【作者】 李敏；

【导师】 石珍；

【作者基本信息】 南昌大学 ， 妇产科， 2007， 硕士

【摘要】 目的:对子癎前期患者的母胎转运的游离DNA进行定量测定,探讨转运的母胎游离DNA与子癎前期的关系及其在子癎前期疾病的发展和预测中的价值。方法:采用聚合酶链式反应(polymerase chain reaction,PCR)方法,利用谷胱甘肽S转移酶M1基因(glutathione S-transferase M1,GSTM1)、人血管紧张素转换酶基因(angiotensin-converting enzyme genes,ACE)基因多态性对同期住院分娩男婴的子癎前期及正常妊娠的母婴病例对进行筛选。通过实时荧光定量PCR(fluorescence quantitative PCR,FQ-PCR)方法利用特异性Y性别决定区(sex-determining region Y,SRY)基因及GSTM1、ACE基因多态性分别对入选病例对母血浆中胎儿游离DNA及脐血浆中母体游离DNA进行定量测定。同时利用放射免疫法对入选病例母血中绒毛膜促性腺激素(β-chorionic-gonadotropinhormone,β-HCG)浓度进行测定及相关比较。结果:1、共有52对母婴病例对入选进行FQ-PCR检测,其中实验组中子癎前期轻度组12对,重度组15对,正常对照组25对。2、52对病例对母血浆中均检测出胎儿游离DNA,实验组胎儿游离DNA浓度均值与对照组相比增高,子癎前期重度较轻度组增高,组间差异有显著性(P＜0.001)。3、脐血浆中检测出母体游离DNA,实验组共10例,对照组8例,实验组浓度高于对照组,子癎前期重度较轻度组增高。各组间浓度差异有统计学意义(P＜0.05)。4、实验组中子痼前期血β-HCG浓度值与对照组相比较,组间有显著性差异(P＜0.001),子癎前期母血β-HCG浓度高于正常妊娠。5、妊娠母血浆中游离胎儿DNA浓度、脐血浆中母体游离DNA浓度与母血β-HCG水平呈正相关(P＜0.01),转运的胎儿游离DNA浓度与母体游离DNA浓度无明显相关性(P＞0.05)。结论:1、妊娠高血压子癎前期转运的母胎游离DNA浓度高于正常妊娠,可能与胎盘滋养层和合体滋养层细胞凋亡增加有关。DNA增多意味着子癎前期与胎盘损伤有关,导致母儿交换增多。2、妊娠中转运的母、胎游离DNA浓度与子癎前期具有相关性,可能可以作为一项预测和监测子癎前期的临床指标。3、母胎转运游离DNA浓度与母血β-HCG水平相关,可用于子癎前期的临床预测和监测,并说明DNA代谢紊乱与子癎前期有相关性。更多还原

【Abstract】 Objective To quantitative analyze the bidirectional transfer within fetomaternal plasma free DNA and study the relationship with preeclampsia.To analyze the effects of the bidirectional free DNA traffic in the development and prognosis of preeclampsia.Methods Using a polymerase chain reaction(PCR)assay to screen the informative mother-baby pairs of which the pregnant women who carrying male babies and delivered in hospital,including 60 examples of preeclampsia(study group)and normal pregnancy(control group)respectively.The transfer from mother-to-fetus or fetus-to-mother was determined by detecting the inserting/deletion polymorphisms involving the glutathione S-transferase M1(GSTM1)and angiotensin-converting enzyme genes(ACE).The fetus-to-mother transfer of plasma DNA in pregnant women was studied using a fluorescence quantitative PCR(FQ-PCR)assay for sex-determining region Y(SRY)gene.For mother-to-fetus transfer,FQ-PCR assays for the insertion/deletion polymorphisms involving the GSTM1 and ACE genes were used.At the same time,radioimmunity assay was used to detect theβ-chorionicgonadotropin hormone(β-HCG).Results①A total of 52 informative mother-baby pairs were selected in all of 120 pairs.In the study group,there were 12 pairs of the mild degree of preeclampsia and 15 pairs of the severe degree of preeclampsia.The control group included 25 informative mother-baby pairs.②In the plasma fraction,fetal DNA was detected in 100%of maternal plasma(52 of 52).Compared with the control group,the mean fractional concentration in the group of the mild and severe degree of preeclampsia appeared significantly difference among groups(P＜0.001).The free fetal DNA concentration in preeclampsia was higher than which in the normal pregnancy.③Of the 27 informative mother-baby pairs,maternal DNA was detected only 10 pairs,including 4 pairs of mild degree of preeclampsia and 6 pairs of severe degree of preeclampsia.But 8 pairs in the control group were detected the maternal DNA.In the study group,the median fractional concentration was higher than the control group.The difference among groups was significance(P＜0.05).The free maternal DNA concentration in preeclampsia was higher than which in the normal pregnancy.④In comparision with normal pregnancy,the concentration ofβ-HCG in preeclampsia was significantly higher(P＜0.001).⑤Both the mother-to-fetus or fetus-to-mother transfer of plasma DNA had positive relationship withβ-HCG(P＜0.01).But there appeared to be no relationship between the fetus-to-mother and mother-to-fetus transfer of plasma DNA(P＞0.05).Conclusions①The concentration of the bidirectional transfer of feto-maternal plasma DNA in preeclampsia was higher than normal pregnancy.It might have the correlation with the increasing apoptosis of trophoblastic cells in placenta.So the increasing of free DNA might mean that the damage of placenta had relationship with preeclampsia,and resulted the increasing changes between fetus and mother at last.②The bidirectional transfer of feto-maternal free DNA have the relationship with preeclampsia.These results also raise the possibility that measurement of circulating DNA may prove useful as a marker for the diagnosis and/or monitoring of preeclampsia.③The concentration of the bidirectional transfer of feto-maternal plasma DNA had relationship with the concentration ofβ-HCG.Both of them could be used as the index of prediction and/or monitoring of preeclampsia.It suggested that preeclampsia was associated with metabolism disturbances of circulating DNA.更多还原