Nf-kappa B Trapping Odn Inhibition Of Ectopic And Eutopic Endometrial Secretion Of Rantes And Chemokine Mononuclear Cells

PartⅠComparison of RANTES Expression in Ectopic,Eutopic and Normal EndometriumObjective:To explore the major factor of enhancing monocyte recruitment into the peritoneal cavity.Methods:The stromal cell of ectopic implants was exposed to various concentrations(5,10,20 ng/ml)of IL-1βand the 10 ng/ml of IL-1βwas cocultured with ectopic implants,eutopic and normal endometrium.Concentration of RANTES in the cell culture supernatant was measured with a sandwish ELISA. Results:The 10 ng/ml of IL-1βBelicited the highest secretion of RANTES in ectopic implants.The RANTES secreted by the stromal cell of ectopic implants was increased significantly than eutopic and normal endometrium(P＜0.01).After 60h,the stromal cell of eutopic endometrium secreted much more RANTES than normal endometrium (P＜0.05).Conclusion:These findings are consistent with a feed-forward inflammatory loop whereby IL-1βfrom activate macrophages can lead to RANTES production by ectopic implants and further monocyte chemotaxis.We postulate that this pathway is activated in endometriosis and the bioactivity of eutopic endometrium was different from normal endometrium. PartⅡThe Chemotactic Contribution of RANTES to Monocyte in EndometriosisObjective:To estimate the contribution of RANTES to the total monocyte chemotactic activity in the endometriosis.Methods: Conditioned media from IL-1βtreated ectopic,autologous eutopic and normal endometrial stromal cells were used for the chemotactic assay in a Boyden chamber.Results:The monocyte chemotactic activity in conditioned media of ectopic stromal cells significantly increased compared to autologous eutopic and normal endometrial stromal cells. The 4μg/ml of anti-RANTES antibody almost completely abolished the chemotactic activity of RANTES in the medium and inhibited 62%of the chemotactic activity in conditioned medium from ectopic endometrial stromal cells and55%from autologous eutopic endometrial stromal cells. Conclusion:We established that RANTES accounts for the majority (62%)of the monocyte chemotactic activity in ectopic endometrial stromal cells conditioned media and 55%of that activity in autologous eutopic endometrium.Other monocyte chemokines are likely to contribute the remaining activity of macrophage recruitment. PartⅢThe Secretion of RANTES Depressed by NF-κB decoy ODNs and Changes of its Chemotactic Activity to Monocyte in the EndometriosisObjective:To estimate the effect of NF-κB decoy ODNs inhibiting the secretion of RANTES in the endometriosis and changes of its chemotactic activity to monocyte.Methods:NF-κB decoy ODNs infect the ectopic and eutopic endometrial stromal cells with liposome2000 and then coculture with IL-1βfor 4,8,12,24,36h.Concentration of RANTES in the cell culture supernatant was measured with a sandwish ELISA.Check the chemotactic activity of RANTES in a Boyden chamber.Results:After infection with NF-κB decoy ODNs,the RANTES secreted by eutopic endometrial stromal cells signifinately decreases compared with non-infections(p＜0.05).RANTES to the total monocyte chemotactic activity in infection-teams is signifinately decreases compared with non-infections(p＜0.05).Conclusion:The NF-κB decoy ODNs can decrease the secretion of RANTES in endometriosis and could destroy the inflammatory feedbacks in the development of EMS.