Protection against diethylnitrosoamine-induced hepatocarcinogenesis by an indigenous medicine comprised of Nigella sativa, Hemidesmus indicus and Smilax glabra : a preliminary study

Samantha S Iddamaldeniya1, Nalinie Wickramasinghe1, Ira Thabrew2, Neelakanthi Ratnatunge3, Mayuri G Thammitiyagodage4
1Department of Biochemistry, Faculty of Medical Sciences, University of Sri Jayewardenepura, Gangodawila, Nugegoda, Sri Lanka
2Department of Biochemistry and Clinical Chemistry, Faculty of Medicine, University of Kelaniya, Talagolle Roaf, Ragama, Sri Lanka
3Department of Pathology, Faculty of Medicine, University of Peradeniya, Peradeniya, Sri Lanka
4Animal Centre, Medical Research Institute, Colombo 08, Sri Lanka
DOI: 10.1186/1477-3163-2-6


Background: A decoction comprised of Nigella sativa seeds, Hemidesmus indicus root and Smilax glabra rhizome is used to treat cancer patients in Sri Lanka. However, the anti-carcinogenic properties of this decoction have not been experimentally confirmed. The purpose of this study was to determine whether the above decoction could protect against chemically induce hepatocarcinogenesis.
Methods: The effects of this decoction on diethylnitrosamine (DEN) induced hepatocarcinogenesis were examined in male Wistar rats using the medium term bioassay system of Ito, based on a 2-step model of hepatocarcinogenesis. Rats were randomly divided into 6 groups of 10 each. Groups 1 to 4 were injected with DEN (200 mg/kg) to initiate carcinogenesis. Twenty-four hours later groups 1 and 2 were administered the decoction at 4 g/kg body weight/day (dose 1) and 6 g/kg body weight/day (dose 2), respectively. Group 3 and group 4 were given distilled water instead of the decoction and a suspension of garlic powder (20 g/kg body weight/day) in distilled water (positive control), respectively. Group 5 and 6 were injected with normal saline and twenty-four hours later group 5 was given distilled water (normal control) while group 6 was given decoction dose 2 (decoction control). Oral feeding continued for two weeks after which all rats were subjected to 2/3 partial hepatectomy to promote carcinogenesis. Oral feeding continued for eight more weeks. At the end of the 10th week, rats were sacrificed and samples of livers taken for immunohistochemical studies.
Carcinogenic potential was scored by comparing the number, area and staining intensity of glutathione S-transferase placental form (GST-P) positive foci and the number of cells/cm 2 of the positive foci in the livers of the six groups of rats.
Results: The number and area of DEN-mediated GST-P positive foci, number of cells/cm 2 of foci and staining intensity of the foci were significantly (P > 0.001) reduced by the decoction and garlic in the order dose 2 = garlic >dose 1.
Conclusion: Overall results indicate that the decoction comprised of N. sativa , S. glabra and H. indicus has the potential to protect rat liver against DEN induced hepatocarcinogenesis.