GCM1、Syncytin-1在子痫前期患者外周血及胎盘组织中的表达及其意义

发布时间：2018-01-17 17:35

本文关键词：GCM1、Syncytin-1在子痫前期患者外周血及胎盘组织中的表达及其意义　出处：《河北医科大学》2017年硕士论文　论文类型：学位论文

更多相关文章： 子痫前期 GCM1 Syncytin-1 滋养细胞 凋亡

【摘要】：目的:子痫前期(preeclampsia,PE)是妊娠期特有的疾病,可致围产期母儿产生不良妊娠结局,是孕产妇和围产儿病死率升高的主要原因,其基本病理生理变化为全身小血管痉挛。一直以来,子痫前期的发病机制都是产科领域的研究热点,与胎盘浅着床、血管重铸障碍、氧化应激、凝血系统失衡、炎症反应及代谢异常等有关,研究发现胎盘浅着床、血管重铸障碍与胎盘滋养细胞的发育和功能异常有关。胎盘滋养细胞发挥正常功能需要多种蛋白因子共同作用,如:GCM家族、合胞素、胎盘生长因子、可溶性血管内皮生长因子以及HtrA家族等。GCM1即GCMa,它是胎盘滋养细胞分化融合过程中所必需的转录因子,调控多种相关基因的表达,参与胎盘滋养层细胞的增殖、分化以及融合。而Syncytin-l是由人内源性逆转录缺陷病毒(HERV)系列中HERV-W编码而形成的糖包膜蛋白,主要表达于胎盘合体滋养细胞的细胞膜和细胞质,GCM1作为转录因子调节Syncytin-1的表达,二者相互作用共同参与滋养细胞增殖、分化、融合以及细胞凋亡。本研究通过检测GCM1、Syncytin-1在正常妊娠孕妇与子痫前期患者外周血及胎盘组织中的表达水平,分析两组间GCM1和Syncytin-1的表达差异以及二者之间的相关性,以探讨其与子痫前期发病及严重程度的相关性,为子痫前期的早期诊断及病情评估提供依据。方法:1研究对象:依据《妊娠期高血压疾病诊治指南(2015)》子痫前期的诊断标准,随机选择2015-06至2016-08河北医科大学第二医院收治的重度子痫前期患者130例,同期随机选取正常孕妇61例,共纳入病例191例,分为三组,早发组为早发型子痫前期(early onset severe preeclampsia)患者63例,晚发组为晚发型子痫前期(late onset preeclampsia)患者67例,正常对照组61例;早发组按孕周进一步分层为30-31+6周共31例,32-34周共32例,晚发组按孕周进一步分层为34+1-36+6周共32例,≥37周共35例。2标本采集:在我院门诊正规产检并在我院剖宫产终止妊娠的初产妇在治疗前取空腹静脉血3ml,在室温下2000rpm离心10分钟,将分离出的血清置于ep管中,保存于-80℃冰箱,待测。胎盘娩出后30min内收取胎盘母体面的中心部位,避开钙化灶,约1cm3大小组织,生理盐水冲洗后用滤纸吸干水分,在10%福尔马林溶液中室温固定48h,并进行常规的脱水、石蜡包埋等。3检验指标:采用酶联免疫吸附法(elisa)测定研究对象血清中gcm1、syncytin-1的浓度,采用免疫组织化学技术(immunohistochemistry,ihc)测定胎盘中gcm1、syncytin-1的表达水平,并记录年龄、分娩孕周、孕次、身高、体重及血压等指标。采用he染色后观察胎盘细胞滋养细胞、合体滋养细胞及绒毛间质纤维化和绒毛血管病变情况;显微镜下观察胎盘中gcm1、syncytin-1表达并人工计数进行图像分析。4数据分析:采用spss21.0统计软件进行数据分析。结果:1gcm1在对照组、早发组、晚发组血清中的表达水平分别平均为(11.61±2.27)ng/ml,(4.44±0.70)ng/ml、(7.27±0.97)ng/ml,子痫前期患者与正常孕妇相比,gcm1的表达水平明显降低,且早发组降低更为明显,差异有统计学意义(p0.05)。gcm1在30-31+6周组、32-34周组、34+1-36+6周组、≥37周组患者血清中的表达水平分别平均为(3.99±0.47)ng/ml,(4.87±0.50)ng/ml,(6.76±0.86)ng/ml,(7.99±0.71)ng/ml。经统计学分析得出,30-31+6周组与32-34周组相比,gcm1表达水平降低,32-34周组与34+1-36+6组相比,gcm1表达水平呈下降趋势,34+1-36+6周组与≥37周组相比,表达水平也呈下降趋势,差异均有统计学意义(p0.05)。syncytin-1在对照组、早发组、晚发组血清中的表达水平分别平均为(10.47±0.62)ng/ml、(3.98±0.81)ng/ml、(7.73±1.14)ng/ml,子痫前期患者与正常孕妇相比,syncytin-1表达水平明显降低,且早发组降低更为明显,差异有统计学意义(p0.05)。syncytin-1在30-31+6周组、32-34周组、34+1-36+6周组、≥37周组患者血清中的表达水平分别平均为(3.65±0.40)ng/ml,(4.43±0.82)ng/ml,(6.81±0.87)ng/ml,(8.15±1.36)ng/ml。经统计学分析得出,30-31+6周组与32-34周组相比,syncytin-1表达水平降低,32-34周组与34+1-36+6组相比,syncytin-1表达水平呈下降趋势,34+1-36+6周组与≥37周组相比,表达水平也呈下降趋势,差异均有统计学意义(p0.05)。2免疫组织化学显示gcm1、syncytin-1在重度子痫前期患者和正常孕妇胎盘组织中均有表达。gcm1主要表达于胎盘合体滋养细胞的细胞浆,细胞滋养细胞的细胞核也有少量表达;syncytin-1主要表达于胎盘合体滋养细胞的细胞膜和细胞浆。gcm1在对照组、早发组、晚发组的表达强度分别平均为(0.34±0.06)、(0.16±0.02)、(0.23±0.03);syncytin-1在对照组、早发组、晚发组的表达强度分别平均为(0.31±0.02)、(0.15±0.02)、(0.25±0.03),子痫前期患者与正常孕妇相比,gcm1、syncytin-1的表达强度均明显降低,且早发组降低的更为明显,差异均有统计学意义(均p0.05)。gcm1在30-31+6周组、32-34周组、34+1-36+6周组、≥37周组的表达强度分别平均为(0.14±0.01)、(0.17±0.01)、(0.22±0.02)、(0.24±0.03),经统计学分析得出,30-31+6周组与32-34周组相比,gcm1表达强度明显降低,32-34周组与34+1-36+6组相比,gcm1表达强度呈下降趋势,34+1-36+6周组与≥37周组相比,表达强度也呈降低趋势,差异均有统计学意义(p0.05)。syncytin-1在30-31+6周组、32-34周组、34+1-36+6周组、≥37周组的表达强度分别为(0.14±0.02)、(0.16±0.02)、(0.22±0.03)、(0.24±0.02),经统计学分析得出,30-31+6周组与32-34周组相比,syncytin-1表达强度明显降低,32-34周组与34+1-36+6组相比,syncytin-1表达强度呈下降趋势,34+1-36+6周组与≥37周组相比,表达强度也呈下降趋势,差异均有统计学意义(p0.05)。3he染色结果:对照组为正常妊娠孕妇的胎盘组织,可见绒毛成熟良好,绒毛及间质血管丰富,形态正常,间质少,绒毛表层的合体滋养层细胞细胞核排列较整齐;晚发型重度子痫前期组胎盘绒毛成熟不良,绒毛表层为细胞核排列紊乱的合体滋养层细胞,小动脉平滑肌数量减少,可见玻璃样变;早发型子痫前期组绒毛数量更少,绒毛明显退变坏死,绒毛表面合体滋养层细胞大片融合。4外周血及胎盘组织中gcm1、syncytin-1的相关性分析显示二者呈正相关(r=0.84,p0.001;r=0.84,p0.01)。结论:1重度子痫前期患者外周血中gcm1和syncytin-1表达水平降低,二者呈相关性,且早发组低于晚发组,发病孕周越小,表达水平越低。2重度子痫前期患者胎盘组织中GCM1和Syncytin-1均为低表达,二者呈相关性,且早发组低于晚发组,发病孕周越小,表达水平越低。3重度子痫前期患者外周血和胎盘组织中GCM1和Syncytin-1表达呈正相关,二者共同参与了子痫前期的病情发展。
[Abstract]:Objective: preeclampsia (preeclampsia, PE) is a pregnancy specific disease, cause of perinatal adverse pregnancy outcomes, is a major cause of maternal and perinatal mortality, the basic pathophysiology of systemic small vessel spasm. Since the pathogenesis of preeclampsia are the hot research field of obstetrics with the shallow placental implantation, vascular remolding, oxidative stress, coagulation system imbalance, inflammatory reaction and abnormal metabolism related study found that shallow placental implantation, development and work of vascular remolding and placental trophoblast cells can abnormalities. Trophoblast cells function normally requires the joint action, a variety of factors such as: GCM protein family, syncytin, placental growth factor, soluble vascular endothelial growth factor and HtrA family of.GCM1 GCMa, which is the differentiation of trophoblast fusion transcription factor required in the process of, Regulate the expression of many genes involved in placental trophoblast cells, proliferation, differentiation and integration. The Syncytin-l is composed of human endogenous retroviruses (HERV) encoding sugar envelope protein HERV-W in the series and the formation of the cell membrane was mainly expressed in placental syncytiotrophoblast and cytoplasmic expression of GCM1, as a transcription factor regulating Syncytin-1, two the interaction involved in trophoblast cell proliferation, differentiation, apoptosis and cell fusion. This study through the detection of GCM1, the expression level of Syncytin-1 in patients with preeclampsia and normal pregnant women peripheral blood and placenta tissues, analyze the correlation between the expression differences between the two groups of GCM1 and Syncytin-1 and the two, to investigate the relationship with the onset and severity of preeclampsia, and provide the basis for early diagnosis and severity evaluation of preeclampsia. Methods: 1 subjects: according to the < pregnancy high Blood disease diagnosis and treatment guidelines (2015) "diagnostic criteria for preeclampsia, were randomly selected from 2015-06 to 2016-08 of the second hospital of Hebei Medical University, 130 cases of severe preeclampsia patients, randomly selected 61 cases of normal pregnant women, 191 cases were included and divided into three groups, the early group for early onset preeclampsia (early onset severe preeclampsia) patients 63 cases of late onset group for late-onset preeclampsia (late onset preeclampsia) in 67 cases, 61 cases of normal control group; the early-onset group according to gestational age 30-31+6 weeks were further divided into 31 cases of 32-34 weeks, a total of 32 cases of late onset group according to gestational age 34+1-36+6 weeks were further divided into 32 cases, more than 37 weeks a total of 35 cases of.2 were collected in our hospital: regular prenatal examination in our hospital cesarean section primipara before treatment, fasting blood 3ml, 2000rpm at room temperature and centrifuged for 10 minutes, the serum separated from the EP tube, stored in the -80 C The refrigerator, to be measured. For parts of maternal surface of placenta center within 30min after delivery of the placenta, avoid calcification, about the size of 1cm3 group, saline rinse after removing water with filter and fixed in 10% formaldehyde solution at room temperature and 48h, conventional dehydration, paraffin embedded and.3 test index: using enzyme linked immunosorbent assay method for determination of Gcm1 (ELISA) of serum, the concentration of syncytin-1, immunohistochemical technique (immunohistochemistry, IHC) determination of Gcm1 in placenta, the expression level of syncytin-1, and record the age, gestational age, pregnancy, height, weight and blood pressure. Observation of placental trophoblast cells by HE staining. The syncytiotrophoblast and villous interstitial fibrosis and villous vascular lesions; observation of Gcm1 in placenta under the microscope, the expression of syncytin-1 and manual counting image analysis.4 data analysis using spss21.0 statistical software Data were analyzed. Results: the 1gcm1 in the control group, the early group and late expression levels in serum were averaged for (11.61 + 2.27) ng/ml, (4.44 + 0.70) ng/ml, (7.27 + 0.97) ng/ml in patients with preeclampsia compared with normal pregnant women, the expression level of Gcm1 decreased significantly, and the early group decreased more significantly, the difference was statistically significant (P0.05).Gcm1 in 30-31+6 weeks group, 32-34 weeks group, 34+1-36+6 weeks group, 37 weeks group than the expression level in the serum of the patients respectively (3.99 + 0.47) ng/ml, (4.87 + 0.50) ng/ml, (6.76 + 0.86) ng/ml, (7.99 + 0.71) ng/ml. by statistical analysis, 30-31+6 week group compared with the 32-34 group, the expression level of Gcm1 decreased, compared with 32-34 group and 34+1-36+6 group, the expression level of Gcm1 decreased, compared with 34+1-36+6 group and more than 37 weeks, the expression level also decreased, the differences were statistically significant (P0.05) in the control of.Syncytin-1 early onset group. Group, late expression levels in serum were averaged for (10.47 + 0.62) ng/ml, (3.98 + 0.81) ng/ml, (7.73 + 1.14) ng/ml in patients with preeclampsia compared with normal pregnant women, the expression level of syncytin-1 decreased significantly, and the early-onset group decreased more significantly, the difference was statistically significant (P0.05).Syncytin-1 in 30-31+6 weeks group, 32-34 weeks group, 34+1-36+6 weeks group, 37 weeks group than the expression level in the serum of the patients respectively (3.65 + 0.40) ng/ml, (4.43 + 0.82) ng/ml, (6.81 + 0.87) ng/ml, (8.15 + 1.36) ng/ml. obtained by statistical analysis, 30-31+6 week group compared with 32-34 weeks group, the expression level of syncytin-1 decreased, compared with 32-34 group and 34+1-36+6 group, the expression level of syncytin-1 decreased, compared with 34+1-36+6 group and more than 37 weeks, the expression level also decreased, the differences were statistically significant (P0.05).2 immunohistochemistry showed that Gcm1 and syncytin-1 in severe preeclampsia The cytoplasm expressed.Gcm1 mainly expressed in placenta of patients and normal placenta, trophoblast cells also have a small amount of expression; syncytin-1 is mainly expressed in placental syncytiotrophoblast cell membrane and the cytoplasm of the.Gcm1 cells in the control group, the early group, the expression intensity of the late-onset group respectively (0.34 + 0.06), (0.16 + 0.02), (0.23 + 0.03); syncytin-1 in the control group, the early group, the expression intensity of the late-onset group respectively (0.31 + 0.02), (0.15 + 0.02), (0.25 + 0.03), patients with preeclampsia compared with normal pregnant women, Gcm1, expression the syncytin-1 were significantly decreased, and the premature group decreased more significantly, there were statistically significant differences (P0.05).Gcm1 in 30-31+6 weeks group, 32-34 weeks group, 34+1-36+6 weeks group, over expression intensity of 37 week group respectively (0.14 + 0.01), (0.17 + 0.01), (0.22 + 0.02) (0.24 + 0.03), By statistical analysis, 30-31+6 week group compared with 32-34 group, the expression intensity of Gcm1 decreased significantly, compared with 32-34 group and 34+1-36+6 group, the expression intensity of Gcm1 decreased, compared with 34+1-36+6 group and more than 37 weeks, the expression intensity also decreased, the differences were statistically significant (P0.05.Syncytin-1) in 30-31+6 group, 32-34 weeks group, 34+1-36+6 weeks group, over expression intensity of 37 weeks group respectively (0.14 + 0.02), (0.16 + 0.02), (0.22 + 0.03), (0.24 + 0.02), by statistical analysis, 30-31+6 week group compared with 32-34 group, the expression intensity of syncytin-1 decreased significantly, compared to 32-34 week group and 34+1-36+6 group, the expression intensity of syncytin-1 decreased, compared with 34+1-36+6 group and more than 37 weeks, the expression intensity also decreased, the differences were statistically significant (P0.05).3he staining: control group normal pregnancy placenta chorionic villi, mature and good Well, the villi and interstitial vascular rich, normal morphology, interstitial, syncytiotrophoblast cell surface villi were arranged in order; late onset severe preeclampsia placental villi villi surface is mature, disordered arrangement of the nuclei of syncytiotrophoblast cells, arteriolar smooth muscle to reduce the number of visible hyaline; early onset preeclampsia the number of villi less obvious degeneration and necrosis of villi, villous syncytiotrophoblast large surface Gcm1 fusion.4 in peripheral blood and placenta tissue, analysis the correlation between syncytin-1 showed that two had positive correlation (r= 0.84, p0.001; r=0.84, P0.01). Conclusion: 1 patients with severe preeclampsia in the peripheral blood of Gcm1 and syncytin-1 expression decreased the two are related, and early-onset group were lower than the late group, the incidence of gestational age is small, the lower the level of expression of.2 in severe preeclampsia group GCM1 and Syncytin-1 were low expression, The two were correlated, and the early onset group was lower than the late onset group. The smaller the incidence of gestational age, the lower the expression level. The expression of GCM1 and Syncytin-1 in peripheral blood and placenta tissues of.3 patients was positively correlated, and the two participated in the development of preeclampsia.

【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R714.244

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