Elsevier

Toxicology in Vitro

Volume 23, Issue 6, September 2009, Pages 1034-1038
Toxicology in Vitro

Leaves of Cassia tora as a novel cancer therapeutic – An in vitro study

https://doi.org/10.1016/j.tiv.2009.06.010Get rights and content

Abstract

Cassia tora Linn (Leguminacea) is a medicinal plant traditionally used as laxative, for the treatment of leprosy and various skin disorders. Preliminary phytochemical analysis of leaf showed the presence of polyphenols (3.7 mg gallic acid equivalent per gram dried leaves). The presence of phenolic compound prompted us to evaluate its antioxidant and antiproliferative potential. In the present study C. tora methanolic leaf extract (CTME) was evaluated for its nitric oxide scavenging activity and reducing power assays using Rutin and BHT as standards. The extract was studied for its lipid peroxidation inhibition assay using rat liver and brain. In all assays, a correlation existed between concentration of extract and percentage inhibition of free radical, reducing power and inhibition of lipid peroxidation. The antiproliferative activity of CTME with Cisplatin, anticancer drug was studied using human cervical cancer cells (HeLa). Proliferation of HeLa was measured by MTT assay, cell DNA content by modified diphenylamine method and apoptosis by Caspase 3 activity. The plant extract induced a marked concentration dependent inhibition on proliferation, reduced DNA content and apoptosis in HeLa. These results clearly indicate that C. tora is effective against free radical mediated diseases.

Introduction

The ancient Indian system of medicine relies predominantly on plant based drugs. Majority of the diseases/disorders are mainly linked to oxidative stress due to free radicals (Gutteridge, 1993). Antioxidants act as a major defense against radical mediated toxicity by protecting the damages caused by free radicals (Lee et al., 2003). They can act as radical scavengers, inhibiting lipid peroxidation and other free radical mediated processes protecting the human organs against several pathologies attributed to radical reactions such as Parkinson’s disease, atherosclerosis, Alzheimer’s disease and cancer (Emerit et al., 2004, Mates et al., 1999, Rosenkranz, 2002). Among the natural antioxidants, polyphenols which are distributed in the plant kingdom reported to have beneficial health effects including anti-inflammatory, anticarcinogenic and vasodilatory action (Middleton et al., 2000, Hotta et al., 2002). These protective effects have been mostly ascribed to their scavenging, metal chelating and antioxidant action (Kameoka et al., 1999). Thus, phenolic compounds possess significant antioxidant capacity that may be associated with lower incidence and lower mortality rates of cancer in several human populations (Velioglu et al., 1998).

Apoptosis is characterized by a series of morphological changes involving cell shrinkage, chromatin condensation and the formation of apoptotic bodies (Borner, 2003). Apoptosis can be triggered by various extracellular and intracellular stimuli that result in the coordinated activation of a family of proteases called caspases. The activation of caspases plays a pivotal role in the execution of cell apoptosis (Polverino and Patterson, 1997). The Caspase 3 activation pathway is an important downstream executioner in apoptosis.

Cassia tora (CT) (family:leguminacea) is a common edible leafy vegetable consumed by Indians. It is a well known ayurvedic medicinal plant as a laxative, antiperiodic and is useful for leprosy, ringworm, bronchitis and cardiac disorders. It was reported that seeds of CT has antioxidant activity and contain many active substances including chrysophenol, emodin, rhein etc. (Huang, 1993). Scientific evaluation of CT leaves on antioxidant and antiproliferative effect is not yet carried out. In India, majority of the population use traditional natural preparations of plant leaf for the treatment of various diseases and also as diet. So the objectives of this study was to assess the antioxidant and antiproliferative potential of this leaf extract.

Section snippets

Chemicals

3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide (MTT), fetal calf serum (FCS), Griess reagent, Caspase 3 assay kit and dimethyl sulfoxide (DMSO) were purchased from Sigma (St. Louis, USA). All other reagents used were of analytical grade.

Plant materials

CT was collected from different parts of Thiruvananthapuram, authenticated by an expert and deposited in the herbarium of the Department of Botany, University of Kerala, Thiruvananthapuram, India. The fresh leaves of CT was washed thoroughly and

Phenolic content of CTME

The total phenolic content of CT was found to be 3.7 mg gallic acid equivalent per gram dried leaves.

Nitric oxide scavenging activity

The antioxidant activity of CTME was on the basis of their ability to scavenge NO radical. IC50 were estimated as 140 μg/ml for Rutin and for CTME it was 180 μg/ml and the result was shown in Fig. 1.

Reducing power

The reducing power of CTME was shown in Fig. 2 with BHT as positive control. CTME exhibited a dose dependent reducing power activity.

Lipid peroxidation inhibition

Lipid peroxidation inhibition by CTME was concentration dependent. IC

Discussion

CT is one of the widely used drugs in various ayurvedic and herbal formulations. The phytochemical analysis of CTME showed significant amount of total phenol. Numerous polyphenolic substances isolated from medicinal plants were found to exhibit antioxidant, antitumor and antiviral properties depending on the chemical structure of each polyphenol (Okuda et al., 1992, Yoshida et al., 1992). It was suggested that polyphenolic compounds have inhibitory effects on mutagenesis and carcinogenesis in

Acknowledgements

The authors are grateful to Dr. Priya Srinivas, Scientist E1, Molecular Therapeutics Laboratory, Rajiv Gandhi Centre for Biotechnology, Trivandrum, India for providing the cell lines for the study. Also thanks are due to Dr. Valsala Devi, Curator, Department of Botany, University of Kerala, Thiruvananthapuram for authentication of the plant material.

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