The Correlation Between The Concentration Of IL-6,IL-8 In Maternal Serum And The Expression Of MMPs In Fetal Membranes In Pregnancy Complicated With PROM

Objective:PROM (premature rupture of membranes) was a common complication in obstetric research, which had heavily puzzled the health of mother and fetus. In recent years, the incidence ratio of PROM was more and more high. So, many researchers were devoted to studying the etiology of PROM in order to deduce the incidence and mortality rate of fetus. The objection of the study was to explore the influence of enzyme and cytokine to PROM. The experiment method of immunohistochemistry(IHC) was to detect the expression and distribution of MMP-9 and TIMP-1 in fetal membranes. The enzyme linked immunosorbent assay (ELISA) was to detect the concentration of IL-6,IL-8 in maternal serum. It was wanted to know whether there were relationships between MMP-9 and IL-6,IL-8. All of these would provide a new idea for etiology of PROM and the theory basement for clinical therapy.Methods:There were 58 patients selected from obstetrical word of Second Affiliated Hospital Hebei Medical University. We divided them into three groups as follows:Ⅰgroup:21 gravidas, complicated with PROM, including Ⅰ1 group (n=8, with chorioamnionitis),Ⅰ2 group (n=13, without Chorioamnionitis). Ⅱgroup:17 gravidas, complicated with pPROM , including Ⅱ1 group (n=7, with chorioamnionitis) and Ⅱ2 group (n=10, without chorioamnionitis). Ⅲ group: 20 gravidas, normal controls. All patients had no physical and surgical diseases and other pregnancy complication. Anconeal venous blood would be abstracted from all samples during no uterine contrast before labor, separating blood serum, protecting it in -70℃ refrigerator. The membranes were collected from the point of rupture of membranes about 2cm×2cm×0.5cm, Using saline to clusis bloodstain, fixed by 10% neutral formalin and embeded in paraffin wax. Method:Detecting the concentration of IL-6,IL-8 with ELISA, procedures were as follows:1 the standard sample was diluted to 1000,500,250,125,62.5,31.25,15.625,0pg/ml. 2 took out of the blade of antibody from enclose pocket. 3 except blank pore, put the samples and different concentration of standard samples into corresponding pores, hatching 30min in 37℃. 4 washed the bland for 4 times. 5 except blank pore, added employment fluid after biotinyliring antibody, hatching 60min in 37℃. 6 washed blank for 4 times. 7 except blank pore, added enzyme linked employment fluid, hatching 30min in 37℃. 8 washed blank for 4 times. 9 added coloration, hatching 12min in 37℃ and blocking the light ray. 10 added terminal liquid, detecting every pore's absorbtion in wave length of 450nm. After stainned by HE, observing the structure of membranes. The experiment method of immunohistochemistry (IHC) was to detect the expression and distribution of MMP-9 and TIMP-1 in each group. Concrete procedures were as follows:1 After depparaffinization the paraffin section was performed in 3% hydrogen peroxide solution (H2O2), hatching for 10min to inactivate endogenous peroxidase. 2 antigen plerosis adopted with microwave, MMP-9 and TIMP-1 plerosised differently for 20min, 15min. 3 normal goat serum blocked antigen, hatching 10min in room temperature. 4 Added antibody-Ⅰ, overnight in 4℃. 5 Added biotin making antibody-Ⅱ, hatching 30min in 37℃. 6 Added horseradish enzyme labeling chain enzyme aridin, hatching 30min in 37℃. 7 MMP-9 showed colors for 3min and TIMP-1 for 2min by DAB, tap water terminated sections, counter stained with hematoxylin for 5min, Hydrochloric acid alcohol differentiation 20 seconds. Alcohol desiceation, hyalined by dimethylbenzene. After sealed sheet by gum, observed it.Results:1 Histological observation:In all PROM, those complicated with infection was 40.54%, without infection was 59.46%. In tPROM, those complicated with infection was 38.09%, middle and heavy Chorioamnionitis was 4.7%, slight Chorioamnionitis was 33.3%. In pPROM, those complicated with infection was 41.18%, middle and heavy Chorioamnionitis was 11.8%, slight Chorioamnionitis was 29.4%. In PROM, complicated with slight Chorioamnionitis, the inflam...