Reduced endometrial glycolysis concomitant with increased lesional fibrosis in patients with adenomyosis who complained of heavy menstrual bleeding

Research question: What role, if any, does the extent of lesional fibrosis play in impaired glycolysis leading to adenomyosisassociated heavy menstrual bleeding (ADM-HMB)? Design: Forty-eight patients with ADM-HMB were recruited, among them 25 reported moderate to heavy bleeding (MHB), and the remaining 23, excessive bleeding (EXB). The full-thickness uterine tissue columns were processed for Masson trichrome staining and immunohistochemistry analyses. The expression levels of HIF-1a, GLUT1, HK2, PFKFB3 and PKM2 proteins that are critically involved in glycolysis in endometrial epithelial cells cultured on substrates of different stiffness, and the levels of glycolysis were quantitated. A mouse experiment with induced adenomyosis and simulated menstrual bleeding was conducted to assess the effect of adenomyosis on immunoexpression of proteins involved in glycolysis and inflammation as well as on endometrial repair and bleeding. Results: The endometrial staining of HIF-1a, GLUT1, HK2, PFKFB3 and PKM2 was significantly lower in the EXB group as compared with MHB patients, concomitant with higher extent of fibrosis. The expression of HIF-1a, GLUT1, HK2, PFKFB3 and PKM2 was significantly reduced when endometrial epithelial cells were cultured in stiff substrate, concomitant with reduced glycolysis. Mice with induced adenomyosis had reduced immunoexpression of Hif-1a, as well as those proteins each of which plays a vital, rate-limiting role in different steps of the glycolysis pathway, such as Glut1, Hk2, Pfkfb3 and Pkm2, and elevated fibrosis in endometrium, concomitant with disrupted endometrial repair and more bleeding. Conclusions: Lesional fibrosis results in reduced endometrial glycolysis in eutopic endometrium and subsequent imbalance in pro-inflammatory and anti-inflammatory response, leading to ADM-HMB.