Objectives Estrogen deficiency causes various health problems in postmenopausal women, including osteoporosis. Phytoestrogen emerged as a potential alternative of estrogen with minimum side effects. The aims of this study were to analyze the metabolite profiling results of various extract of Chyrsophyllum cainito L. leaves, which contain phytoestrogen, through in silico study against 3OLS protein, an X-ray protein of ERβ, so it can predict the types of the phytoestrogen contents which have antiosteoporosis property. Methods In silico analysis was carried out for the compounds from the metabolite profiling data of C. cainito leaves from our previous study. The structure compounds from metabolite profiling results of various extract of C. cainito leaves were prepared with Avogadro 1.0.1 software, molecular docking was done using PyRx 0.8 software, and Biovia Discovery Studio Visualizer 2016 software was used to visualize the structure of compounds against 3OLS protein. The physicochemical characteristics of the compounds were analyzed using the SwissADME web tool. Results From in silico studies, it was known that there were total 11 compounds in C. cainito leaves that predicted as phytoestrogens which have ERβ agonist properties against 3OLS protein. The ERβ agonist was a compound that has parameters similar to 17β-estradiol in its interaction with 3OLS protein, which has a pharmacophore distance of 10.862 Å, and binding to amino acids His 475 and Glu 305 or Arg 346 at receptor-ligand docking simulation. Conclusions C. cainito leaves contain 11 compounds that are predicted to be phytoestrogens with ERβ agonist properties, which is responsible for antiosteoporosis activity.

Neuroinflammatory is a pathological condition in which neuronal cells experience progressive loss of function, one of which is due to estrogen deficiency. Phytoestrogens are compound that has structure and function similar to 17β-estradiol, the most potent natural form of estrogen. Semanggi (Marsilea crenata Presl.) is known to contain phytoestrogens which can replace the function of 17β-estradiol in maintaining organ homeostasis, so that is potential to be developed as an anti-neuroinflammatory drug. This study was aimed to prove the anti-neuroinflammatory effect of the n-hexane and ethyl acetate fraction of Semanggi leaves on HMC3 microglia cells, by measuring the expression of major histocompatibility complex II (MHC II). Both fractions with a concentration of 62.5; 125; and 250 μg/ml were administered to HMC3 microglia cells which had been induced by interferon gamma (IFN-γ) for 24 hours. Measurement of MHC II expression were using immunocytochemistry (ICC) methods and confocal laser scanning microscopy (CLSM) instruments. The results showed that n-hexane fraction could reduce MHC II expression at concentrations of 250 μg/ml with p<0.05, and showed a non-monotonic dose response (NMDR) pattern, while ethyl acetate fraction could reduce MHC II expression at all concentrations with p<0.05. Based on the results of the correlation test, the best anti-neuroinflammatory activity on HMC3 microglia cells is showed by the ethyl acetate fraction of Semanggi leaves.