Gui Shao Di Huang Wan Promotes Angiogenesis and Regulates Oestrogen Receptor α

The formula Gui Shao Di Huang Wan (GSDW) is used frequently in Chinese Medicine to treat female infertility. This study aims to investigate some of the possible mechanisms of action of GSDW using in vitro bioassays of angiogenesis in Human Uterine Microvascular Endothelial Cells (HUVEC) and Human Uterine Microvascular Endothelial Cells (HUtMEC) and ovarian steroid receptor expression in a human endometrial cell line (Ishikawa). Aqueous extracts of GSDW and its component herbs were tested for pro-angiogenic activity using both HUVEC and HUtMEC 2D differentiation assays performed on Matrigel and effects on HUVEC proliferation using the MTT assay. Effects on the expression of Oestrogen Receptor α (ERα) and Progesterone Receptor β (PRβ) in Ishikawa cells were determined using immunoblotting. All analysed parameters of differentiation were increased by GSDW in both the HUVEC and HUtMEC mesh. Furthermore, measures of total length, segment number, junction number were affected by some but not all component herbs. The MTT assay showed an increase in proliferation of HUVECs at concentrations of GSDW between 0.68 and 5.47 µg/mL at 48 and 72 hours. In Ishikawa cells, down-regulation of ERα and up-regulation of PRβ was seen after 48 hours incubation with 4 of the 8 herbs in the formula. The findings in this study demonstrate that GSDW has the potential to affect key parameters (vascular, sex hormone receptor expression) in vitro. This offers a potential mechanism by which these herbs may enhance fertility through improved endometrial receptivity and pregnancy rates.