Cytogenotoxic effects of a characterized aqueous extract of baccharis articulata (lam.) pers. (Asteraceae) on normal cells.

ESCOBAR F; SABINI M; TORRES C; RESER AL; COMINI LR .; SABINI L

Bento Goncalves

Simposio; XXII SIMPOSIO DE PLANTAS MEDICINAIS DO BRASIL; 2012

Comite organizador del evento

Introduction: Cold aqueous extract of Baccharisarticulata (Ba-CAE) induces the death of human peripheral blood mononuclear cells (PBMCs) and exerts mutagenic effects on mice at 6h post-injection (p.i). The aimwas to investigate whether the PBMCs death is due to apoptosis, and whether this extract exerts mutagenic effects on mice at 24 and 48 hp.i. In addition, Ba-CAE was chemically characterized. Materials and methods: PBMCs were exposed to extract (10, 20, 40, 80, 160, 320, 640 and 1280 microgramos/mL) for 18-24 h. Cell viability was determined by trypan blue dye exclusion method. Apoptosis was determined by Hoechst 33258 staining, TUNEL, and DNA fragmentation analysis by agarose gel electrophoresis. BALB/c mice were injected with extract (1800, 900 and 450 mg/kg) and sacrificed at 24 and 48 h p.i. Bone marrow samples were used to assess chromosome mutations by the micronucleus test. The main metabolites in the Ba-CAE were identified and quantified by HPLC. Results: A dose-dependent toxicity of Ba-CAE was demonstrated on PBMCs (CC50=165mg/mL). Hoechst staining showed several apoptotic figures. The percentage of TUNEL-positive per 400 cells was: control: 7+1%, cells treated: 10microgramos/mL(8+1%), 20 microgramos/mL(9+3%), 40  microgramos/mL(43+11%), 80  microgramos/mL(58+18%), 160  microgramos/mL(62+13%), 320  microgramos/mL(68+20%), 640  microgramos/mL(73+10%), 1280  microgramos/mL(88+13%); p<0.01. The agarose gel electrophoresis showed the typical DNA laddering in cells treated with Ba-CAE. Both at 24 and 48h, the extract increased the frequency of micronuclei in polychromatic erythrocytes (Tukey test, p<0.05). HPLC analysisdemonstrated the presence of luteolin(1.96 ± 0.27%), acacetin (1.12 ± 0.14%) and chlorogenic acid (0.29 ± 0.05%). Conclusion: Ba-CAE induced apoptosis and mutagenic effects on normal cells.The phytotherapy use of this medicinal herb should be limited as it may cause serious damage to cells when used improperly. Funding:CONICET,PICTOR 8/20325 BID 1728/OC-AR and SeCyT, UNRC.