Study On Differential Expression Of MANF,iNOS And CD206 In Women With Endometriosis

Objective: To understand the expression levels of astrocyte-derived neurotrophic factor(MANF),inducible nitric oxide synthase(i NOS)and cluster of differentiation206(CD206)in eutopic endometrium and ectopic endometrium of patients with endometriosis(EMT),and to explore whether the expressions of MANF,i NOS and CD206 in different endometrium tissues are different.Methods: Twenty patients with ovarian endometriosis who underwent ovarian endometriosis cystectomy from September 2017 to September 2018 in our hospital were enrolled in the experimental group.Twenty patients with infertility due to simple fallopian tube disease were included in the control group.In the experimental group and the control group,the Eutopic endometrium(EUE)was collected through the disposable uterine tissue suction tube.The ectopic endometrium(ECE)of the experimental group was collected by removing the intraoperative ectopic cyst wall from the intraoperative detachment.Immunohistochemistry,Western blotting,real-time fluorescent quantitative PCR and immunofluorescence were used to detect the expression differences of MANF,i NOS and CD206 in different endometrial tissues.Results: 1.Immunohistochemical results: MANF,i NOS and CD206 were expressed in normal endometrium,endometrium in situ and endometrium in ectopic tissue.The positive rates of MANF in normal endometrium,in situ endometrium and ectopic endometrium were 20.00%,35.00%,and 70.00%,respectively.INOS were 20.00%,25.00% and 60.00%.CD206 was 25.00%,30.00% and 60.00%.The expression of MANF and i NOS in the ectopic endometrium was higher than that in the incumbent endometrium(?2=4.912,P=0.027 and ?2=5.013,P=0.025).The expression of MANF,i NOS and CD206 in the ectopic endometrium was higher than that in the normal endometrium(?2=8.120,P=0.004;?2= 6.667,P = 0.010;?2= 5.013,P = 0.025).2.Western blot results: The relative gray values of normal endometrium,in situ endometrium and ectopic endometrium were(1.03 0.17),(1.16 0.16)and(1.44 0.09).INOS were(0.54 0.12),(0.71 0.05)and(1.24 0.10).CD206 was(0.20 0.04),(0.29 0.05)and(1.08 0.11).The expression of i NOS and CD206 in the ectopic endometrium was higher than that in the in situ endometrium(P<0.05).The expression of endometrium MANF,i NOS and CD206 was higher than that of normal endometrium in the control group(P<0.05).3.q PCR results: The normal endometrial CT value was(37.44±1.37)and the ?-actin CT value was(18.65±1.11).The eutopic endometrial MANF CT value was(36.82±1.38),the ?-actin CT value was(18.44±1.04);the ectopic endometrial CT value was(40.00±0.00),and the ?-actin CT value was(24.52±4.89).The number of cycles of MANF m RNA in the three groups of data was greater than 35,and the results were meaningless.4.The results of immunofluorescence experiments: co-localization of i NOS and MANF in the eutopic endometrium and ectopic endometrium.In the experimental group,the number of i NOS and MANF double positive cells in the ectopic endometrium was higher than that in the eutopic endometrium(P <0.05);compared with the control group,the number of double positive cells in the ectopic endometrium and the eutopic endometrium was higher than that in the normal endometrium(P<0.05).Co-localization of CD206 and MANF was observed in the eutopic endometrium and ectopic endometrium.The number of ectopic endometrial CD206 and MANF double positive cells in the experimental group was higher than that in the eutopic endometrium(P<0.05);In comparison,the number of double positive cells in the ectopic endometrium and the eutopic endometrium increased(P<0.05).Conclusion: EMT patients have high expression of MANF,i NOS and CD206 in ectopic endometrial tissues.i NOS and CD206 co-localize with MANF in eutopic endometrium and ectopic endometrial tissue of EMT patients,suggesting that ectopic endometrial tissue promotes the recruitment of local macrophages through high expression of MANF,which changes the immune microenvironment and is beneficial to the development of endometriosis.