Protective effect of DCTN (trans-dehydrocrotonin) against induction of micronuclei and apoptosis by different mutagenic agents in vitro

The use of medicinal plants to combat diseases has increased in the last years despite the little information available with regard to the possible health risks they represent. The aim of the present study was to determine in vitro the possible clastogenic, apoptotic and cytotoxic effects of the active principle of Croton cajucara, trans-dehydrocrotonin (DCTN), and determine its protective effect against three mutagenic agents using the micronucleus test (MN) and apoptosis index in CHO-K1 cells. Three DNA damage inducing agents were utilized in the clastogenicity and anticlastogenicity tests (methylmethane sulfonate (MMS), mitomycin C (MMC) and doxorubicin (DXR); a negative control (PBS) and solvent control were also included. DCTN at concentrations of 400, 320, 240, 160 and 80 mu M did not show clastogenic activity in cultured CHO-K1 cells in the micronucleus test, did not induce apoptosis and showed negligible cytotoxicity in all cases. DCTN at concentrations of 240 and 400 mu M was tested for protective activity using three treatment protocols in relation to positive controls: pre-treatment, simultaneous treatment and post-treatment. The micronucleus test showed a protective effect for DCTN which varied among the different treatment protocols and with regard to the different DNA damage inducing agents. In the apoptosis test, DCTN was seen to have a protective effect under the following conditions: (I) at both concentrations in relation to MMS, in all three treatment protocols; (II) at both concentrations against damage caused by MMC with pre-treatment and at the higher concentration with simultaneous treatment; (III) at both concentrations against DXR with simultaneous treatment. Therefore, DCTN itself is not a clastogenic or cytotoxic substance, and does not induce apoptosis the in vitro system used. These results together with findings reported for DCTN in vivo, support the indication of this active principle at these concentrations for therapeutic use. (c) 2007 Elsevier B.V. All rights reserved.