Huperzia saururus effects over eNOS expression and cell toxicity

BIRRI M; PELLIZAS C; DONADIO AC; AGNESE AM

Simposio; XXII Simposio de Plantas Medicinais do Brasil; 2012

Introduction: As WHO promoted the study of plants used in enthomedicine avoiding dangerous practices and promoting those that are safe (Bermúdez. Interciencia. 30(8), 453-59, 2005) we investigated Huperzia saururus (Lam.) Trevis. (Lycopodiaceae) (Hs), used in Argentina as afrodisiac (Amorín. Farmacobotánica. 16, 3-6, 1977) with the objective of confirm or reject scientifically this use. The aim of this work was to assay Hs citotoxicity and eNOS protein expression in view of its potential therapeutic use. Experimental: 5g of in shade air-dried and crushed plant material (PM) were boiled in 100mL of H2O (FA VII Ed, 2004) during 20 min. Obtained decoction (D) was filtered and concentrated to get the yield. U937 or mononuclear cells from peripheral blood of a healthy donor (MPBC) were incubated in RPMI-SBF 10%, at 37°C during 24h. Then, the cells were incubated with D at 50, 100, and 500µg/mL in RPMI-Calf serum 5% during 6, 24, and 48h. Cell citotoxicity was determined using a tetrazolium based colorimetric assay (MTT), (Garay. Nanomedicine. 5(4), 443-51, 2009). Absorbaces (Abs) were read at 595 and 650nm. The same assay was done using an ethanolic extract (E) prepared by maceration of 5 g of MP in 20mL of EtOH, at 10, 50, and 100µg/mL. As control (C), U937 and MPBC cells were cultured without D or E. Abs statistic analysis was performed applying Chauvenet criteria. eNOS expression was determined by Western blot, using cell extracts of MPBC incubated with D during 6, 24 and 48h. Results and Discussions:  Hs citotoxicity Taking C absorbance as 100% of viability, Hs extracts (D and E) did not show a citotoxicity effect in U937 and MPBC cultures.  Hs eNOS expression An increase in eNOS expression was observed in MPBC cultures at 24h, with a strong signal using D 50µg/mL. Conclusions: In view that toxicity is considered when problem values are lesser than 95% of C, the obtained results show that both extracts are not toxic in the assayed conditions. The increases of eNOS expression taken as an indirect measure of nitric oxide production, would endorse the use of Hs as an aphrodisiac.