The Study Of ERAP2 Gene Polymorphism And Plasmic Value Of IL-32 In Preeclampsia

Pregnancy-induced hypertension is the most common complication of pregnancy, the incidence rate is 3-4% all over the world, 5-10% abroad, 5-8% in the United States and 9.4% domestic. Preeclampsia is associated with substantial maternal and fetal morbidity and mortality. Pre-eclampsia is a pregnancy-specific disease, clinically manifested as the mother syndrome (hypertension and proteinuria, with or without a multi-system dysfunction) or fetal syndrome (fetal growth restriction, oligohydramnios and oxygenation abnormalities). There continues to be active debate on the origins of this disease, although many scholars have put forth a lot of theories, but most have not withstood the test of time.The most accepted theories are as follows: Uterus-placental ischemia and hypoxia,coagulation dysfunction, vascular endothelial cell dysfunction, cardiovascular maladaptation, immune imbalance, genetic susceptibility, nutritional excess,and so on.Now widely recognized that pregnancy-induced hypertension is due to a number of different genetic factors influencing susceptibility gene or (and) the same susceptibility gene related to more than one sites, interacting with environmental factors, resulting disease. Vascular endothelial cell injury is an important part of disease.In 2009, Johnson MP and his colleagues used GeneSniffer program inquiring 5q QTL of pre-eclampsia patients of the Australian and the Norwegian, analyzing 10 candidate genes. They found a genetic association between ERAP2 SNP (rs2549782) of the Australian cohort with 140 preeclampsia patients (including 20 patients with eclampsia) and 90 control group (normal blood pressure without proteinuria) and ERAP2 SNP( rs17408150) with the Norwegian families having 1139 preeclampsia patients. They predicted the SNPs possibly and probably damage the 3-D ERAP2 protein structure and function, respectively.The missense rs2549782 SNP also resides within and juxtaposes the second Glutamic acid residue (E) in the highly conserved zinc-binding HEXXH(X)18E motif. This motif is a distinguishing characteristic of the zinc-dependent aminopeptidases and is essential in their enzymatic activity . To date, little information regarding the possible connection between ERAP2 gene polymorphisms and preeclampsia in a Chinese han population is available. We speculate ERAP2 may have a genetic linkage to this pregnancy-speciffic disorder.Most scholars believe that preeclampsia may be an excessive, uncontrolled inflammatory response, resulting in increasing of inflammatory cytokines. Information shows that women who are urethritis or periodontitis are more susceptible to preeclampsia. Premature birth, preeclampsia, miscarriage, fetal adverse outcomes are confirmed to be related with the inflammation. Inflammation is thought to be a an important factor in the pathological, CRP and IL-6 have acted as a predictor factor of preeclampsia . However, there continue unclear which inflammatory cytokine triggeres the response .IL-32 is a newly inflammatory cytokines, the relationship between preeclampsia and IL-32 arise our attention. This factor may have connection with atherosclerosis, coagulation, as well as cancer and endothelial inflammation. Lipid metabolism disorders, atherosclerosis and endothelial cell dysfunction have been confirmed in preeclampsia .Is IL-32 play a part in the occurrence and development of preeclampsia?IL-32 is found by Soo-Hyun Kim, who studied the high expression of IL-18 induced genes and discovered a cytokine-like gene. This gene had been named as NK4 (natural killer cell transcript 4) in 1992. Although the IL-32 have no homology with the other cytokine families,it can induce lymphocytes and mononuclear cells to produce many cytokines such as TNF-α, IL-1βand IL-8 and so on. Soo-Hyun Kim et al proved that IL-32 is an inflammatory cytokines. IL-32 plays an important role in many inflammatory diseases and cancer, such as rheumatoid arthritis, inflammatory bowel disease, Crohn's disease, gastric cancer and lung cancer and so on.We used molecular biological experiments to explore the relationship bewteen ERAP2 genetic polymorphism and pathogenesis of preeclampsia with normal pregnancy and preeclampsia; at the same time, we use immunological detection of a normal pregnancy and preeclampsia serum concentrations of IL-32, to investigate the role in the pathogenesis of preeclampsia , providing a theoretical basis for clinical early prevention, early diagnosis and early treatment .The present study is divided into two parts as follows: PartⅠThe association between ERAP2 gene polymorphism and severe preeclampsia【Abstract】Objective The purpose of the present study was to investigate the gene polymorphism between ERAP2(rs2549782, rs17408150) and Chinese Han population . Methods A case-control study was conducted involving 35 women with preeclampsia( study group )and 41 normotensive pregnant women(control group)in the third trimester.We uesd QIAmp DNA Blood mini kits to purife DNA from blood between preeclamptic and control groups , PCR amplifing the purposed bands,checking and purifing by Agarose gel electrophoresis ,sequencing by Beijing Genomics Institute(BGI) in shenzhen ,finally finding SNPs by Lasergene software and analysing by SPSS13.0. Results①the allele frequency of ERAP2 rs 17408150 between preeclampsia and normal controls is T.②When analysing rs17408150,we found a third SNP 36bp away,namely rs1423568,the ERAP2 rs1423568 allele frequencies for C% and T% were 37.14% and 62.86% in patients with preeclampsia, and 36.59% and 63.41% in normal pregnancies, respectively.χ2=0.003 , P=0.960 , OR= 1.024 , 95% CI 0.402-2.609.③the ERAP2 rs2548672 allele frequencies for T% and G% were 37.14% and 62.86% in patients with preeclampsia, and34.15% and65.85% in normal pregnancies, respectively.χ2= 0.074 ,P= 0.786 ,OR=1.140,95% CI 0.444-2.922. Conclusion Our work does not provide evidence in favor of ERAP2 SNP (rs2549782, rs1423568, rs17408150) being associated with preeclampsia and normal pregnant women in a Chinese han population.PartⅡThe study of IL-32 in serum of severe preeclampsia and controls【Abstract】Objective The purpose of the present study was to investigate the changes in plasma markers of IL-32 in preeclampsia and to evaluate their clinical significance and interactions in thepathogenesis of preeclampsia.Methods A case-control study was conducted involving 39 women with preeclampsia(study group) and 40 normotensive pregnant women(control group)in the third trimester.The plasma concentrations of interleukin-32(IL-6) were determined. Results The plasma comcentrations of IL-32 was significantly higher in the study group(789.07±167.70 ug/L) than those of the control(520.42±143.65 ug/L) (P<0.01,or P<0.05) Conclusion①IL-32 may play a role in the pathogenesis of PE.②The detection of serum levels of IL-32 in preeclampsia may provide a new way for early detection and treatment.③the serum levels of IL-32 is associated with the particulars of the disease. When the serum level of IL-32 increas, TNF-αis induecd, causing further endothelial cell injury of preeclampsia.