Research The Role About NF-κB In Pathogenesis Of Endometriosis

Backgroud Endometriosis is a kind of common and incurable gynaecology disease, the incidence of it about 15％in reproductive female, and now it has obviously upgrade tendency. It causes menorrhalgia, hypogastralgia, algopareunia, et al. It affects the healthy and quality of life severely, and becomes one of main causes of infertility. The pathological changes are extensive and diversity, the biological behavior is malignant, such as infiltrative growth,implantation metastasis widely and recure easily, so it becomes one of troublesome gynaecology diseases. The pathogenesis of it is still unkown, Up to now, the hypothesis of endometrium exfoliative fragments transplantation which Sampson put forward in 1925 has been accepted by most scholars. It mainly includes three stages: adhesion, invation and angiogenesis, involve in numerous cytokines,enzymes,growth factors, and heredity,dyscrinism,immune function abnormity. Recently, people have thought highly of the eutopie endometrium's feature and role.Nuclear faetor-kappaB(NF-κB), which resides in eukaryotic cells' kytoplasm with unreactive trait, a key nuclear transcription factor actived by physiology or pathology stimulus, plays an important role in accommodation of cytokine, cytomembrane receptor,membrane adhesion molecule,acute-phase proteins(APP),growth factor,transcription factor, et al.So NF-κB is related to the cells' growth,differentiation,inflammatory reaction,immune response,tumor growth, and so on.Recently, NF-κB has become a hot spot, and has some correlation with liver cancer,prostatic carcinoma,gastric cancer,bronchovesicular cancer,rectal cancer,breast cancer,cervix cancer and neurospongioma, et al. But there is little research between NF-κB and endometriosis, abroad scholars found that TNF-αand IL-1βcan stimulus monocyte to cause RANTES increased through NF-κB signal channel, which causing endometriosis. So the purpose of us is approaching the role of NF-κB in pathogenesis of endometrioisis.Objective 1. We detect NF-κBp65,VEGF,ICAM-1,MMP-9 and Bcl-2 in eutopic and ectopic endometrium by immunohistochemical method, and appoach the associativity between NF-κBp65 and VEGF,ICAM-1,MMP-9,Bcl-2.2. In vitro culture endometrial cells,eutopic and ectopic endometrial cells, we add with LPS and LPS+PDTC,(1)detecting the three groups endometrial cells' NF-κB p65 protein by Western Blot, we can learn the influence of LPS and LPS+PDTC to the eutopic and ectopic endometrial cells. (2) Dectecting the three groups of culture endometrial cells' fluid by ELISA, we can learn the concemtration of IL-1β,TNF-αand the influence of LPS and LPS+PDTC to the three groups endometrial cells. From the results we got maybe we can obtain some hinds about the pathogenesis of endometriosis.3. We establish the rat endometriosis model, using NF-κB inhibitor NF-κB antisense oligonucleotides to cure the models, then observing the influence to the modeling rate,ectopic focus and TNF-αmRNA. From the results we got, we may learn the role of NF-κB in endometriosis.Method and Result 1. The expression of NF-κB in endometriosis Collecting the endometriosis patients' eutopic and ectopic endometdum, the normal endometrium as control, we dectect the expression of NF-κB,VEGF,ICAM-1,Bcl-2,MMP-9 in those specimen by immunohistochemistry. The results we got are: (1) They express in the endothelial cells,the matrix and blood vessel endothelium have a litter. Comparing with the normal endometrium, the eutopic and ectopic endometrium have obviously differences. (2) The expression of NF-κB in eutopic and ectopic endometrium has no cyclic change. (3) The expression of NF-κB has some relation with the stage. The expression of NT-κB in stageⅡis strongest than other stages in eutopic endometrium, but in ectopic endometrium, the expression of NF-κB in stageⅠandⅡare stronger than stageⅢandⅣ. (4) There are positive correlation between NF-κB and Bcl-2,VEGF,ICAM-1 and MMP-9. The correlation coefficient are 0.77, 0.5, 0.63 and 0.57.2. In vitro cultured endometriosis cells, the influence of LPS or LPS+PDTC to NF-κB protein and IL-1β,TNF-αconcentration in three groups endometrial cells. (1) In vitro cultured endometial cells,eutopic and ectopic endometrial cells, we add with LPS or LPS+ PDTC, then detect the NF-κB protein in three groups by Western Blot, and compare the NF-κB protein concentration and the changes after intervention. (2) We collect the three groups of culture fluid, then dectect the concentrateon of IL-1βand TNF-α, compare the difference of three groups and the changes after intervenetion. The results we got are: (1) The concentration of NF-κBp65 protein in eutopic and ectopic endometrial cells is more than normal endometrial cells. LPS can stimulate more NF-κB protein, PDTC has repressive role. (2) The concentration of IL-1βand TNF-αin eutopic and ectopic endometrial cells' culture fluid is higher than normal endometrial cells. LPS can increase the the concentration of IL-1βand TNF-αin eutopic and ectopic endometrial cells' culture fluid, but PDTC has opposite role.3. We establish the SD rat endometriosis medel and inject the NF-κB ASODN from the vena caudalis, then observe the modeling rate,the changes of ectopic focus. Then we collect the ectopic focus and detect the TNF-αmRNA by RT-PCR. The results show: (1) The modeling rate lower and the ectopic focus shrink than controls after NF-κB ASODN injection, but in the two control groups, the focus becomes bigger with the time extend, the 8 weeks is bigger than the 4 weeks. (2) The expression of TNF-αmRNA is weaker than the two controls after NF-κB ASODN injection.Conclusion (1) NF-κB has abnormal higher expression in eutopic and ectopic endometrium of endometriosis patients, and there is litter expression in normal endometrium. These results demonstrate that NF-κB has some relation with endometriosis.(2) The expression of NF-κB has no cyclic change and play the role mainly in the early and mid-stages.(3) There are positive correlation between NF-κB and VEGF,ICAM-1,Bcl-2,MMP-9, which show NF-κB may play role in endometriosis by up-regulate the expression of VEGF,ICAM-1,Bcl-2,MMP-9.(4) The protein expression of NF-κB is higher in eutopic and ectopic endometrial cells. LPS can raise up the protein of NF-κB in eutopic and ectopic endometrial cells, and increase the concentration of IL-1βand TNF-α, then PDTC can restrain these role, which explains NF-κB as a key regulatory factor, maybe it acts as a trigger point role in the pathogenesis of endometriosis, controls the IL-1β,TNF-a and other factors.(5)Using the NF-κB ASODN to cure the rat endometriosis models, we fond that the modeling rate is lower and ectopic focus shrinke obviously, the expression of TNF-αmRNA cut down remarkably. All illustrate that NF-κB ASODN is effect to the rat endometriosis model, further verificate there is intimate relation between NF-κB and endometriosis.