Expressions And Significance Of PDGF, TGF-β And TIMP-1 In Endometrium Tissue And Serum Of The Women With Intrauterine Adhesions

The intrauterine adhesions (IUA) refers to the mutual adhesion between the cervical canal and/or uterine cavity due to the reasons including intrauterine surgery, radiation, infection, tuberculosis, etc with the clinical manifestation including periodic abdominalgia, spanomenorrhea or amenorrhea, secondary infertility, abnormal pregnancy (abortion, premature delivery and utero fetal death, etc) and abnormal placenta (placenta previa, placenta accrete and placenta accrete, etc). With the increasing frequency of induced abortion and various intrauterine operations, IUA appears a rising tendency year by year in recent years. It is reported that about 1.7% secondary amenorrhea patients and 40% infertility patients have intrauterine adhesions to different extents. However, its pathogeny and pathogenesis are not completely known and become the difficult and hot topics in gynaecology research field, therefore, the domestic and foreign scholars have conducted plenty of clinical observation and experiment research and propose the following theories regarding its pathogeny and pathogenesis:1) trauma; 2) infection; 3) nerve reflex doctrine; 4) endometrium fibrosis theory; 5) low estrogen state.Recently, the endometrium fibrosis theory gains the most attention and becomes the hot topic and difficult point of the research. PDGF is the key cell factor to control the embryo's growth and development, differentiation, cell growth and many other diseases including cancer. It is reported that PDGF has close relation with the occurrence and development of various organ fibrosis; TGF-βis the powerful cell proliferation regulator that promotes EDM's composition and restrains its degradation in the fibrosis process, plays roles in each link to stimulate the composition and sedimentation of various types of ECM, reducing matrix-degrading enzymes while increasing the quantity of stromatin receptors and increases the composition of the degrading enzyme inhibitors thus to promote the occurrence of fibrosis. TIMP-Ⅰis the main inhibitor for a group of matrix metallo proteinases (MMP), almost irreversibly restraining MMP's activation and preventing ECM's degradation to promote ECM's accumulation outside the cell and the fibrosis formulation.ObjectiveThe research adopts IHCA and ELISA to make respective inspection for the representation of PDGF, TGF-βand TIMP-1 in the endometrium tissues and serum of the intrauterine adhesions patients and non-intrauterine adhesions patients to discuss the roles that PDGF, TGF-βand TIMP-1 play in the occurrence and development of intrauterine adhesions to provide theory basis for the treatment and prevention of intrauterine adhesions.Materials and Methods1 Objects60 IUA patients hospitalized in the Third Affiliated Hospital of Zhengzhou University because of IUA from September 2009 to December 2009 are chosen to be the research group, including 30 cases in mild to moderate adhesion group aging (29.57±3.53) and 30 cases in severe adhesion group aging (29.82±3.85); regarding the chief complaint, ten cases of amenorrhea,33 cases of spanomenorrhea and 17 cases of secondary infertility.30 TCRS patients aging (28.77±4.60) hospitalized in the Third Affiliated Hospital of Zhengzhou University because of uterus septus within the same period are chosen to be the comparison group.Admission standard:Intrauterine Adhesions Group:having diagnosed to be IUA patients upon hysteroscopy and divided into mild to moderate adhesion group and severe adhesion group according to March classification method. Uterus Septus Group:having diagnosed to be uterus septus patients upon hysteroscopy. Exclusion standard:age>40; weight>100 kg; having taken hormone medicine or placed IUD within six months before the operation; demonstrating to have other intrauterine diseases (such as myoma, polyp, etc) upon hysteroscopy; having other serious systematic diseases (blood coagulation disorders, immunologic diseases and serious cardiovascular, liver and kidney diseases and tumour); in the acute inflammation reaction stage; or with serious allergy history and anaphylaxis. All the research objects have signed the information consent form.2 Sample Collection1) Endometrium tissues:respectively fetch the endometrium tissues from IUA patients and the comparison group, put the endometrium tissues into 10 formaldehyde solution for 24 hours and then embed with paraffin. After HE dye each sample, observe the histopathologic morphology under the microscope and select the tissue samples.2) Respectively fetch 3 mL peripheral venous blood in empty stomach from IUA patients and the comparison group under the sterile operation; get the supernate after centrifugation, then number in groups and place in -80℃refrigerator for frozen storage to be tested in batches.3 Methods1) The endometrium tissues adopts the immunohistochemistry S-P method to be dyed in strict accordance with the instruction method on the kit. Upon careful observation under the microscope, it is the positive cell if the endochylema or the cell membrane of the endometrium tissues show yellow-brownish yellow-sepia particle composure. PDGF, TGF-βand TIMP-1 are mainly positioned in the endochylema. Completely observe each slice and casually select five representative high power field (10×40) to respectively measure the percentage and integral optical density of the positive section and the integral optical density of the positive section, taking the average integral optical density value of the positive section as the measurement value of this slice.2) The serum adopts ELISA method to operate in strict accordance with the instruction on the kit and read the absorbance value (OD value) for each hollow with the ELIASA in 450mm place, draw the standard curve to get the concentration of PDGF,TGF-βand TIMP-1 in three groups of serum.4 Statistical analysisConducted the statistics analysis with SPSS 17.0 software package and each group of data is indicated with x±s. The single factor variance analysis is adopted for the comparison among each measurement data for many groups and LSD-t is used to inspect for the comparison between two factors in the group. The Pearson relevant analysis is adopted for the relevance analysis among each indicator. The inspection standard is a=0.05.Results1 PDGF's representation in the endometrium and the serumPDGF has been represented in three groups of endometrium tissues, mainly concentrated on the endochylema of the endometrium glandular epithelium. PDGF's representation in endometrium tissues in the observation group is higher that that in the comparison group, among that, the PDGF's representation in endometrium tissues in severe adhesion group is higher that that in mild to moderate adhesion group. The difference has the statistic significance (P<0.05).PDGF has been tested in three groups of serum. PDGF's representation in the serum in the observation group is higher that that in the comparison group, among that, the PDGF's representation in serum in severe adhesion group is higher that that in mild to moderate adhesion group. The difference has the statistic significance (P<0.05).2 TGF-β's representation in the endometrium and the serumTGF-βhas been represented in three groups of endometrium tissues, mainly concentrated on the endochylema of the endometrium glandular epithelium. TGF-β's representation in endometrium tissues in the observation group is higher that that in the comparison group, among that, the TGF-β's representation in endometrium tissues in severe adhesion group is higher than that in mild to moderate adhesion group. The difference has the statistic significance (P<0.05).TGF-βhas been tested in three groups of serum. TGF-β's representation in the serum in the observation group is higher than that in the comparison group, among that the TGF-β's representation in serum in severe adhesion group is higher than that in mild to moderate adhesion group. The difference has the statistic significance (P<0.05).3 TIMP-1's representation in the endometrium and the serumTIMP-1 has been represented in three groups of endometrium tissues, mainly concentrated on the endochylema of the endometrium glandular epithelium. TIMP-1's representation in endometrium tissues in the observation group is higher than that in the comparison group, among that, the TIMP-1's representation in endometrium tissues in severe adhesion group is higher than that in mild to moderate adhesion group. The difference has the statistic significance (P<0.05).TIMP-1 has been tested in three groups of serum. TIMP-1's representation in the serum in the observation group is higher than that in the comparison group, among that the TIMP-1's representation in serum in severe adhesion group is higher than that in mild to moderate adhesion group. The difference has the statistic significance (P<0.05). 4 The relevance analysis of PDGF, TGF-βand TIMP-1 in IUA patients' endometrium tissuesMake the relevance analysis for the average integral optical density value represented by PDGF and TIMP-1 in endometrium tissues in the observation group, the result indicates:it is positive relevance between the two factors (r=0.661, P< 0.05). Make the relevance analysis for the average integral optical density value represented by TGF-βand TIMP-1 in endometrium tissues in the observation group, the result indicates:it is positive relevance between the two factors (r=0.661, P< 0.05). Make the relevance analysis for the average integral optical density value represented by TGF-βand TIMP-1 in endometrium tissues in the observation group, the result indicates:it is positive relevance between the two factors (r=0.630, P< 0.05).Conclusions1 PDGF, TGF-βand TIMP-1 may have significant role in IUA's pathogenesis;2 PDGF, TGF-βand TIMP-1 are related to the severity of IUA;3 PDGF, TGF-βand TIMP-1 may jointly participate in IUA's formation through mutual effects.