In vitro & in vivo evaluation of anti-fibrotic effects of Rg3-enhanced extract and identification of the associated miRNAs in endometriosis

Objective: The potential therapeutic effects of ginsenoside Rg3 on endometriosis and its target miRNAs were identified. Methods: An in vitro study using human endometrial stromal cells (HESCs) obtained from patients with endometriosis and an in vivo study using mouse endometriosis models were designed. HESCs were treated with Rg3-enhanced extract (Rg3E); real-time PCR, microarray profiling, transfection, and western blot were performed. Mouse endometriosis models were developed and supplemented with Rg3E for 8 weeks. Results: RNA levels of Ki-67, Col-1, CTGF, fibronectin, TGF-棺1, MMP2 and 9 were significantly decreased in Rg3E-treated HESCs. Microarray analysis revealed downregulation of miR-27b-3p, which is related to fibrosis modulation. Expression of miR-27b-3p was significantly higher in HESCs from patients with endometriosis than that of controls, and Rg3E treatment significantly decreased its expression. The transfection of miR-27b inhibitor revealed significant decreases in Col-1 and MMP9. The protein expression results from western blot were also comparable. The contraction and migration assay revealed significant reductions in both fibrosis and migration potential in Rg3E-treated HESCs from endometriosis patients. In Rg3E-treated mice, decreases in size and fibrotic character of endometrial lesions were observed. Conclusion: Rg3E effectively alters fibrotic properties of HESCs from patients with endometriosis, which is presumed to be associated with miR-27b-3p modulation.