Evaluation of protective potential of a crude plant extract (Chelidonium majus) against p-dimethylaminoazobenzene induced hepatocarcinogenesis in mice.
Cytogenetics Laboratory, University of Kalyani, West Bengal, India
Aim: To examine if ethanolic whole plant extract of the celandine, Chelidonium majus, used as homeopathic medicine against various liver ailments, has any protective potential against p-dimethylaminoazobenzene (p-DAB) induced hepatocarcinogenesis in mice as revealed from cytogenetical, biochemical, histological and electron microscopical analyses. Methods: Adult, healthy mice, weighing between 18-22 gms and maintained in hygienic conditions (under supervision of the Animal Welfare Committee, Kalyani University), with normal food without protein supplement and water ad libitum, were divided into various sets of 5-10 mice each for each fixation interval: i) Set-I. Mice were chronically fed p-DAB (initiator,@0.06% mixed with diet) and phenobarbital (PB, promoter, @0.05 ml/day) to develop liver tumors in about 60 days; ii) Set-II. Mice fed chronically p-DAB and PB were also fed 0.1 ml of crude plant extract (Chelidonium majus- , collected from HAPCO, Calcutta) daily, iii) Set-III. Mice were fed chronically p-DAB and PB plus dilute ethyl alcohol ("vehicle" of the plant extract) (positive control), and iv) Set-IV. Mice were maintained with normal diet without any kind of treatment (negative control). Different sets of mice were sacrificed at six different intervals, namely, 7, 15, 30, 60, 90 and 120 days after treatment or otherwise. Data of several cytogenetical endpoints like chromosome aberrations, micronuclei and mitotic index of bone marrow cells and sperm head abnormality, biochemical assay of some toxicity marker enzymes like lipid peroxidase (LPO), acid and alkaline phosphatases (ACP, ALP), glutamate oxalo-acetate transaminase (GOT) and glutamate pyruvate transaminase (GPT),at all fixation intervals and histological observations of liver sections through ordinary, scanning and transmission electron microscopies at 60 and 120 days have been critically analyzed in control and treated lots, by following standard current methodologies. Results: In the Set-I and Set-III mice, tumors were found to develop in liver generally from 60 days onward in increasing numbers with the lapse of time. In Set-II mice (receiving the plant extract along with p-DAB+PB), fewer incidences of tumor occurred. Further, all the cytogenetical, biochemical, histological and electron microscopic observations in the different sets of mice positively indicated protective role of the plant extract as compared to controls, leading to some understanding of its possible mechanism of action. Conclusions: This plant extract, even in microdoses, has hepato-protective, anti-tumor, anti-mutagenic, and anti-carcinogenic potentials against p-DAB induced hepatic tumors in mice, which validates its traditional use in many herbal medicines against suspected liver damage.
Paper presented at the International Symposium on Predictive Oncology and Intervention Strategies; Nice, France; February 7 - 10, 2004; in poster session 997 (Chemoprevention).