Expressions Of VEGF, TNF-α, IL-18and Their Relationships In The Endometrium Of Patients With Intrauterine Adhesions

Intrauterine adhesions mean that a variety of reasons such as trauma, severe infections may damage the stratum basale of the endometriums, this caused adhesions formed in the uterus which could make uterine cavity change its shape or die out. Patients suffering from IUA show menstrual disorders(amenorrhea,hypomenorrhea,et al), sterility, miscarriage, premature,pelvic pain and so on. Most scholars think that the pathogenesis of IUA may be mainly concerned with trauma and infections. In recent years, IUA prevalence rate rises year by year. The morbidity of IUA in women who accept induced abortion is37.6%.The postoperative recurrence of IUA is high, especially in the patients with severe adhesions. The pathogenesis of IUA is unclear. The researches at present mainly focus on the relationship between profibrogenic cytokines and IUA.However, the function of infection in the the process of intrauterine adhesions, return existence controversy..There will be vasculogenesis and inflammatory mediators when the endometriums suffer from trauma and infections. Vascular endothelial growth factor (VEGF) is an endothelial-specific multifunctional cytokine that generated by tissue cells, could increase venule and small vessel permeability, induce vascular endothelial mitosis and proliferation and accumulation of abundant calcium precipitates in cytoplasm and angiogenesis. VEGF plays a very important part in the formation of chronic inflammation.VEGF binded its receptors could induce the growth, differentiation, proliferation and transference of vascular endothelial cells, promote neovascularization, and support the fibrosis of endometrium. VEGF has high affinity to fibrinogen and fibrin monomer.The accumulation of fibrin monomer could induce fibroblast proliferation, and then increase synthesis of ECM. TNF-α is a kind of inflammatory mediator and immune factor, could enhance the activities of neutrophils and eosinophils; induce the formulation of inflammatory mediator (IL-6, IL-8,PG and so on) in cells of blood vessels, then a local inflammation come up which resulted in the production of the inflammatory mediator in cells of blood vessels and leukocytes; In addition, it is a kind of profibrogenic cytokines and could increase proliferation of fibroblast. IL-18is a kind of pro-inflammatory cytokines,which plays an important role in infection and tumor, it could induce the release of cytokines (IFN-γ; TNF-α) which is generated by immune cells (Thl cells,NK cells). IL-18could induce the release of TNF-α,and then TNF-α could induce the release of IL-18,it form a positive feedback effect.ObjectiveTo study the expression of VEGF,TNF-a,IL-18in the endometrium tissues of patients suffering from IUA and uterus septus. And to investigate their significances in the development of intrauterine adhesions by immunohistochemistry.Materials and Methods1Object of study60patients who were hospitalized in the Maternal and Child Care Service Centre of Henan because of intrauterine adhesions from October2010to March2011were chosen, including32patients in moderate IUA group (aging27.53±3.10)and28patients in severe IUA group (aging27.75±4.18) according to the MARCH classification system.Among these patients,8patients had amenorrhea,30patients had spanomenorrhea and22patients had secondary infertility,30patients aging (27.43±3.80) hospitalized in the Maternal and Child Care Service Centre of Henan because of uterus septus within the same period were selected to be the control group.2Samples CollectionWash endometrium tissues from90patients with normal saline, immerse the tissues into4%paraformaldehyde solution to last for24hours, then make them paraffin-embedded routinely.An incisional biopsy was done with4u m thickness for immunohistochemistry.3MethodsThe VEGF,TNF-α and IL-18in endometrium tissues were detected with the immunohistochemistry S-P methods with PBS as blank control replace first antibodies. The experiments were conducted strictly compliance with the requirements of the reagents. Upon observation under the microscope, immediately terminated reactions after Color reaction showed colorfully. Observe carefully on all the slices and filming under the microscope.Collecting images with matched image acquisition system and using Imaging system to analyse each slice with the integral optical density as measure. Selecting casually five high power fields (10X40), average IOD value of the positive section which was used as the measurement value of each slice was measured.Results1The expression of VEGF in the endometriumThe expression of VEGF appeared in three groups of endometrium, the positive cells showed yellow-brownish. Their expressions of endochylema of the endometrium glandular epithelium are higher than that of mesenchyma.The expression of VEGF in the endometrium of severe adhesion group was 155.51±5.24, the moderate adhesion group was133.23±5.57, the control group was132.08±4.67. The severe adhesion group was higher than that of moderate adhesion and control group (P<0.05), and the expression of VEGF in moderate adhesion group was higher than that in the control group (P>0.05).2The expression of TNF-α in the endometriumThe expression of TNF-α appeared in three groups of endometrium, the positive cells showed yellow-brownish. Their expressions of endochylema of the endometrium glandular epithelium are higher than that of mesenchyma. The expression of TNF-α in the endometrium of moderate adhesion group was139.38±5.53, the severe adhesion group was147.71±5.05, the control group was120.49±5.19. The severe adhesion group was higher than that of moderate adhesion and control group (P<0.05), and the expression of TNF-a in moderate adhesion group was higher than that in the control group (P<0.05).3The expression of IL-18in the endometriumThe expression of IL-18appeared in three groups of endometrium, the positive cells showed yellow-brownish.Their expressions of endochylema of the endometrium glandular epithelium are higher than that of mesenchyma. The expression of IL-18in the endometrium of moderate adhesion group was132.41±4.86, the severe adhesion group was159.50±3.53, the control group was105.91±4.66. The endometrium of severe adhesion group was higher than that of moderate adhesion and control group (P<0.05), and the expression of IL-18in moderate adhesion group was higher than that in the control group (P<0.05).4The relevance of VEGF, TNF-α and IL-18in moderate and severe adhesion groupsIn order to definite the role of VEGF, TNF-α and IL-18in the pathogenesis of IUA, the relevance of them expressed in moderate and severe adhesion groups was analysed. VEGF was correlated positively with TNF-α and IL-18in the endometrium of moderate adhesion group (P<0.05).In the endometrium of severe adhesion group, VEGF was correlated positively with TNF-a and IL-18(P<0.05).Conclusions1. VEGF, TNF-α and IL-18may play important roles in the development of IUA.2. The expression of VEGF, TNF-a and IL-18may be concerned with the severity of IUA.