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), 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 citotoxicity and to probe the activity of one aqueous extract as eNOS producer.
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 cells were incubated in RPMI-SBF 10%, at 37°C during 24h. Then, cultured media was replaced by RPMI-Calf serum 5%, and the cells incubated with increasing D concentrations of 50, 100, and 500µg/mL during 6, 24, and 48h. As control (C), U937 cells were cultured without D. 4h before to the end time of incubation 50µL of MTT (5mg/mL), (Garay. Nanomedicine. 5(4), 443-51, 2009) were added. Next, medium was removed and the cells were lised with DMSO. 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. An equivalent experiment was performed using mononuclear cells obtained from peripheral blood of a healthy donor (MPBC). The
statistic analyisis of the absorbances was evaluated applying Chauvenet criteria.
The same methodology was used to measure the increase in the eNOS production by D using MPBC. The difference was that at the end of the incubation time the cells were lised with sample buffer and then analysed by western blot.
Results and Discusions: C absorbance represents 100% of de viability.
|
Viability (%) | ||||||
|
Time (h) |
D (µg/mL) |
E (µg/mL) | ||||
|
500 |
100 |
50 |
100 |
50 |
10 | |
|
U937 | ||||||
|
6 |
117 |
175 |
169 |
194 |
241 |
164 |
|
24 |
114 |
127 |
188 |
165 |
216 |
224 |
|
48 |
118 |
188 |
181 |
186 |
254 |
226 |
|
CMDS | ||||||
|
6 |
108 |
106 |
124 |
111 |
98 |
82 |
|
24 |
98 |
96 |
86 |
101 |
136 |
113 |
|
48 |
95 |
98 |
102 |
97 |
114 |
126 |
On the other hand, D showed an increase of 130, 40 and 55% in comparison to C at 50, 100 and 500µg/mL respectively 24 h after incubation.
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. In relation with the D activity, the increase in the eNOS production observed would indicate a correlation with the claimed activity in the ethnomedicine.
