Investigation To The Expression Of Nrf2 In Glandular Epithelium Of Adenomyosis

Background:Adenomyosis(AM)is a common disease that seriously affects the health of women.Althrough the specific pathogenesis is unclearly,estrogen stimulation and local inflammatory injury are risk factors of this disease.Recently,the incidence of women in childbearing period has been increased year by year.Meanwhile,the age of onset has getting younger and younger.Therefore,the conservative treatment of adenomyosis patients with fertility demands has become a difficult clinical situation.Currently,the accepted pathogenesis of adenomyosis involves the growth and invasion of glandular cells,dysfunction of uterine smooth muscle,stromal cells invagination and so on,among them the invasion and migration of endometrial cells to muscular layer is the key step for occurrence and development of adenomyosis.Matrix metalloproteinases(MMPs)play an important role in the invasion and migration mechanism of adenomyosis.Literature reported that MMPs family can promote invasion and migration of glandular cells by degrading extracellular matrix components.Many studies confirmed that MMP9 expression was abnormal in adenomyosis.Up-regulation of MMP9 expression can promote the invasion of endometrial cells.However,the regulation mechanism and cellular signaling pathway of MMP9 are not clear.Nuclear Factor-E2-related factor 2(Nrf2)is the core protein in the oxidation-reduction regulation mechanism,and participates in various kinds of inflammation stress and degeneration related diseases,as well as the pathogenesis and progression of malignancies.Recent research findings showed Nrf2 was closely related to the invasion and migration of endometrial cancer cells.However,as a chronic inflammation and oxidative stress related disease,it has not been reported that if there is Nrf2 paticipation in the pathogenesis of adenomyosis.It is not clear whether there is a correlation between Nrf2 and MMP9 expression in local tissues of adenomyosis.Our study intended to collect clinical specimens,then use various methods to exam Nrf2 expression levels in the endometria of adenomyosis,clarify the role of abnormal Nrf2 expression in the pathogenesis and progression of adenomyosis,and activate or inhibit Nrf2 expression in vitro to observe the changes of MMP9 and cell migration ability,further illuminate the molecular mechanism of Nrf2 involved in the pathogenesis of adenomyosis.Objective:The purpose of this study was to investigate the expression of Nrf2 in endometrial tissue and its correlation with adenomyosis,and to explore the molecular mechanism of Nrf2 involvement in the pathogenesis of adenomyosis in vitro.Materials and Methods:Collected 20 cases of adenomyosis as well as 20 stage Ia cervical cancer patients during June 2015 to February 2016 in our hospital.The ages between groups were matched and each had 10 proliferatative and 10 secretory endometria.2*2 cm2 endometrial tissues on the posterior uterous wall were removed during the operations.The localization expression of Nrf2 in endometrium was detected by IHC;Differences of Nrf2 mRNA and protein expression levels in adenomyosis and normal endometrial tissues were measured by real-time PCR and Western Blot.The next step was to culture primary endometrial glandular epithelial cells,build cell models with Nrf2 protein over-and poor-expression,test MMP9 protein expression level and the change of cell migration ability using Real-time PCR,Western Blot methods and cell scratch test.Results:1.Endometrial tissue had Nrf2 protein expression.The expression levels of Nrf2 protein in the eutopic and ectopic adenomyosis endometria were significantly higher than that of normal endometrium(P<0.05);Nrf2 expression is present in endometrium epithelial cells.2.There was no significant difference in the expression level of Nrf2 mRNA in endometrial tissues compared with normal endometrial tissues(P>0.05);The expression level of Nrf2 protein in adenomyosis endometrium was significantly higher than that of normal endometrium(P<0.05).3.After the expression of Nrf2 increased in endometrial gland epithelial cells,MMP9 mRNA and protein expression levels raised up significantly,and cell migration enhanced statistically(P<0.05);whereas after the expression of Nrf2 decreased in endometrial glandular epithelial cells,MMP9 mRNA and protein expression levels fell down significantly,and the cell migration rate declined statistically(P<0.05).Conclusions:1.Endometrial tissue had Nrf2 protein expression.The elevated expression level was closely related to the pathogenesis of adenomyosis.2.Nrf2 protein can promote cell migration by up-regulating the expression of MMP9 in endometrial epithelial cells,thereby participating in the pathogenesis and progression of adenomyosis.