Expression And Significance Of TLR4, MyD88and Ap-1in Endometriosis

Endometriosis（endometriosis,EMs）. It is an inflammatory and estrogen-de-pendent disease defined by the presence of viable endometrial tissue outside the uterine cavit, mainly on the pelvic peritoneum, but also on the ovaries and in the rectovaginal septum. EMs has a variety of clinic characters and the main symptoms are dysmenorrhea, infertility, menstrual flow abnormalities. Histology of EMs is benign, but there is proliferation, invasion, metastasis and recurrence of malignant behavior.Estimates show that endometriosis affects10-15%of women and the rate is increasing year by year. Currently, the treatment of the disease is still hormone-based drugs and surgical, but its effect is not ideal, seriously affecting the quality of life and physical and mental health of female patients.The causes of endometriosis are multifactorial, complex and vary between individuals. Currently, The planting theory was most widely accepted. Sampson first proposed The blood reflux planting theory in1921, the origin of ectopic endometrial tissue is a combined effect of retrograde menstruation and associated implantation of endometrial fragments at an ectopic site. However, about90%of women in the retrograde menstruation, but it is unclear that why clinical endometriosis develops in only15%to20%of women during their reproductive life. Some scholars have pointed immune factors, genetic factors, inflammatory factors in the pathogenesis of EMs. Jinghe Lang presented for the first time in the international "the eutopic endometrium determinism" is a new milestone in the study of the pathogenesis of EMs. It is considered to be that the differences of Eutopic endometrium between endometriosis and non-endometriosis involving genetic,gene expression and, adhesion, invasion, angiogenesis ability to significantly enhance.These make it easier to form endometriosis. Studies showed that the growth and progres-sion of endometriosis continue even in ovariectomized animals. This indicates that besides ovarian steroid hormones, the growth of endometriosis can be regulated by the innate immune system in the pelvic environment.Toll-like receptors （Toll-like receptors, TLRs） are a recently discovered innate immune receptor and involved in the innate immune as key role. There are two significant biological function:The one hand, to promote the synthesis and release of cytokines, triggering the inflammatory response; The other is to promote the maturation of antigen-presenting cells, inducing the body acquired immune response. TLR4involved in cell signal transduction pathway includes MyD88-dependent and non-dependent. MyD88-dependent signaling pathway is both divided into NF-κB and AP-1two-ways.Activation of TLR4by of TLR4/MyD88/NF-κB and TLR4/MyD88/AP-1two signaling pathways induce some columns inflammatory mediators, Including cytokines, inflammatory cytokines and chemokines, resulting in a strong inflammatory response. ObjectiveTo investigate the expression and clinical significance of the Toll-like receptor4（TLR4） mediated pathway via myeloid differentiation factor88（MyD88） and activator protein-1（AP-1） in eutopic and ectopic endometrium of endometriosis （EMs）Materials and Methods1Object of study 45cases of patients with EMs were selected（EMs group,15cases at I and Ⅱ,30cases at III and IV）,and45cases of ectopic endometrium （ectopic endometrium group）,and30cases of eutopic endometrium （eutopic endometrium group） were obtained.During the same period30pationts with septate uterus undergoing hysteroscopic surgery were selected as control group and their uterine endometrium were obtained.The expression of the TLR4and MyD88were evaluated by Semi-quantitative RT-PCR and immunohistochemical technique （SP） in above groups.2MethodsThe expression of the TLR4、MyD88and AP-1were evaluated by Reverse T ranscription-PCR and immunohistochemical technique （SP） in ectopic endometrium、 eutopic endometrium、normal uterine endometrium tissues.3Statistical methods:SPSS17.0was performed to analysis the study data. All the data was express as mean±standard deviation（x±s）.The differences in means were analyzed by t-test（two groups） or ANOVA（more than two groups） and LSD-t method in pairwise comparisons between groups.The relationship of two variables was analyzed by correlation analysis.Differences with P＜0.05were regarded as statistically.Results1. The expression of TLR4,MyD88and AP-1mRNA and protein in ectopic endometrium group,eutopic endometrium group,and control group were statistically significant（P＜0.05）,and those in ectopic endometrium group（mRNA:0.156±0.017,0.272±0.023,0.243±0.026）、（protein:60.765±15.550,59.628±8.016,43.967±3.836） and eutopic endometrium groups （mRNA:0.139±0.015,0.243±0.025,0.214±0.024）、（protein:42.196±12.388,52.372±7.160,35.200±3.587）were significantly higher than in control group（mRNA:0.124±0.014,0.221±0.028,0.196±0.035）、（protein:30.786±11.121,46.772±5.020,31.437±3.221）（P＜0.05）,and those ectopic endometrium group was significantly higher than eutopic endometrium groups（P＜0.05）. 2. No significant difference of the expression of TLR4,MyD88,AP-1mRNA and protein were found between early stage（mRNA:0.154±0.019,0.269±0.028,0.231±0.027）、（protein:57.793±16.380,59.047±8.338,41.376±4.151） and advanced stage （mRNA:0.156±0.016,0.274±0.021,0.250±0.024）、（protein:62.251±15.182,59.919±7.978,45.262±2.967） of endometriosis（all P＞0.05）.3. Positive correlation ship was found between the expression of the TLR4and MyD88,AP-l,respectively,in both of ectopic endometrium group（rmRNA:0.812,0.719, P＜0.05; rprotein:0.639,0.648, P＜0.05）and eutopic endometrium group (rmRNA:0.594,0.679, P＜0.05; r_protein:0.560,0.575, P＜0.05).In the control group,TLR4and MyD88,AP-1, respectively, there was no significant correlation （rmRNA:0.334,0.097, P＞0.05; rprotein:0.358,0.170, P＞0.05）.Conclusions1. TLR4/MyD88/AP-1signal transduction pathway may play an important role in the pathogenesis of endometriosis.2. TLR4/MyD88/AP-1signal transduction pathway may involved in the early onset of endometriosis, does not affect the progression of the disease.