Expression Of NF-κB And MIF In EPs Among Women Of Reproductive Age

Endometrial polyps (EPs) is a common gynecological endometrial lesions,which can cause menstrual increased,prolonged menstrual,bleeding between periods,and even the possibility of malignancy,but some patients don’t have any clinical symptoms.In recent years,with the widespread use of transvaginal ultrasound and hysteroscopy,the detetion rate of EPs has been increased year by year.For the women of childbearing age, the breadth of its incidence seriously affects their quality of life and fertility requirements.Therefore,it’s particularly important to study pathogenesis and treatment of EPs.During clinical work,transvaginal ultrasound can show EPs by thickening of the endometrium,uneven endometrial echo,focally enhanced clump echo,and so on.Punctate or short strip bloodstream signal can be displayed at the bottom of minority polyps by transvaginal ultrasound.EPs is smooth pedunculated soft vegetation by hysteroscope,which is mostly willow-like or tongue-like,and single or multiple.EPs is composed of focal fibrotic interstitial and glands at least,and its interstitial owns thick-walled blood vessels.Most scholars think that the pathogenesis of EPs finally due to reactive hyperplasia of the endometrium and angiogenesis. Consequently,reactive hyperplasia of the endometrium and angiogenesis is an significant factor in the progression and development of endometrium polyps. Nuclear factor kappaB (NF-κB) is widely present in eukaryotes,which is transcription factors of multidirectional regulation.It’s the common pathway and sites of action which contain a variety of cytokines and inflammatory mediators gene transcription and the cascade process and sites of action. Its peptide p50and p65subunits form homologous or heterologous dimer. NF-κB plays a central role in different cellular processes,such as inflammation,immune response,differentiation, proliferation,apoptosis and tumorigenesis.Recently,study find that NF-κB relates to the development of a variety of diseases,for example,endometriosis,endometrial cancer,nasal polyps.However, the role of NF-κB in EPs is resported to the least extent.Incidence of EPs involves endometrial reactive hyperplasia,angiogenesis and the participation of a number of cytokines and growth factors.Abnormal activation of NF-κB may raise relevant cytokines.Macrophage migration inhibitory factor (MIF) is a crucial pro-inflammatory cytokines,with a variety of biological functions,which can inhibit macrophage walk,promote macrophages secrete a lot of inflammatory cytokines to paticipate in the regulation of the inflammatory response.It can also stimulate cell proliferation,when it’s promoted in a variety of growth factor and cytokines, and it can induct the expression of genes which can promote angiogenesis.Studies have shown that MIF is a downstream gene of NF-KB,subjected to the regulation of the NF-κB.However,it’s unclear whether NF-κB and MIF play an impotant role in the pathogenesis of EPs.ObjectiveReverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemical method were used to detect the expression of NF-κB and MIF in EPs and the endometrium around EPs and normal endometrium of septate uterus in this experiment,we evaluated the relationship between NF-κB and MIF with EPs among women of reproductive age and the expression correlation between NF-κB and MIF in EPs.in order to provide theory basis for the clinical diagnoses and prognosis of EPs and to find a new target for treatments of EPs. Materials and Methods1Object of studyThe samples were collected from40patients with EPs (40cases of EPs and the endometrium around EPs) and22patients with septate uterus (normal endometrium) during the hysteroscope operation from September in2011to February in2012. All the tissue have maken a definite diagnosis.The distance between EPs and the endometrium around EPs is at least0.5cm.EPs and the endometrium around EPs are the experimental groups (aging27.975±3.939),and normal endometrium is the control group (aging26.636±3.672).Two group ages were no significant difference (P>0.05).The operations were done in third to seventh days after menstruation.All the patients who hadn’t own other malignant diseases and serious complications didn’t use any antibiltics or hormone therapy or IUD,and didn’t accept uterine surgery during three months before surgery.All the patients hadn’t own other genital disease,for example, adenomyosis and uterine myoma and so on.2Samples CollectionWe obtained specimens under sterile conditions,some tissue was saved in liquid nitrogen (temperature:-80℃),the other was fixed in10%formalin solution embedded in paraffin,and sliced,which was used finally after pathologically confirmed.3MethodsNF-κBmRNA and MIFmRNA expression was detected by reverse transcriptase polymerase chain reaction(RT-PCR) and expression of NF-κB protein and MIF protein by immunohistochemical SP method.We used two methods respectively from the gene to protein levels to resolve the different expression of NF-κB and MIF in the three group (EPs, the endometrium around EPs and normal endometrium) and analyse the correlation between NF-κB and MIF.Finally,in order to explore the role that both can play in the development of EPs among women of reproductive age. 4Statistical MethodsStatistical anylysis was performed using the SPSS17.0statistical software,datas were analyzed by single-factor analysis of variance test,when datas conform to heterogeneity of wariance,wilcoxon test was used.their relationship of NF-κB and MIF were analysised by Pearson rank correlation test, α=0.05acted as level of significance,comparison of test level correction between two groups=0.0167.Results1The expression of NF-κB protein in different tissueThe expression product of NF-κBp65protein showed brown stain,the average integral opartial density value of NF-κBp65protein was91.357±7.470in the EPs group,the endometriun around EPs group was68.258±6.290,the control group was34.247±3.977. The staining of NF-κB protein was found in the three groups.lt was expressed most in EPs.The expression of it took second place in the endometrium around EPs,pairwise comoarisons between three groups were statistically significant differences(P<0.0167).2The expression of MIF protein in different tissueThe expression product of MIF protein showed yellow-brownish,the average integral opartial density value of MIF protein was182.895±13.664in the EPs group,the endometriun around EPs group was143.128±12.063,the control group was70.595±6.656.The expression of MIF protein in the EPs group was higher than that of the endometriun around EPs group and control group(P<0.0167),and the expression of the endometriun around EPs group was higher than that of in the control group(P<0.0167).3The expression of NF-κBmRNA in different tissueThe expression of NF-κBmRNA appeared in the three groups.According to the gray value,the expression of NF-κBmRNA was0.683±0.025in the EPs group,the endometriun around EPs group was0.207±0.013,the control group was0.092±0.006.The expression levels of NF-KBmRNA in EPs were significantly higher than the endometrium around EPs and normal endometrial tissues(P<0.0167),and the expression levels of NF-κBmRNA were higher in the endometrium around EPs than the normal endometrial tissues in the same period(P<0.0167).4The expression of MIFmRNA in different tissueThe expression of MIFmRNA were found in the three groups.According to the gray value,the expression of MIFmRNA was0.845±0.052in the EPs group,the endometriun around EPs group was0.322±0.023,the control group was0.123±0.007.The expression levels of MIFmRNA in EPs group were significantly higher than the endometrium around EPs group and normal endometrial tissues group(P<0.0167),and the expression levels of MIFmRNA were higher in the endometrium around EPs group than the normal endometrial tissues group in the same period(P<0.0167).5The relevance of NF-κB and MIF in EPs groupThe relevance of NF-κB and MIF expressed in EPs group was analysed to definite the effect of them in the pathogenesis of EPs. NF-κB was correlated positively with MIF at the protein level(r=0.689,P<0.05).NF-κB and MIF had a positively correlation at the gene leve(r=0.408,P<0.05).Conclusion1. The high expression of NF-κB and MIF suggests that it may be involved in the occurrence of EPs during the women of childbearing age.2. NF-κB and MIF may synergistically promote the development and evolution of endometrial polyps.