胎膜早破与SERPINH1基因多态性的相关性

Correlation between premature rupture of membrane and gene polymorphism of SERPINH1

目的探讨SERPINH1基因启动子区-656位点的单核苷酸多态性与胎膜早破的相关性。方法选择2010年1月—2012年10月在上海市第八人民医院明确诊断为早产胎膜早破(PPROM)患者56例(PPROM组,孕期<37周)、足月胎膜早破(TPROM)患者74例(TPROM组)、自然分娩的早产产妇67名(自然早产组,孕期<37周,未发生胎膜早破)和足月自然分娩的产妇134名(对照组)。抽取各组产妇外周血提取DNA,以聚合酶链反应扩增带有目的单核苷酸多态性的基因片段,并对产物进行测序。采用Taqman-MGB技术检测各组中SERPINH1基因-656位点单核苷酸多态性分布-656T和-656C基因型频率,采用相对荧光素酶活性测定检测热休克蛋白47(HSP47)mRNA在各组胎膜组织中的表达,采用Western印迹法检测HSP47蛋白在各组胎膜组织中表达,采用苏木精-伊红染色观察各组胎膜病理学变化情况。结果自然早产组、PPROM组和TPROM组中-656T基因型频率均显著高于对照组(P值均<0.05),而-656C基因型频率均显著低于对照组(P值均<0.05);HSP47mRNA、HSP47蛋白的表达水平均显著低于对照组(P值分别<0.05、0.01)。与对照组比较,自然早产组、PTPROM组和PPROM组的胎膜结构均发生了明显的改变,具体表现为胎膜连接层变宽,羊膜层纤维蛋白减少,部分滋养细胞发生凋亡,以及核碎裂等。结论 SERPINH1基因-656位点的单核苷酸多态性与TPROM的发生有关。

Objective To explore the correlation between premature rupture of membrane （PROM） and single nucleotide polymorphism （SNP） sites of SERPINH1. Methods A total of 56 cases of preterm premature rupture of membrane （PPROM, duration of pregnancy less than 37 weeks）, 74 term premature rupture of membrane （TPROM）, 67 premature birth （duration of pregnancy less than 37 weeks） and 134 normal full-term delivery, who were admitted to our hospital between January 2010 and October 2012, were enrolled in the present study. Peripheral blood of the parturients was collected. Gene fragments of SNPs were amplified by polymerase chain reaction （PCR） and all products were sequenced. The distribution of SNP genotypes on -656 genetic locus of SERPINH1 genes and the gene frequency of -656T and -656C in each group were detected with Taqman-MGB technology. The mRNA and protein levels of heat shock protein 47 （HSP47） were determined by real timepolymerase chain reaction （RT-PCR） and Western blotting analysis, respectively. The pathological changes of membranes in different groups were demonstrated by Hematoxylin-eosin （H-E） staining. Results The gene frequency on -656T genetic locus in premature birth, PPROM and TPROM groups were significantly higher than that in normal full-term delivery group （all P 〈 0. 05）, while the gene frequency on -6560 genetic locus were significantly lower than that in normal full-term delivery group （all P〈O. 05）. Compared with those in the normal full-term delivery group, the mRNA and protein levels of HSP47 in the other three groups were significantly decreased （P〈0.05, 0.01）. H-E staining results showed that fetal membranes assumed obviously pathologic changes in preterm labor, TPROM and PPROM groups; the connection layer of membranes broadened, the fibrin ofamniotic layer decreased, apoptosis and nuclear fragmentation appeared in some trophocytes. Conclusion SNP genotypes on -656 genetic locus of SERPINH1 gene have a strong correlation with PROM.