reported from China, Africa and South East Asia (Ribes et al., 2008). It is highly aggressive, with around 595,000 deaths per year (Parkin et al., 2000). HCC presents with limited therapeutic options. Therefore, more research focusing on the biological bases of this malignancy is anticipated in order to develop new strategies for treatment. An animal model of diethyl nitrosamine *Corresponding author: e-mail: chellarampublications@gmail.com