Figure 4.

(A) Bioassay-guided fractionation of G. inflata extract. The crude extract of G. inflata was fractionated by countercurrent chromatography. The different G. inflata fractions were tested for their estrogenic properties on both ERα and ERβ models. Fractions 8 and 10 displayed significant activity on both the ERα and ERβ models. Fraction 8, with the highest mass yield at 2.2% w/w crude extract (see Supporting Information), was further processed by semipreparative HPLC to isolate and identify four major compounds, namely, 8-prenylapigenin (3), licochalcone C (5), licochalcone A (6), and abyssinone II (4). (B) Induction of differential estrogenic activity with the isolated compounds from the active bioassay-guided fractions (A) in an alkaline phosphatase activity induction assay in Ishikawa cells (ERα) and in an ERβ-ERE-luciferase assay in MDA-MB-231/β41 cells. The methods for the Ishikawa and ERE-luciferase assays are described in the Experimental Section. The data represent the means ± SEM of three independent determinations.