| ObjectivePPROM(preterm premature rupture of membrane)closely related to intrauterine infection,and may causes serious influence inthe outcomes of maternal and fetal.TRAF6 plays an important role in the regulation of inflammatory pathway in the pathogenesis of intrauterine infection.Recently,it has been suggested that MST4 can inhibit the activity of TRAF6 and play a role in inhibiting inflammation.In this study,we detected the expression of TRAF6 and MST4 in placenta tissue of patients with preterm premature rupture of membranes,and explored the mechanism and clinical significance of the interaction between them.And discussed the pathogenesis and clinical significance of the interaction between the two and its premature rupture of membranes and intrauterine infection.And meantime by analyzing the relationship between the expression of TARF6 and MST4 in the placenta of premature rupture of membranes with the outcome of pregnancy,to explore the value of TRAF6 and MST4 to predictparental outcome.Methods1.90 pregnant women who delivered at Subei People’s Hospital obstetrics and gynecology hospital of Jiangsu provincefrom May 2015 to May2016 are included in this study.Among them,60 cases complicated with PPROM,the rest 30 cases returned to normal control group.The data of pregnancy outcomes were collected,including gestational age,maternal birth,body temperature,body temperature before delivery,peripheral white blood cell count,neutrophil count,calcitonin and postpartum antibiotic use,Puerperium infection rate,and perinatal complications.2.All of the membranes were stained with HE test,then staining for intrauterine infection group and non infection groupaccording to the results of HE.The expression of TRAF6 and MST4 was detected by IHC(immunohistochemistry)and WB(western blot)Western blot protein imprinting in all placental tissues.Image-pro-plus 6 software was used to detect the number of positive cells and optical density.60 cases of premature rupture of membranes were analyzed according to the expression intensity of TRAF6 and MST4 in placental tissue.SPSS23.0 software was used for statistical analysis.Results1.Pathological results of membrane tissue:in the normal control group,there were 3 cases of intrauterine infection,accounting for the total number of cases of 10%(3/30),and 27 cases were not associated with intrauterine infection.PPROM group in 43 cases with intrauterine infection,accounting for the total number of cases of 71.66%(43/60),17 cases without intrauterine infection.2.General information:Using One-way ANOVA method analyze the general information between the three groups,the normal control group of 30 cases of preterm premature rupture of membranes in 17 cases with intrauterine infection group,PPROM group 43 cases with intrauterine infection.There was no statistically difference in subjects of age,gravidity,parity,Apgar score,BMI,maternal temperature,maternal peripheral blood white cell count,neutrophil count,procalcitonin value among three groups.The gestational age of the PPROM group was less than that of the normal control group(F=46.41,P<0.05),and the weight of newborn infants corresponding lighter in PPROM group(F=31.73,P<0.05).However,there was no significant difference in the gestational age and neonatal weight in preterm premature rupture of membranes combined with intrauterine infection and intrauterine infection.).3.Result of the relationship between time interval of membranes ruptured to delivery and intrauterine infection:60 cases in PPROM group were divided into three groups according to the time interval of membranes raptured to delivery,less than 12 hours in the intrauterine infection group in 11 cases,10 cases of 12-24 hours,more than 24 hours in 22 cases.There were 7 cases with no intrauterine infection in less than 12 hours,12-24 hours in 5 cases,and more than 24 hours in all cases.There was no significant difference between the two groups in the time interval after membranes ruptured(P=0.29).4.IHC results(1)The expression of TRAF6 was mainly localized in the nucleus of trophoblast,including 43 cases of PPROM with intrauterine infection group,the positive expression of TRAF6 in 42 cases(97.67%),of which 21 cases were strongly positive(48.84%),12 cases were moderate positive expression(27.91%),9 cases weakly positive(20.93%);in 17 cases of PPROM with intrauterine infection group,TRAF6 positive expression in 7 cases(41.18%),including 1 cases of moderate positive expression(5.88%),6 cases were weak positive(35.29%);30 cases of normal control group,the positive expression of TRAF6 12 cases(40%),including 2 cases of moderate positive expression(6.67%),10 cases weakly positive(33.33%).Using χ2 method to detect IHC results of three groups,the expression of PPROM with intrauterine infection of TRAF6 was significantly higher in the placental group and normal control group,PPROM patients without intrauterine infection group(χ2 =47.84,p<0.05;χ2 =32.87,P<0.05).And the group of PPROM patients without intrauterine infection and normal control group showed no significant abnormality(χ2=0.29,P=0.58).(2)the expression of MST4 in the normal placenta was observed in the nuclei and cytoplasm.The MOD values of normal control group,PPROM combined with intrauterine infection group and PPROM combined with intrauterine infection group were 0.03336±0.011,0.03393±0.012,0.05630±0.012.Data were analyzed by SPSS,the three group homogeneity of variance(p=0.99),using one-way ANOVA analysis showed that the expression of MST4 in placenta of the PPROM with intrauterine infection group was significantly higher than normal control group and the PPROM without intrauterine infection group(F=42.69,P<0.05).(3)Correlation analysis of TRAF6 and MST4 expression in different placental tissues by Spearman rank correlation analysis.The results showed that there was a positive correlation between the expression of TRAF6 and the expression of MST4 in placenta tissue(rs=0.69,p<0.05),in other words,the stronger the expression of TRAF6,the stronger the expression of MST4.5.Western blot protein imprinting resultsExpression of TRAF6 and MST4 in normal control group,PPROM with intrauterine infection group and PPROM without intrauterine infection group were detected by Western blot blotting.The relative gray level of TRAF6 in the normal control group and PPROM without intrauterine infection group and the PPROM with intrauterine infection group were 2.02±0.25,1.94 ± 0.18,3.34 ± 0.34.The relative gray level of TRAF6 was significantly higher than that of the normal control group and the PPROM without intrauterine infection group(F=256.01,P<0.05),while there was no significant difference between the normal control group and the PPROM without intrauterine infection group.The relative gray level of MST4 in the normal control group and PPROM without intrauterine infection group and the PPROM with intrauterine infection group were 0.29±0.04,0.27±0.03,1.55±0.25.The expression of MST4 in the PPROM with intrauterine infection group was significantly higher than that of the normal control group and the PPROM without intrauterine infection group(F=568.53,P<0.05),while there was no significant difference between the normal control group and the PPROM without intrauterine infection group.6.The relationship between the expression of TRAF6/MST4 and pregnancy outcomesThe expression of TRAF6 and MST4 were not correlated with the time of membranes ruptured,maternal postpartum fever,neonatal.sepsis and perinatal mortality(P>0.05),while them were associated with high neonatal NICU rate,and increase the incidence of neonatal pulmonary infection and pathologic jaundice(P<0.05).Conclusions1.TRAF6 and MST4 are participated in the PPROM with intrauterine infection and it development,but the specific mechanism is not yet clear.2.The increase of expression of TRAF6 and MST4 indicate high possibility to get neonatal asphyxia,pulmonary infection or pathological jaundice. |
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