IL-6 Level And Ultrastructural Change In Endometriotic Tissue

Objectives:1. To explore the alteration of IL-6 level in EOS and eutopic EM at sixth weeks after modeling and the association with pathogenesis of endometriosis.2. To investigate the effects on appearance and IL-6 level of EOS. To explore how Danazol can be used to treat endometriosis.3. To observe the appearance and ultrastructural change of EOS and to provide some cues for pathogenesis of endometriosis.Methods:1. Thirty female rats were divided randomly into three groups: control group (n = 10) , modeling group (n=10) and Danazol-treated group (n=10) .2. Vaginal smears were performed on all rats (modeling group / treated group) before auto-implantation to ensure that endometriosis was induced surgically during estrum.3. At the third week after the initial surgery, each rat was anesthetized and a laparotomy was performed to determine the state of the endometrial explants. The explants was measured during surgery. At the same time treatment was begun, Danazol was administered as a subcutaneous injection at a repeated dose of 12 mg/kg per day for 3 weeks. The modeling groups were given subcutaneous injection of normal saline.4. At the third week after the second surgery, the laparotomy was performed again and the explants were measured, transplant then was excised for detemination of the IL-6 level and ultrastructural examination.5. The excised tissue is weighed accurately and triturated completely together with PBS buffer. Total IL-6 is extracted from the mixture by centrifugation,the concetration of IL-6 is determined by ELISA. Statistical analyses were carriedout by SPSS 10.0 software. P < 0.05 was considered to be statistically significant.Results:1. At the second laparotomy, explants from experimental rats grew well and became larger, EOS produced transparent fluid-filled cysts. Neovascular networks were clearly visible on the surface of transplants. However there were no explants from control rats.2. At the third laparotomy, the volume of endometrial explants from modeling group was not significantly changed when compared with that observed at the second laparotomy (P > 0.05) and transplants appeared similar at last two laparotomies. But significant difference was noted for the size of explants from Danazol-treated group (P < 0.01).3. For modeling group, there is obvious difference between the IL-6 level from eutopic EM and that from EOS at the third laparotomy (P < 0.05) . The IL-6 levels from the ectopic and EOS in Danazol-treated group was significantly higher than those in Danazol-treated group respectively (P < 0.01, P < 0.05).4. Compared with the eutopic EM from control and modeling groups, EOS has discontinuous the basement membrane and vacuolated mitochondrion; after Danazol treatment, eutopic EM have more auto phagosome > bacilliform mitochondrion and more white cells, necrotic celK vacuolated > expanded rough endoplasmic reticulum and auto phagosome in EOS were commonly seen.Conclusions:1. Auto-transplants grow well, transplants from modeling group observed at the third laparotomy wer similar with that observed at the second laparotomy; Three weeks' treatment with Danazol can make EOS atrophy.2. The IL-6 level from EOS in modeling group is lower than that in eutopic and normal EM, three weeks' treatment make the IL-6 level in endometriotic tissue increase significantly, which suggests that the decreased IL-6 plays a vital role in the onset of endometriosis.3. Ultrastucture of EOS from modeling group is different from eutopic EM, Three weeks' treatment with Danazol can make organelle altered and cell necrotized.