Metformin reverses infertility in a mouse model of endometriosis: unveiling disease pathways and implications for future clinical approaches

Research question: Does metformin reverse endometriosis-associated infertility? Design: Endometriosis was induced by transplanting uterus fragments from B6CBAF1 mice into recipients of the same strain. The mice were divided into groups: endometriosis (End, n = 24), sham-operated (Sham, n = 12), endometriosis with metformin (0.5mg/ml) orally administered for 3 months (EndMet, n = 21) and sham-operated metformin-treated (ShamMet, n = 16). Implant growth was monitored using ultrasonography. Fibrosis was computer-assisted quantified in Masson's trichrome-stained sections of eutopic (EuEnd) and ectopic (EcEnd) endometrium. PCNA, CYP17a1, F4/80 and galectin-3 were analysed by immunofluorescence and western blotting, and NFkB, GPX-1 and HO-1 only by western blotting. Statistical significance was set at P <0.05. Results: The endometriosis model was successfully established. The End groups showed lower fertility rates than sham-operated mice (P = 0.0034), whereas metformin treatment increased the number of fetuses per pregnant mouse (P = 0.0295), restoring fertility to control levels; it also slowed implant growth and vascularization. Metformin also restored PCNA expression and fibrosis levels to those of non-treated EuSham mice. PCNA expression decreased in pregnant mice (P <0.0178). Metformin diminished CYP17a1 expression in EcEnd versus EuEnd non-treated tissues and conversely up-regulated F4/80 in EuEnd tissue (P <0.0170), and galectin-3, NFkB and the antioxidant enzymes HO-1 and GPX-1 in EcEnd tissue (P <0.0293), in non-mated mice. Conclusions: These results indicate that application of metformin can alleviate oxidative stress and mitigate fibrosis in endometriosis lesions in a murine model of endometriosis, which highlights metformin's potential as a pharmacological intervention for improving infertility in endometriosis.