The Relationship Between Clusterin And Preeclampsia

【PURPOSE】1.To investigate the correlation between the expression of Clusterin(CLU)in serum and tissue and preeclampsia(PE);2.To analyze the relationship between serum CLU before 20 weeks of gestation and high risk factors,adverse outcomes,severity of preeclampsia,and routine laboratory indexes before 24 weeks of gestation,and to explore the predictive value of serum CLU for preeclampsia;3.To explore and analyze the role of CLU in the occurrence and development of preeclampsia,and provide new markers for the prediction and monitoring of preeclampsia.【METHODS】1.Participants: Pregnant women in Foshan Maternal and Child Health Hospital and Guangzhou Women and Children’s Medical Center were treated from September 2017 to June 2020.A total of 88 pregnant women with hypertensive disorder complicating pregnancy and 88 normal control women were enrolled in the nested case-control study.The serum samples were collected before 24 weeks of gestation.In addition,44 cases of serum in late pregnancy(22 cases in PE group and 22 cases in control group,matched by age and gestational age,1:1),10 cases of placental tissue in delivery(5 cases in PE group and 5 cases in control group,matched by age),and 4 cases of placental tissue in early pregnancy(including villus and decidua tissue).2.Research methods:1)Methods: enzyme linked immunosorbent assay(ELISA)was used to detect the expression level of CLU in serum of pregnant women;western blot and immunohistochemistry were used to detect the location and expression level of CLU in placental tissue of early and late pregnancy;fluorescence quantitative PCR was used to detect CLU m RNA and MMP-9 m RNA in placental tissue of late pregnancy.At the same time,transwell invasion assay was used to observe the effect of CLU protein on trophoblast invasion.2)Statistical methods: SPSS16.0 statistical software package was used to analyze and process the data.The measurement data were expressed by X ±s,and the comparison between the two groups was performed by t test or lost-sum test;the count data were expressed by frequency or rate,and the comparison was performed by chi square test.P < 0.05 was considered statistically significant.Logistic regression and receiver operating characteristic curve(ROC)were used to evaluate the predictive value of PE risk factors,routine blood indicators and CLU level.【RESULTS】1.demographic data and blood index changes in preeclampsia:1.1 High risk factors of PE: compared with the control group(n = 88),the proportion of pregnant women with scar uterus,and previous abortion / stillbirth history in PE group(n = 59)was higher(P < 0.05),and the relative risk RR(95% CI)of scar uterus and previous abortion / stillbirth history were 8.20(1.89-35.68)and 1.71(1.52-2.55).1.2 Adverse outcomes of PE: compared with the control group(n = 88),the incidence of adverse outcomes in PE group(n = 59)was higher,mainly manifested in the increased risk of hypertension in late pregnancy,oligohydramnios and cesarean section,as well as the occurrence ofpremature delivery,intrauterine distress,fetal growth retardation,low birth weight,small gestational age,neonatal respiratory distress syndrome and neonatal high mortality The incidence of bilirubinemia increased,with statistical difference(P <0.05).1.3 The blood indexes before 24 weeks of pregnancy in PE group: the average volume of red blood cells and hemoglobin content of red blood cells in PE group were lower than that of normal group(P < 0.05);the red blood cell count,distribution width CV of red blood cells,glutamyl-transferase,alanine aminotransferase,creatinine,urea and uric acid were higher than that of the normal group(P < 0.05).2.Correlation between CLU and preeclampsia2.1 The expression level of serum CLU: there was significant difference between PE group(435.80 ± 54.00 μg/ml)and normal control group(348.90 ±36.00μg/ml)before 24 weeks of gestation(P = 0.006),especially in severe preeclampsia(445.1±66.5μg/ml)(P =0.007).Before 24 weeks of gestation,the expression level of serum CLU increased between HP group(403.5 ± 57.3 μ g / ml),mild PE group(414.5 ± 102.1 μ g / ml)and severe PE group(445.1 ± 66.5 μ g / ml).In the serum of late pregnancy,the expression level of CLU in PE group(34.86±6.55μg/ml)was significantly higher than that in normal control group(19.18±4.13μg/ml)(P = 0.001).2.2 Expression of CLU in placenta: the expression level of CLU in villi was significantly higher than that in decidua in the first trimester of pregnancy(P = 0.001);in the third trimester of pregnancy,CLU was mainly located in the cytoplasm and nucleus of trophoblast cells,and the expression level of CLU in placenta of PE group was significantly higher than that in normal control group(P = 0.020).2.3 Correlation analysis: there was no significant correlation between serum CLU expression level before 24 weeks of gestation and high risk factors of PE,such as scar uterus,previous abortion / stillbirth history(P > 0.05).The expression level of CLU before24 weeks of gestation was correlated with adverse outcomes(pregnant women with hypertension in the third trimester and fetal distress).Higher concentration of CLU increased the incidence of both(P < 0.05).In PE pregnant women,there was no significant linear correlation between the expression level of serum CLU and routine blood indexes in the same period.2.4 The predictive value of CLU: the serum CLU before 24 weeks of gestation,statistically significant high-risk factors and routine blood indexes were included in the logistic multivariate regression equation,and the joint prediction model was obtained: y =-9.399 + 21.312 *(scar uterus)+ 22.559 *(abortion / stillbirth History)+ 0.143 *(creatinine)+ 0.003 *(CLU),and the prediction ROC curve of preeclampsia was made,and the area under the curve was 0.927(95% CI: 0.862-0.992),the best critical value of the model was 0.47,sensitivity: 0.800,specificity: 0.982.3.CLU is involved in the occurrence of preeclampsia3.1 Study on the mechanism of CLU involved in the occurrence of preeclampsia: in the placenta of preeclampsia,the expression of CLU m RNA increased(P = 0.010),while the expression of MMP-9 m RNA decreased(P = 0.007),and there was a negative correlation between them(r =-0.74,P < 0.05).3.2 In addition,50 ng / ml and 100 ng / ml of CLU protein solution could inhibit the invasion of HTR-8/SVneo cells.【CONCLUSION】1.PE is a specific disease of pregnant women,which is affected by many factors and can cause a variety of adverse maternal and infant outcomes.The routine blood index of PE pregnant women had some changes before onset.2.CLU can be used as a biomarker for preeclampsia.The high expression of serum CLU before 24 weeks of gestation was related to preeclampsia,its severity and adverse outcomes,but not to its high risk factors.And the combination of serum CLU before 24 weeks of gestation with high risk factors and routine blood indexes has a high early predictive value for preeclampsia.3.CLU may participate in the occurrence of preeclampsia by inhibiting the invasion of trophoblast.