Expression And Significance Of MEK/ERK And NF-κB In Endometriosis Rats

Objective 1.Establish a rat model of endometriosis,To investigate the expression and correlation of MEK、ERK and NF-κB in rat endometrial tissue.2.To investigate the changes of clinical characteristics and cell proliferation and invasion activity of endometriosis rats after MEK/ERK and NF-κB related inhibitors.Methods 1.Twenty-five rats of SPF grade SD were selected,Rat model of endometriosis was established by autologous transplantation(15 rats),in which sham operation group was used as control group(10 rats);The growth of ectopic lesions was observed by second laparotomy after 4 weeks,HE dyeing identification model is successful,Immunohistochemistry and Western blot were used to detecte the expression of MEK、ERK and NF-κB proteins in the eutopic endometrium group and the ectopic endometrium group and control group,Pearson correlation analysis of correlation between each factors.2.Fourty SPF grade female non-pregnant SD rats were selected as subjects,eight rats were randomly selected as control group and the remaining 32 rats EMs established.the second laparotomy after 4 weeks of modeling,24 rats was successfully established the EMs model,and were randomly divided into4 group:model group,DMSO group(drug solvent),MEK/ERK inhibitor group(U0126 group)and NF-κB inhibitor group(BAY11-7082 group),with 6 rats in each group.Among them,blank control group and model group:The same volume of normal saline was given to the stomach three times a week;MEK group and NF-κB inhibitor group:20mg/kg intraperitoneally injected three times a week,after four weeks of continuous administration,Unified execution of rats,Measurement of volume changes in ectopic lesions;Histopathological changes of ectopic lesions in rats were observed by HE staining;The expression of MEK、ERK and NF-κB in intimal tissue and the proliferation(PCNA)and invasiveness(MMP9)of local lesions were detected by immunohistochemistry and Western blot.Results 1.MEK and ERK were mainly localized in the cytoplasm of endometrial epithelial cells of EMs rats,NF-κB was mainly localized in endometrial glands and nuclei of glandular epithelium of EMs rats.2.The expression of MEK、ERK and NF-κB protein in rat endometrium:immunohistochemistry and Western-blot results showed that the expression of MEK、ERK and NF-κB protein in the eutopic endometrium group and the ectopic endometrium group was significantly higher than that in the control group,moreover The expression of EMs in ectopic endometrium group was higher than that in eutopic endometrium group(P<0.05).3.The expression correlation of MEK、ERK and NF-κB in EMs rat endometrium:the expression of protein was positively correlated in EMs rat eutopic endometrium group and the ectopic endometrium group.EMs eutopic endometrial group(r MEK/ERK=0.634,r MEK/NF-κB=0.721,r ERK/NF-κB=0.709,(P<0.05)),EMs ectopic endometriosis group(r MEK/ERK=0.661,r MEK/NF-κB=0.662,r ERK/NF-κB=0.657,(P<0.05)).4.The volume changes of ectopic lesions between MEK/ERK and NF-κB inhibitors:The volume of ectopic lesions in MEK/ERK inhibitor group and NF-κB inhibitor group(U0126 and BAY11-7082)were decreased significantly compared with model group(P<0.05).5.Histopathologic changes of intima after drug intervention with MEK/ERK and NF-κB inhibitors:HE staining showed that:compared with the blank control group,the EMs model group showed multiple endometrial ectopic gland cysts and interstitial reactions of different sizes,with multiple inflammatory cell infiltration,hemorrhage and neovascularization around the glands;compared with the EMs model group,the MEK/ERK inhibitor group(group U0126)showed a decrease in endometrial glands,a smaller glandular cavity,a thinner glandular epithelium,a reduced interstitial response,and a decrease in inflammatory cells.NF-κB inhibitor group(BAY11-7082 group)showed that the residual endometrial glands was decreased and The gland cavity decreases and the interstitial inflammatory response significant decrease,with only a few inflammatory cell infiltration and a few hemosiderin macrophages.6.The changes of MEK/ERK and NF-κB protein expression in eutopic and ectopic endometrial tissue after drug intervention with MEK/ERK and NF-κB inhibitor:Immunohisto-chemical and Western-blot results showed that compared with the control group,The expression levels of MEK/ERK and NF-κB proteins in the model group and the DMSO group were significantly increased,Differences between groups were statistically significant(P<0.05);The expression of MEK/ERK and NF-κB proteins in the group of MEK/ERK inhibitors(group U0126)was significantly decreased in the in eutopic and ectopic endometrial tissues,Differences between groups were statistically significant(P<0.05);The expression of NF-κB proteins in NF-κB inhibitor group(BAY11-7082 group)was significantly decreased in both in eutopic and ectopic endometrial tissues,Differences between groups were statistically significant(P<0.05),but MEK and ERK expression levels were not statistically significant(P>0.05)。7.Invasive changes of tissue proliferation in eutopic and ectopic endometrium after MEK/ERK and NF-κB inhibitor intervention:Immunohistochem-ical and Western-blot results showed that compared with the control group,The expression levels of PCNA and MMP9 proteins in the model group and the DMSO group were significantly increased,Differences between groups were statistically significant(P<0.05);The expression of PCNA and MMP9 in MEK/ERK inhibitor group(U0126 group)and NF-κB inhibitor group(BAY11-7082 group)eutopic and ectopic endometrial tissue was significantly decreased,Differences between groups were statistically significant(P<0.05).Conclusion 1.The expression of MEK、ERK and NF-κB was increased and positively correlated in the eutopic endometrium and ectopic endometrium of EMs rats;2.MEK/ERK and NF-κB inhibitor drugs can significantly inhibit the growth of ectopic lesions in EMs rats and reduce the hyperplasia of glands and interstitial inflammatory response in ectopic lesions,The change of growth activity of ectopic lesion tissue EMs the expression of proliferative invasive protein is closely related to the activity of MAPK/ERK and NF-κB pathway,and then the purpose of reducing the EMs ectopic lesion in the treatment of endometriosis.