The aqueos extract of Smilax campestris Griseb. diminished resistant acid phosphatase positive cell number induced by RANKL, and increase apoptosis on RL-induced osteoclast-like cells, acting through ER alpha.

MANGONE F; SALAVERRY L; SOTELO A; PARRADO CECILIA; REY-ROLDÁN ESTELA; CASTRO MS; CANELLADA AM

Mar del Plata

Congreso; Reunión Conjunta de Sociedades Biomédicas; 2017

Sociedad Argentina de Inmunología

Smilax campestris Griseb. (SM), is the most widely distributed Smilax specie in the north of Argentina and it is used as antirheumatic folk 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 that the treatment of RAW cells with aqueous extract of SM diminished the in vitro osteoclastogenesis induced by RANKL. Also, NFATc1 nuclear translocation was inhibted by SM in a dose-dependent manner, acting at level of oestrogen receptors alpha (ER alpha). This work aimed to analyze the mechanisms involved in the inhibitory effect of SM on the in vitro osteoclastogenesis induced by RANKL. Specific objectives developed were: 1) to compare the effect of SM and oestrogen on the differentiation to tartrate resistant acid phosphatase (TRAP) positive cells, 2) to determine whether SM induce apoptosis 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 or absence 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 days for TUNEL assay. To analyze the role of ER, RAW cells were incubated for 1 h with ER alpha or ER beta antagonist previous to SM treatment. SM 1000 ng/ml inhibited the RANKL effect on TRAP expression and MMP activity (p