| Objective: To characterize the expression pattern of miR-3074-5p/p27 pathway molecules in placenta of pre-eclampsia,and to preliminarily explore its related mechanism with a view to identify some predictive molecules and to provide scientific and material bases for early intervention of pre-eclampsia.Methods: From September 2017 to March 2018,16 pregnant women with pre-eclampsia(PE)and 9 normal pregnant women(CTR)were enrolled in hospital delivery in the Second Hospital of Tianjin Medical University.Q-PCR were used to examine expression levels of miR-3074-5p,miR-146a-5p and miR-486-3p in placental samples.The expression levels of p27,p53,p57,Bcl-2,Bcl-G,Bax,cyclinD1,cyclinD2,CDK5,CDK4 and CDK2 in placental samples was determined by Western Blot.And expressions of cyclinD1,CDK2 and p27 proteins in placental tissues were localized by immunohistochemistry.Results: There was no significant difference in the age and gestational weeks of delivery between the PE group and CTR group.The BMI,systolic blood pressure and diastolic blood pressure in the PE group were significantly higher than that in the CTR group(31.63±4.24 vs 26.00±2.00;151.75±12.18 vs 112.22±14.15;103.94±9.31 vs 78.44±9.72;respectively,all P<0.01).There was no significant difference in hemoglobin,platelet count,D-dimer level,Cr,TBIL and ALT levels between the two groups before termination of pregnancy,whereas the levels of UA and BUN in the PE group were significantly higher than that in the CTR group(368.14±86.89 vs 274.60±43.69,P<0.01;3.93±1.12 vs 2.90±0.78,P<0.05;respectively).There was no significant difference in the newborn birthweight and the Apgar score(1,5,10 min),as well as the morbidity of neonatal asphyxia,premature,FGR/SGA and macrosomia/LGA,between the two groups.There was no significant difference in the morbidity of neonatal asphyxia,premature,FGR/SGA and macrosomia/LGA.The placenta of PE showed grayish red or dark red,soft,and the umbilical cord attached to the center or eccentrically.The length and diameter of placental umbilical cord were 30 ~ 55 cm and 0.5 ~ 1.2 cm respectively,some of the umbilical cord were slightly distorted and had staggered blood stasis.By HE staining,most of the PE placenta showed chorionic villi agglutination with chorionic space hyperemia or chorionic interstitial vasodilation,meanwhile placental nodules or syncytial nodules increased.Cellulose deposition with or without multiple focal infarction,focal calcification or multiple calcification was found in most cases.The q-PCR results showed that,the placental expression level of miR-3074-5p in PE group was significantly lower than that in CTR group(1.160e-006±2.521e-007 vs 4.286e-006±1.826e-006,P<0.05),whereas there was no significant difference in miR-146a-5p and miR-486-3p expression levels between the two groups.Western blot results showed that,The expression of p27 protein in PE group was significantly increased than that in CTR group(0.230±0.027 vs 0.117±0.022,P<0.01),but the expression levels of cyclinD1 and CDK4 proteins were significantly decreased(0.045±0.006 vs 0.085±0.018,P<0.05;0.258±0.053 vs 0.615±0.077,P<0.01).There was no significant difference between the two groups in the expression of p53,p57,cyclinD2,CDK5,CDK2,Bcl-2,Bcl-G and Bax.The immunohistochemical results showed that,the signals of cyclinD1,CDK2 and p27 were abundantly located in the cytoplasm of villus syncytiotrophoblast cells,and there were weak signals expressed in the cytotrophoblast cell.The number of villi in PE placenta was decreased,and the number of dyed particles,as well as the degree of staining of cyclinD1 were significantly reduced;whereas the number of dyed particles and degree of staining of p27 were significantly increased;however,there was no obvious change in the number of dyed particles and degree of staining of CDK2.Conclusion: In miR-3074-5p/p27/CCND1/CDK4 molecular pathway,the down-regulation expression of miR-3074-5p may be involved in the pathogenesis of PE by promoting the excessive apoptosis of placental syncytiotrophoblast. |
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