THE AQUEOUS EXTRACT OF SMILAX CAMPESTRIS DIMINISHED RESISTANT ACID PHOSPHATASE POSITIVE CELL NUMBER INDUCED BY RANKL, AND INCREASE APOPTOSIS ON RL-INDUCED OSTEOCLAST- LIKE CELLS, ACTING THROUGH ER ALPHA

SOTELO AGUSTINA; DIAZ AILEN; CANELLADA ANDREA; SALAVERRY LUCIANA; ALMOZNI BRENDA; CASTRO MARISA; MANGONE FRANCO; PARRADO CECILIA; REY-ROLDAN ESTELA

Buenos Aires

Congreso; Reunion conjunta de Sociedades de Biociencias; 2017

Smilax campestris Griseb. (SM), is the most widely distributedSmilax specie in the north of Argentina and it is used as antirheumaticfolk medicine due to its anti-inflammatory and anti-oxidant properties.Our lab is studying the possible therapeutic effect of SM on oestrogen-induced bone resorption. Previously, we demonstrated thatthe treatment of RAW cells with aqueous extract of SM diminishedthe in vitro osteoclastogenesis induced by RANKL. Also, NFATc1nuclear translocation was inhibted by SM in a dose-dependent manner,acting at level of oestrogen receptors alpha (ER alpha). Thiswork aimed to analyze the mechanisms involved in the inhibitoryeffect of SM on the in vitro osteoclastogenesis induced by RANKL.Specific objectives developed were: 1) to compare the effect of SMand oestrogen on the differentiation to tartrate resistant acid phosphatase(TRAP) positive cells, 2) to determine whether SM induceapoptosis on RANKL-induced osteoclast-like cells by TUNEL assay,and 3) to analyze the effect of SM and oestrogen on the metalloproteinase-9 (MMPs) activity by Zymography.RAW cells were cultured with RL (100 ng/mL) in the presence orabsence of SM (10-100-1000 ng/mL) and oestrogen (10-8, 10-7,10-6 M) during 5 days for TRAP and Zymography, or during 2 daysfor TUNEL assay. To analyze the role of ER, RAW cells were incubatedfor 1 h with ER alpha or ER beta antagonist previous to SMtreatment. SM 1000 ng/ml inhibited the RANKL effect on TRAP expressionand MMP activity (p<0.0001) similarly to inhibition inducedby oestrogen. Inhibitory activities for TRAP of SM were reversedby ER alpha antagonist (p<0.01). In the apoptosis studies, we observedthat SM 10 ng/mL induced the cell apoptosis (p<0.0001). ERalpha antagonist revert the apoptotic effect of SM (p<0.001).We conclude that aqueous extracts of SM are capable to significantlydiminish TRAP expression and induce apoptosis on RL-inducedosteoclast-like cells, acting through ER alpha.