Steroidogenic activity of methylated resveratrol analogue 3,4,5,4′-tetramethoxystilbene (DMU-212) in human luteinized granulosa cells in a primary three-dimensional in vitro model

Abstract

Abstract Purpose One of the main functions of granulosa cells (GCs) is the secretion of steroid hormones. Resveratrol is a natural polyphenol, known for its beneficial health effects, such as improving reproductive health. However, its application is limited due to poor bioavailability. Methoxy derivative of resveratrol (DMU-212) was shown to be more lipophilic, and consequently more bioavailable. However, since the addition of methoxy groups to the stilbene scaffold was found to make the molecule insoluble in water, DMU-212 was loaded into liposomes. This study aimed to evaluate how the liposomal formulation of DMU-212 (lipDMU-212) alters estradiol and progesterone secretion of human ovarian GCs in a primary three-dimensional cell culture model. Methods DMU-212-loaded liposomes were prepared by thin film hydration followed by extrusion. Cell viability was measured after exposure of GCs spheroids to liposomal formulation of DMU-212 using CellTiter-Glo® 3D Cell Viability Assay. The secretion of estradiol and progesterone was determined using commercial ELISA kits. RT-qPCR was conducted to analyze expression of steroidogenesis-related genes. Results lipDMU-212 was found to significantly increase estradiol and progesterone secretion in a dose-dependent manner by enhancing expression of CYP11A1, HSD3B1, CYP17A1, CYP19A1, and HSD17B1 genes. Furthermore, our study suggests that lipDMU-212 increases the FSH activity. Conclusions This is the first study to describe the steroidogenic activity of liposomal formulation of DMU-212, possibly through increasing the StAR and CYP19A1 expression. These findings suggest that lipDMU-212 might have a beneficial effect in the treatment of disorders related to estrogen deficiency and hyperandrogenism, such as PCOS.