Preliminary evaluation of in vitro cytotoxicity and in vivo antitumor activity of Premna herbacea Roxb. in Ehrlich ascites carcinoma model and Dalton\'s lymphoma ascites model

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Author's personal copy Experimental and Toxicologic Pathology 65 (2013) 235–242

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Preliminary evaluation of in vitro cytotoxicity and in vivo antitumor activity of Premna herbacea Roxb. in Ehrlich ascites carcinoma model and Dalton’s lymphoma ascites model Isha Dhamija, Nitesh Kumar, S.N. Manjula, Vipan Parihar, M. Manjunath Setty, K.S.R. Pai ∗ Manipal College of Pharmaceutical Sciences, Manipal University, Manipal-576104, India

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Article history: Received 4 May 2011 Accepted 18 August 2011 Keywords: Brine shrimp lethality assay Trypan blue dye exclusion assay MTT assay Antioxidant

a b s t r a c t In the present study, the root nodules of Premna herbacea Roxb. (PH) was investigated for its in vitro cytotoxicity and in vivo antitumor activity. Two extracts, aqueous and alcoholic; two fractions of alcoholic extract, ethyl acetate and butanol fractions were screened for their in vitro cytotoxicity by brine shrimp lethality (BSL) assay, trypan blue exclusion assay and MTT assay. Alcoholic extract and its ethyl acetate fraction were found to be the most effective in BSL assay, trypan blue exclusion assay. In vivo antitumor activity was screened in the Ehrlich ascites carcinoma (EAC) model and the Dalton lymphoma ascites (DLA) model. The extracts and the fractions were tested at two dosages (250 and 500 mg/kg) by intraperitoneally (i.p.) route on every alternate day upto 13th day. Cisplatin was used as positive control in both studies in single dose (day 1) 3.5 mg/kg by i.p. route. In EAC model, ascites tumor was induced by inoculating 2.5 million of EAC cells i.p. alcoholic extract at 500 mg/kg was the most effective in elevating MST, reduction in body weight in EAC induced tumor. Only the effective extract i.e., alcoholic extract were studied for hematological and antioxidant parameter. It showed a restoring effect on altered hematological parameters and a significant improvement in biochemical parameters at 250 mg/kg dose of alcoholic extract. These results explain the toxicity of 500 mg/kg might be high. In the Dalton lymphoma ascites (DLA) model, solid tumor was developed by i.m. injection of 1 million DLA cells. Both the extracts and the fractions possessed potent antitumor activity against solid tumor models by significantly reducing the solid tumor weight and volume. © 2011 Elsevier GmbH. All rights reserved.

1. Introduction Cancer therapy has been remarkably consistent with the last 50 years. Surgery, radiation, and chemotherapy have been the cornerstones of conventional treatment. Not surprisingly, the clinical success of these treatments has reached a plateau (Braverman, 1991). Today, combating cancer has paramount importance and numerous multidisciplinary scientific approaches have been applied to treat this disease but the sure-shot, perfect cure is yet to be brought into the world of medicine. The present western system of medicine is embodied with severe unbearable complications and patient incompliance. Hence, a more specific and safe remedy is the need of the hour. Recent studies have shown that cancer management by phytochemicals obtained from vegetables, fruits, spices, teas, herbs and medicinal plants, is one of the most feasible approaches to treat this cascading disease. Plants have been a

∗ Corresponding author at: Department of Pharmacology, Manipal College of Pharmaceutical Sciences, Manipal University, Manipal-576104, Karnataka, India. Tel.: +91 820 2922482; fax: +91 820 2571998. E-mail address: [email protected] (K.S.R. Pai). 0940-2993/$ – see front matter © 2011 Elsevier GmbH. All rights reserved. doi:10.1016/j.etp.2011.08.009

source of medicinal substances for thousands of years, and phytoproducts continue to play an essential role in medicine. Plants have a long history of use in the treatment of cancer and are still in use (Huang, 1999). They have been the source of some of the currently available anticancer agents such as vinblastine, vincristine, etoposide, teniposide, and paclitaxel (Cragg et al., 1997). Among them the best known are so called vinca alkaloids (vinblastine and vincristine) isolated from the Madagascar periwinkle, Catharanthus roseus (Vinca rosea L., Apocinaceae). C. roseus was used by various cultures for the treatment of diabetes while vinblastine and vincristine were first discovered during an investigation of the plant as a source of potential oral hypoglycemic agents (Cragg et al., 1997). The rich and diverse plant sources in India are likely to provide effective anticancer agents. One of the best approaches in the search for anticancer agents from plant resources is the selection of plants based on ethno-medical leads. Here, the plant, Premna herbacea Roxb. (PH) known as Sirutekku in Tamil is used in Siddha, the traditional system of medicine practised in south India. It is claimed to be useful in the ayurvedic system of medicine either alone or as an ingredient in compound preparations for the treatment of several ailments, such as bronchitis, asthma, blood pressure, tumors, inflammation, cough, epilepsy,

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