HLA-G单核苷酸多态性与重度子痫前期的关联研究

The Association Study of HLA-G Single Nucleotide Polymorphisms and Severe Pre-eclampsia

目的:检测人类白细胞抗原(human leukocyte antigen,HLA)-G单核苷酸多态性(SNPs)位点在重度子痫前期患者及正常孕妇中的分布,了解重度子痫前期患者HLA-G SNPs的变化,以期进一步明确HLA-G在子痫前期免疫发病机制中的作用。方法:收集重度子痫前期组67例和对照组37例血液样本,提取基因组DNA,选取HLA-G外显子(Exon)2、3为实验研究区域,合成荧光标记引物,聚合酶链反应(PCR)、测序进行片段序列SNPs分析。统计分析所测SNPs位点与重度子痫前期的关联程度。结果:在HLA-G Exon2发现2个SNPs,为C374T及G476A;Exon3发现3个SNPs,为C810T、C859A及G954A。HLA-G Exon2、3 SNPs在重度子痫前期组及正常孕妇组间的分布差异无统计学意义(均P>0.05)。结论:(1)在HLA-G Exon2、3共发现5个SNPs位点,均为同义突变,与重度子痫前期无相关性。(2)C374T和G954A为新发现的HLA-G SNPs,向db SNP提交后获得NCBI Assay Id(ss#)119994701和119994702,Reference SNP Id(rs#):rs77474325和rs75783850。

Objective： To detect the distribution of HLA-G SNPs which is one of the susceptibility genes of pre-eclampsia in severe pre-eclampsia patients and normal pregnant women, to understand the change of HLA-G SNPs in the patients with severe pre-eclampsia. Methods： The blood samples were collected from 67 cases of severe pre-eclampsia group and 37 cases of control group, the genomic DNA of samples were extracted, the Exon-2 and Exon-3 of HLA-G SNPs were chosen to evaluate through sequencing, the fluorescent primers were synthesized and PCR was proceeded, the fragments obtained from PCR were sequenced and analyzed to find SNPs. The correlation of SNPs and severe pre-eclampsia was analyzed by statistical analysis. Results： Two SNPs, C374 T and G476 A were found in the Exon 2 of HLA-G. Three SNPs, C810 T, C859 A and G954 A were found in Exon 3. There was no significant difference in the distribution of the SNPs between pre-eclampsia and normal pregnancies. Conclusions：（1） Five SNPs were found in the HLA-G Exon 2 and 3, which were not related with severe pre-eclampsia, these SNPs were synonymous mutations.（2）C374T and G954 A were newly discovered HLA-G SNPs which were submitted to the db SNP and obtained NCBI Assay Id（ss#） 119994701 and 119994702, and the Reference SNP Id（rs#）：rs77474325 and rs75783850.