Expression Of SFlt-1 Signaling Pathway And The Early Onset Preeclampsia Correlation

ObjectivePreeclampsia( PE) is a serious threat to the health of mother and child, and its a group of disease charactors include hypertension, proteinuria, edema, multiple tube dysfunction. Additionally, the incidence rate is about 5%, and mortality is about 10%, and PE is a major cause of perinatal maternal and fetal infant mortality. The medical profession will be preeclampsia before 34 weeks ago called early-onset preeclampsia(EPE), onset later than 34 weeks are called late-onset preeclampsia(LPE). Early-onset preeclampsia has the characteristics of early disease,severe illness, more influence on mother and fetus, more complications, maternal and neonatal outcome worse prognosis.So it is particularly important finding out the causes, mechanisms, pathological processes, development law of the disease, effective prevention and treatment. Early-onset preeclampsia has become a hot topic in the medical field. At present, the treatment is given priority to with symptomatic treatment of early-onset preeclampsia. Effective molecular target for the treatment has become a clinical problems remaining to be solved.Placental growth factor(PLGF) is a vascular endothelial growth factor(VEGF) family member. PLGF combined with the vascular endothelial growth factor receptor 1(Flt-1) and play angiogenesis, promoting trophoblast cell growth, proliferation, invasion role, and this biological process can be blocked by soluble vascular endothelial growth factor receptor 1(s Flt-1). s Flt-1 and Flt-1 competitive binde VEGF( including PLGF).s Flt-1 completely blocked the role of VEGF and also blocked the angiogenic activity of PLGF, causing angiogenesis disorders, vascular wall integrity and permeability. s Flt-1,PLGF have a strong antagonistic effect. It has been reported that the expression of s Flt-1 is increased in hypertension, diabetes, multiple tumor. Some scholars have proposed to predict disease with s Flt-1 as a marker for preeclampsia. PI3 K / AKT pathway is involved in cell proliferation, differentiation, apoptosis and regulating glucose transport and other biological functions. Studies have shown that this pathway participate in activities as the development of human vascular, metabolic diseases and cancer, treatment and prognosis. These studies strongly suggest that s Flt-1 is associated with the PI3 K / AKT pathway and it could become targets for molecular therapy of early-onset preeclampsia. Based on this, We have completed the study of the expression of s Flt-1,PLGF in early-onset preeclampsia serum, placenta, umbilical cord and have completed the study of the expression of s Flt-1 with the PI3K/AKT pathway correlation by human umbilical vein endothelial cell. Methods⑴Collected 39 cases of early onset severe preeclampsia in Xijing Hospital from September 2012 to August 2014(ESPE group); Collected 45 cases of early onset mild preeclampsia(EMPE group); Collected 42 cases of normal pregnancy in the same period,and the same gestational age(NP group).We collected the patients blood, umbilical cord blood and the placenta during childbirth. We determined the content of s Flt-1 and PLGF in serum and cord blood by ELISA and time-resolved fluorescence immunoassay;The expression of s Flt-1 m RNA and PLGF m RNA in placenta were detected by real-time quantitative PCR; The expression of s Flt-1 and PLGF protein in placenta were detected by Western-blot.⑵Human umbilical cord specimens collected from normal pregnancy. Cultured human umbilical vein endothelial cells by trypsin digestion method. Used to detect CK18 to identify cells by immunofluorescence.⑶Established HUVECS hypoxia model.Detected HIF-1a by Western-blot method and TUNEL staining apoptosis. Hypoxic cells MTT assay performed to determine the time of the subsequent hypoxic hypoxia experiments.⑷Real-time quantitative PCR were used and Western-blot were used to detect s Flt-1, PI3Kp85α and p-AKT(ser473) in normal human umbilical vein endothelial cells, hypoxia 24 hours and one hour LY294002 role before hypoxia 24 hours.⑸By cell flow cytometry’s umbilical vein endothelial cells cultured in different ways under hypoxic cycle and apoptosis. Results⑴Early onset severe preeclampsia serum content of s Flt-1 the average number is 534.5 ± 22.6ng L, PLGF content is 34.9±8.7ng/L; early-onset mild preeclampsia serum content of s Flt-1 is 398.0±47.7 ng / L, the content of PLGF the average number is 51.6±10.4ng / L; the same period, with normal pregnancy gestational maternal serum levels of s Flt-1 average number is 212.0±31.9 ng / L, PLGF levels is 83.3±9.3ng / L. SPSS17.0 software using single factor analysis of variance, P <0.05, three groups were statistically significant. Early onset severe preeclampsia umbilical cord blood in serum s Flt-1 the average number is 331.2±40.4ng / L, PLGF levels mean is 29.0± 6.3ng / L; early onset of mild preeclampsia umbilical cord serum s Flt-1 mean content is 192.0±27.7 ng / L, PLGF levels mean is 41.3±5.0ng / L; the same period, with normal pregnancy gestational cord blood serum s Flt-1 content in the average number is 98.6 ± 13.9 ng / L, PLGF levels mean is 68.8±8.1ng / L. Three groups were statistically significant. Three groups placenta s Flt-1, PLGF in m RNA and protein levels are different, P <0.05.⑵Successfully cultured in vitro passaged human umbilical vein endothelial cells were cultured to identify cell-specific expression of CK18, proven reliable cell model building can be used for subsequent experiments.⑶The m RNA and protein levels s Flt-1, PI3Kp85α and p-AKT(ser473) were cultured in normal HUVECS low expression; in hypoxia 24 hours HUVECS expression; LY294002 effect 1 hour before 24 hours of hypoxia in HUVECS found that blocking the PI3 K / AKT pathway, while the expression of s Flt-1 has also undergone a reduction, P <0.05, the difference was statistically significant. Also confirmed by flow cytometry umbilical vein endothelial cells hypoxia occurred G0 and G1 phase arrest, reducing the percentage of S phase, increased apoptosis, while import LY294002 may weaken hypoxia on cells of this inhibition. conclusions Flt-1 in the serum of patients with early-onset preeclampsia, compared with normal umbilical cord and placenta in pregnant control expression, and as the disease worsens and increased expression; PLGF in the serum of patients with early-onset preeclampsia, umbilical cord blood and placenta relatively low expression in normal pregnancy control, and as the disease worsens and reduced expression. Explain its involvement in early-onset preeclampsia development, can be used as indicators of serum and histology of the disease.s Flt-1, PI3Kp85α and p-AKT(ser473) in HUVECs hypoxia model have both been highly expressed, indicating tissue hypoxia may be associated with early onset preeclampsia there is some intrinsic link, may be involved in early-onset preeclampsia the pathogenesis. PI3 K / AKT pathway inhibitor LY294002 role HUVECs1 hours before 24 hours under conditions of hypoxia expression of the three factors were reduced, indicating that PI3K/AKT pathway may be involved in the pathogenesis of early onset pre-eclampsia may be an effective therapeutic target molecule.