ABSTRACT

‘Triphala' is an age old commonly used Ayurvedic powdered preparation in Indian systems of medicine. In this work, we examined the ability of Triphala to protect against the genotoxicity induced by antineoplastic drug Cyclophosphamide. Micronucleus tests with mouse bone marrow cells by Chromosomal aberration and micronucleus assay were used to examine the effects of Cyclophosphamide. When the comparison between each group was performed for the incidence of percentage chromosomal aberration, it was found that Triphala was non-genotoxic and protective against Cyclophosphamide induced chromosomal aberration. Cyclophosphamide significantly increased the incidence of total percentage aberration (P<0.05). Pre-treatment with Triphala significantly protected against Cyclophosphamide induced chromosomal aberration (P<0.05). The effect of Triphala on Cyclophosphamide induced micronucleus induction was assessed using effect on PCE and NCE. Cyclophosphamide treatment increased total micronucleated PCE significantly (P<0.05). Pre-treatment with Triphala significantly protected against cyclophosphamide induced micronucleus (P<0.05). These results suggest that, Triphala rasayana could be a useful dietary supplement of antineoplastic drug with natural antioxidant activity for cancer patients as it produce a protective effect against genotoxicity.

INTRODUCTION

MATERIALS AND METHODS

-Animals

-Chemicals

-Experimental Protocol

Chromosomal Aberration Assay

Injected Colchicines I.P after 24 hr of chemotherapy.Rats were sacrificed by cervical dislocation after 2 hr of colchicines injection. Animal Dissected and femur bone was excised.Bone marrow was aspirated by flushing with normal saline in the centrifuge tube.Flushed the suspension in the tube properly to get good cell suspension.Centrifuged the suspension for 10 min at 220 rpm and Supernatant discarded.Pellet was treated with pre-warmed (37ºC) 57 % KCl on Vortex shaker.Left above suspension in a water bath (37ºC) fro 20 min.Centrifuged and supernatant discarded.Pellet was treated with freshly prepared cornoy's fixative on Vortex shaker.Centrifuged and supernatant discarded.Above step of treatment with Cornoy's fixative was repeated 3 times to get debris free white pellet.To pellet added Cornoy's fixative (quantity sufficient) to get a good cell suspension.Made the slide with Air Drop Method.Stained (Giemsa's for 3 min. and DDW for 1 Dip) and observed under microscope in 40X and than in 100X magnifications.Number of cells having aberration and the particular aberrations were scored (Total 100 metaphase were counted).

Micronucleas Assay

Bone marrows from both femurs were suspended in Bovine Serum Albumin and then

Smears on clean glass slides were prepared. Slides were air-dried and fixed in methanol. Slides were stained in 10% Giemsa's and coded for a blind analysis. The incidence of micronuclei (MN) was observed in 1000 PCE for each animal.

RESULTS

CONCLUSION

ACKNOWLEDGEMENTS

REFERENCES