Changes And Significance Of Th17/Treg Cells Before And After Surgery And GnRH-a Treatment In Patients With Endometriosis

Objective:To detect the proportion of helper 17 cells and regulatory T cells in peripheral blood of patients with endometriosis before and after surgery or treatment with gonadotropin releasing hormone agonist and the changes and significance of the expression levels of cytokines related.Methods:The subjects were selected from patients who had undergone surgery in the department of gynecology,Affiliated Hospital of Qingdao University,from January 2019 to December 2019.The endometriosis group consisted 40 patients with ovarian endometriosis cysts.According to whether treated with GnRH-a one month after laparoscopic surgery,they were divided into two groups,the surgery-only treatment group was 22 cases without medication,and the GnRH-a group was 18 cases given 3 cycles treatment with GnRH-a.12 patients who required recanalization due to fallopian tube ligation served as the control group.Collected the peripheral blood at different time,sush as preoperation,1 month after operation and 3 months after operation/or after 3 cycles of GnRH-a treatment.Flow cytometry was used to detect the ratio of Th17/Treg cells in the peripheral blood,ELISA detected the expression levels of IL-6,IL-10,IL-17 A and TGF-β,which were related with Th17 and Treg cells,in serum and peritoneal fluid.A proper amount of peritoneal fluid was collected firstly,and the cyst wall of the Ovarian endometriotic cyst was obtained about 1cm×1cm in size during the surgery.Among the 40 patients,22 cases were treated with hysteroscopy at the same time,and the eutopic endometrium tissue was taken.Peritoneal fluid was firstly collected during the operation in the control group,of which 10 cases required hysteroscopy to obtain the eutopic endometrial tissue at the same time.Immunohistochemistry was used to detect the expression of RORγt and Foxp3 in ectopic and eutopic endometrium tissues.Results:(1)Compared with the control group preoperative:(1)the proportion of Th17 cells which elevated(21.47±8.05)% in the peripheral blood of the EMS group preoperative increased significantly,Treg cells(5.57±2.71)% decreased significantly,and the ratio of Th17/Treg(2.94±1.14)increased significantly,P<0.05;(2)The levels of IL-6,IL-10 and IL-17 A [respectively(122.84±39.98),(147.46±48.89)and(6.25± 3.87)ng/L] in serum were increased significantly,and TGF-β(32.87±12.27)μg/L decreased significantly,P<0.05;(3)The levels of IL-6,IL-10 and IL-17 A in the peritoneal fluid of the EMS group [Respectively(250.57±91.77),(395.28±150.35)and(42.21±38.29)ng/L] increased significantly,and TGF-β(2.05±0.40)μg/L increased significantly,P<0.05.(2)Compared with the normal endometrium of the control group:(1)The protein expression of RORγt in the eutopic and ectopic endometrium of the EMS group increased significantly,P<0.05;(2)The Foxp3 in the ectopic endometrium increased significantly,P<0.05,there was no significant difference in the protein expression of Foxp3 in the eutopic endometrium.(3)Compared with the EMS group preoperative:(1)The proportion of Th17 cells(11.78±2.86)% decreased significantly in the peripheral blood of the EMS group 1 month after the operation,Treg cells(7.51±2.37)% increased significantly,the ratio of Th17/Treg(1.79±0.41)decreased significantly,P<0.05;(2)The levels of IL-10 and IL-17A[respectively(69.29± 29.72)and(3.88±2.40)ng/L] reduced significantly in serum,TGF-βlevel(47.97±13.07)μg/L increased significantly,P<0.05,but IL-6(102.93±25.02)ng/L decreased,P>0.05.(4)(1)Compared with the EMS group 1 month after operation,the proportion of Th17 and Treg cells and the ratio of Th17/Treg changed slightly in peripheral blood of the surgery-only treatment group 3 months after the operation and the GnRH-a treatment group,P>0.05;The levels of IL-6,IL-10 and TGF-β continued to approach the control group,P>0.05,while the level of IL-17 A increased significantly,and P<0.05;(2)There was no significant difference between the surgery-only treatment group and the GnRH-a group 3 months after the operation.(5)Compared with preoperative,the proportion of Th17 and Treg cells,the ratio of the two,and the levels of related cytokines in the control group made no significant sence at 1 month and 3 months after operation.Conclusion:(1)Th17/Treg cells and cytokines related are imbalanced in patients with endometriosis,which are closely related to the immune pathogenesis of EMS;(2)Laparoscopic surgery can partially restore the immune imbalance in patients with endometriosis;(3)GnRH-a treatment has no significant effect on the immune balance of patients with endometriosis;(4)Overexpression of RORγt and Foxp3 may also be involved in the pathogenesis of endometriosis.