Researches About The Role And Mechanism Of LipoxinA₄ Activating Autophagy In Ectopic Stromal Cells Of Endometriosis By Inhibiting Inflammation

Background:Endometriosis(EMs),it is a gynecological disease that seriously harms women’s physical and mental health and quality of life.The cause of its onset is unclear.More and more studies show that inflammation act a pivotal part in the development of endometriosis.Autophagy is a constitutive catabolic pathway that participates in various pathophysiological processes.Recent studies have shown that some autophagy-related proteins and genes are abnormally expressed in patients with endometriosis,suggesting that autophagy is closely related to the pathogenesis of endometriosis.Lipoxin A4(LXA4)is an arachidonic acid metabolite that act a pivotal part in the inflammatory response.Our previous studies confirmed that LXA4 can inhibit the growth of endometriotic lesions.AKT-mTOR is an important signal transduction pathway,plays a key role in the regulation of inflammatory response and autophagy.Studies showed the AKT-mTOR pathway is abnormally expressed in endometriosis.Objective:The purpose of this paper is to explore whether inflammation and autophagy are linked in endometriosis and what pathways LXA4 uses to regulate inflammation and autophagy to inhibit the occurrence of endometriosis.With a view to providing new ideas for the causes and treatment of endometriosis.Methods:In this thesis,the expressions of autophagy-related proteins LC3B,p62 and autophagosomes in normal endometrial stromal cells and endometriosis ectopic interstitial cells(ESCs)were detected by Western blot and transmission electron microscopy;The effects of inflammation on ESCs proliferation and migration ability were analyzed by CCK-8 and Transwell;The relationship between inflammation and autophagy levels in ESCs was analyzed by Western blot and qRT-PCR,and the effect of inflammation on AKT-mTOR signal transduction.The effect of LXA4 on ESCs autophagy level and the effect of LXA4 on the reduction of autophagy level of ESCs caused by inflammation and the activation of AKT-mTOR signaling pathway were analyzed by Western blot.The expression of IL-1β and autophagy-related proteins LC3B,p62 and key proteins of AKT-mTOR pathway,p-AKT(T308)and p-mTORS2448 in ESCs were analyzed by inhibiting LXA4 receptor AhR.Finally,a BALB/c mouse model of endometriosis was constructed,and the effects of LXA4 on the inflammation and autophagy levels in endometriosis mouse were analyzed by immunohi stochemi stry.Results:1.In ESCs,LC3B is low expressed and p62 is highly expressed;the number of autophagosomes is less than that of normal intimal stromal cells.2.In ESCs,after the induction of inflammation,the cell’s proliferation and migration ability is significantly improved,and the results of autophagy-related protein expressions indicate that the autophagy level is reduced,and the levels of the key proteins of AKT-mTOR pathway,p-AKT(T308)and p-mTOR-S2448 were increased;after induction of autophagy,the inflammation level decreases;After inhibiting autophagy,the level of inflammation increased again.3.In ESCs,LXA4 can reduce inflammation,inhibit AKT and mTOR protein phosphorylation,and increase its autophagy level through its receptor.4.The effect of LXA4 on inflammation and autophagy in endometriotic mouse was explored in vivo.The results showed LXA4 had no significant function on the weight of BALB/c mouse with endometriosis,but it could make ectopic lesions smaller,and LXA4 can reduce the level of inflammation and increase autophagy in mouse with endometriosis.Summary:In ESCs,inflammation is negatively correlated with autophagy,abnormal autophagy is a major incentive for the development of endometriosis.LXA4 can down-regulate inflammation through its receptors AhR,inhibit AKT and mTOR protein phosphorylation,and up-regulate autophagy,thereby inhibiting the occurrence and development of endometriosis.