Analysis On Gene Polymorphism Of Vascular Endothelial Growth Factor Susceptibility And Severe Preeclapsia

Objectives1 To investigate the relationship between VEGF 936C/T gene polymorphism and severe preeclampsia.2 To investigate the relationship between serum VEGF levels and severe preeclampsia.3 To investigate the relationship between VEGF 936C/T gene polymorphism and serum VEGF levels.Methods1 PatientEighty-four patients with preeclampsia served as study group, which patients are in line with "Obstetrics and Gynecology" (7th edition) and severe diagnostic criteria for preeclampsia.And seventy-one normal pregnant women were selected as control group. All of the subjects had no past diabetes mellitus, chronic hypertension and severe heart, liver and kidney dysfunction; had no other obstetric complications and medical complications; non-smoking, alcoholism, drug abuse or mental illness history.2 DNA and serum preparationAll of the subjects were taken in the early morning fasting blood before delivery 8ml, 3ml 2% EDTA 0.3ml anticoagulant,low-permeability separation of white blood cell. DNA was extracted by the chloroform/iso-amyl alcohol and saved in -80°c for genetic polymorphism preserved detection. 3ml plus separating gel/coagulant anticoagulant,serum was separated and prepared for detections of VEGF concentrations and biochemical markers.3 VEGF 936C/T gene polymerphism testThe DNA rewarming to room temperature, amplification polymerase chain reaction (PCR) of DNA, primer sequences using PCR-RFLP analysis. PCR amplified products obtained with the enzyme NlaⅢwater bath at 37℃overnight, digested products on 2% agarose gel (EB staining) were electrophoresis, VEGF 936C/T gene polymorphism genotyping results: the CC of VEGF 936C/T gene polymorphism was the wild-type and showed a 401bp of the amplified fragment because of no restriction sites. TT homozygous mutation has a restriction site showed two 264bp, 137bp fragment, and CT heterozygote for the mutation, indicating 401bp, 264bp, 137bp fragment 3; the use of software SHEsis is consistent with Hardy-Weinberg genetic equilibrium; The distribution of differences of the genotype and allele between severe preeclampsia group and the normal control group were compared by using chi-square test, and the allele was associated with severe preeclampsia group. P values less than 0.05 was considered statistically significant. 4 Detection of serum VEGF concentrationThe frozen serum was rewarmed to room temperature. The concentration of serum VEGF was detected by nusing enzyme-linked immunosorbent assay (ELISA). The test of homogeneity of variance t test was used for the differences of levels of serum VEGF between severe preeclampsia pre-group and the normal control group. The correlation of VEGF 936C/T gene polymorphism and serum VEGF levels was analysised by the test of homogeneity of variance using the test after. Calculation of bivariate correlation analysis using the VEGF 936C/T gene polymorphism and serum VEGF levels correlated.P values less than 0.05 was considered statistically significant.5 Patients with severe preeclampsia related factors VEGF concentration The serum of patients with severe preeclampsia, VEGF concentration and age, body mass index, gravidity, parity, gestational age, blood pressure and other indicators index Pearson correlation analysis to study patients with severe preeclampsia related factors VEGF concentration.6 Severe preeclampsia unconditional Logistic multivariate analysis To whether the occurrence of severe pre-eclampsia as the dependent variable, with unconditional Logistic regression analysis of multiple factors on the incidence of severe preeclampsia influence.Results1 Severe preeclampsia group systolic blood pressure, diastolic blood pressure, body mass index were higher than the normal control group.The difference was statistically significant(P <0.01).2 Compared with normal control group AST,ALP, ALB, TP, BUN, Cr, UA, LDH,CK,TG,LDL-C,Chol,HDL-Cand severe preeclampsia.the differences were statistically significant (P <0.01).3 Compared with age, body mass index, gestational age, biochemical parameters and blood pressure,VEGF 936C/T polymorphism in the genotype had no statistically difference (P> 0.05).4 The CC,CT+TT genotype frequency of VEGF 936 C/T gene polymorphism in severe preeclampsia group were 58.3%,46.7%; C and T allele frequency were 79.2%,20.8%. In the normal control group, the CC, CT+TT genotype frequency of VEGF 936 C/T gene polymorphism were78.9%,21.1%; C and T allele frequency were 89.4% and 10.6%.5 VEGF 936 CT+TT genotype of severe preeclampsia group was higher than that of the normal control group,and the difference was statistically Significance (P<0.05);VEGF 936CC genotype of severe preeclampsia group was lower than that of the normal control group,and the difference was statistically significance (P<0.05);VEGF 936 T allele of severe preeclampsia group was higher than that of the normal control group, and the difference was statistically significant (P<0.05); VEGF 936 C allele of severe preeclampsia group was lower than that of the normal control group, and the difference was statistically significant (P<0.05).6 The serum of patients with severe preeclampsia, VEGF concentration and age, body mass index, gravidity, parity, gestational age, blood pressure and other indicators index Pearson correlation analysis showed that patients with severe preeclampsia, SBP with age in serum VEGF concentration , DBP, AST, ALT, TGe, Chol, LDH correlated (P <0.05).7 The serum VEGF levels of severe preeclampsia and the normal control group were (344.95±192.79) pg / ml and (268.88±236.80) pg/ml, and the difference was statistically significant (P<0.05).8 VEGF 936C/T gene polymorphism and serum VEGF levels of correlation analysis showed the difference was not statistically significant (P>0.05).9 Logistic regression analysis revealed that serum VEGF level,VEGF genotype ,Cr, ,LDHwere important risk factors of severe preeclampsia.10 The CT+TT genotype of patients with severe preeclampsia, incidence of CC genotype of 2.732 times.Conclusions1 VEGF 936 C/T gene polymorphism and severe preeclampsia are relevant, the cases with CT/TT genotypes have high risk of preeclampia.2 Levels of VEGF of patients with severe preeclampsia significantly increased, The increase of maternal serum VEGF with severe preeclampsia and the severity of disease progression. 3 The relationship between VEGF 936C/T gene polymorphism and serum VEGF levels were found.