| Objective:The differently expressed proteins of preeclampsia and normal pregnancy in maternal serum were screened.Then,we detected the differently expressed proteins in other ways and explored their possible mechanism in preeclampsia,in order to find out the potential serum biomarkers for early prediction of preeclampsia.Methods:Patients with severe preeclampsia were collected from June 2015 to June 2016.All of them were diagnosed in the Affiliated Hospital of Qingdao University.30 cases of patients with severe preeclampsia were chosed as the case group.At the same time,30 cases of normal pregnant women were collected as the control group.The total proteins of the case group and the control group were extracted from blood serum,and then they were denatured,reduced,and enzymatic.Then they were identified by mass spectrometry after labed by iTRAQ.Quantitative proteomic analysis was carried out to analyze the differently expressed proteins,and the function of the differentially expressed proteins was analyzed by bioinformatics methods.At the same time,other 60 cases of patients with preeclampsia were collected,of which 30 cases were severe preeclampsia(SPE group),30 cases were mild preeclampsia(MPE group).30 cases of normal pregnant women were collected as the control group.Blood serums were separated and placenta tissues were collected.The differential protein HNP3 and PAPPA2 were tested in the three groups by enzyme linked immunosorbent assay and real-time fluorescence quantitative PCR.Result:(1)The differentially expressed proteins in serum were 234,of which 25 proteins were identified in SPE group more 1.6 times or 0.625 times less than the control group,and the T-test has proved that the differences had statistical significance.16 protein spots up regulated compared with the control group,and the other 9 proteins spots down regulated;(2)The HNP3 levels of the severe and mild preeclampsia group in serum were respectively(144.0±22.35)ng/m L ?(98.9±11.70)ng/m L,higher than that of the control group(83.8±11.71)ng/m L(P<0.0001?P=0.045,P<0.05),the differences between them were statistically significant.And the HNP3 levels of the severe group was higher than that of the mild group(P=0.0074,P<0.01),the difference also has statistical significance;(3)The relative expression of HNP3 m RNA of the severe and mild preeclampsia group in placental tissues were respectively(4.856±0.077)?(1.978±0.156),significantly higher than that of the control group(1.001±0.219)(P=0.001,P=0.032,P<0.05).The differences were statistically significant.Compared severe group with the mild group(P=0.015,P<0.05),there was also a statistically significant difference;(4)There was no correlations between the levels of HNP3 in serum and the relative expression of HNP3 m RNA in the placental tissues of patients with preeclampsia,the correlation coefficient was 0.554,P=0.626,P>0.05.There were no statistical significance;(5)The PAPPA2 levels of the severe and mild group of patients with preeclampsia in serum were respectively(549.1±36.68)pg/m L?(166.1±16.41)pg/m L,higher than that of the control group(138.1±9.05)pg/m L(P<0.0001?P=0.014,P<0.05).The differences between them were statistically significant.And the PAPPA2 levels of the severe group was higher than that of the mild group(P=0.0015,P<0.01),the difference also has statistical significance;(6)The relative expression of PAPPA2 m RNA of the severe group and mild group in the placental tissues of preeclampsia patients were respectively(13.136±0.230)?(4.523±0.282)(P<0.0001 ? P=0.015,P<0.05),higher than the control group(1.000±0.169),and the differences were statistically significant.Compared severe group with the mild group(P=0.001,P<0.01),there was also a statistically significant difference;(7)The level of PAPPA2 in serum of preeclampsia patients was positively correlated with the relative expression level of PAPPA2 m RNA in placenta,and the correlation coefficient was 0.944 and P=0.005,P<0.05.The correlation was statistically significant;(8)There was positive correlation between the levels of HNP3 and PAPPA2 in serum of preeclampsia patients,and the correlation coefficients was r=0.852,and P<0.0001.The correlation was statistically significant;Conclusions:(1)The application of iTRAQ technology combined with LC-MS/MS technology provides an effective technical support for the screening and identification of early signs of preeclampsia;(2)HNP3 and PAPPA2 may involved in the occurrence and development of preeclampsia;(3)HNP3 and PAPPA2 may have interaction in the pathogenic process of preeclampsia;(4)PAPPA2 may be the potential signs for early prediction of preeclampsia in serum. |
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