發(fā)布時(shí)間：2018-01-17 17:35

  本文關(guān)鍵詞：GCM1、Syncytin-1在子癇前期患者外周血及胎盤(pán)組織中的表達(dá)及其意義　出處：《河北醫(yī)科大學(xué)》2017年碩士論文　論文類(lèi)型：學(xué)位論文

  更多相關(guān)文章： 子癇前期 GCM1 Syncytin-1 滋養(yǎng)細(xì)胞 凋亡

【摘要】：目的:子癇前期(preeclampsia,PE)是妊娠期特有的疾病,可致圍產(chǎn)期母兒產(chǎn)生不良妊娠結(jié)局,是孕產(chǎn)婦和圍產(chǎn)兒病死率升高的主要原因,其基本病理生理變化為全身小血管痙攣。一直以來(lái),子癇前期的發(fā)病機(jī)制都是產(chǎn)科領(lǐng)域的研究熱點(diǎn),與胎盤(pán)淺著床、血管重鑄障礙、氧化應(yīng)激、凝血系統(tǒng)失衡、炎癥反應(yīng)及代謝異常等有關(guān),研究發(fā)現(xiàn)胎盤(pán)淺著床、血管重鑄障礙與胎盤(pán)滋養(yǎng)細(xì)胞的發(fā)育和功能異常有關(guān)。胎盤(pán)滋養(yǎng)細(xì)胞發(fā)揮正常功能需要多種蛋白因子共同作用,如:GCM家族、合胞素、胎盤(pán)生長(zhǎng)因子、可溶性血管內(nèi)皮生長(zhǎng)因子以及HtrA家族等。GCM1即GCMa,它是胎盤(pán)滋養(yǎng)細(xì)胞分化融合過(guò)程中所必需的轉(zhuǎn)錄因子,調(diào)控多種相關(guān)基因的表達(dá),參與胎盤(pán)滋養(yǎng)層細(xì)胞的增殖、分化以及融合。而Syncytin-l是由人內(nèi)源性逆轉(zhuǎn)錄缺陷病毒(HERV)系列中HERV-W編碼而形成的糖包膜蛋白,主要表達(dá)于胎盤(pán)合體滋養(yǎng)細(xì)胞的細(xì)胞膜和細(xì)胞質(zhì),GCM1作為轉(zhuǎn)錄因子調(diào)節(jié)Syncytin-1的表達(dá),二者相互作用共同參與滋養(yǎng)細(xì)胞增殖、分化、融合以及細(xì)胞凋亡。本研究通過(guò)檢測(cè)GCM1、Syncytin-1在正常妊娠孕婦與子癇前期患者外周血及胎盤(pán)組織中的表達(dá)水平,分析兩組間GCM1和Syncytin-1的表達(dá)差異以及二者之間的相關(guān)性,以探討其與子癇前期發(fā)病及嚴(yán)重程度的相關(guān)性,為子癇前期的早期診斷及病情評(píng)估提供依據(jù)。方法:1研究對(duì)象:依據(jù)《妊娠期高血壓疾病診治指南(2015)》子癇前期的診斷標(biāo)準(zhǔn),隨機(jī)選擇2015-06至2016-08河北醫(yī)科大學(xué)第二醫(yī)院收治的重度子癇前期患者130例,同期隨機(jī)選取正常孕婦61例,共納入病例191例,分為三組,早發(fā)組為早發(fā)型子癇前期(early onset severe preeclampsia)患者63例,晚發(fā)組為晚發(fā)型子癇前期(late onset preeclampsia)患者67例,正常對(duì)照組61例;早發(fā)組按孕周進(jìn)一步分層為30-31+6周共31例,32-34周共32例,晚發(fā)組按孕周進(jìn)一步分層為34+1-36+6周共32例,≥37周共35例。2標(biāo)本采集:在我院門(mén)診正規(guī)產(chǎn)檢并在我院剖宮產(chǎn)終止妊娠的初產(chǎn)婦在治療前取空腹靜脈血3ml,在室溫下2000rpm離心10分鐘,將分離出的血清置于ep管中,保存于-80℃冰箱,待測(cè)。胎盤(pán)娩出后30min內(nèi)收取胎盤(pán)母體面的中心部位,避開(kāi)鈣化灶,約1cm3大小組織,生理鹽水沖洗后用濾紙吸干水分,在10%福爾馬林溶液中室溫固定48h,并進(jìn)行常規(guī)的脫水、石蠟包埋等。3檢驗(yàn)指標(biāo):采用酶聯(lián)免疫吸附法(elisa)測(cè)定研究對(duì)象血清中g(shù)cm1、syncytin-1的濃度,采用免疫組織化學(xué)技術(shù)(immunohistochemistry,ihc)測(cè)定胎盤(pán)中g(shù)cm1、syncytin-1的表達(dá)水平,并記錄年齡、分娩孕周、孕次、身高、體重及血壓等指標(biāo)。采用he染色后觀察胎盤(pán)細(xì)胞滋養(yǎng)細(xì)胞、合體滋養(yǎng)細(xì)胞及絨毛間質(zhì)纖維化和絨毛血管病變情況;顯微鏡下觀察胎盤(pán)中g(shù)cm1、syncytin-1表達(dá)并人工計(jì)數(shù)進(jìn)行圖像分析。4數(shù)據(jù)分析:采用spss21.0統(tǒng)計(jì)軟件進(jìn)行數(shù)據(jù)分析。結(jié)果:1gcm1在對(duì)照組、早發(fā)組、晚發(fā)組血清中的表達(dá)水平分別平均為(11.61±2.27)ng/ml,(4.44±0.70)ng/ml、(7.27±0.97)ng/ml,子癇前期患者與正常孕婦相比,gcm1的表達(dá)水平明顯降低,且早發(fā)組降低更為明顯,差異有統(tǒng)計(jì)學(xué)意義(p0.05)。gcm1在30-31+6周組、32-34周組、34+1-36+6周組、≥37周組患者血清中的表達(dá)水平分別平均為(3.99±0.47)ng/ml,(4.87±0.50)ng/ml,(6.76±0.86)ng/ml,(7.99±0.71)ng/ml。經(jīng)統(tǒng)計(jì)學(xué)分析得出,30-31+6周組與32-34周組相比,gcm1表達(dá)水平降低,32-34周組與34+1-36+6組相比,gcm1表達(dá)水平呈下降趨勢(shì),34+1-36+6周組與≥37周組相比,表達(dá)水平也呈下降趨勢(shì),差異均有統(tǒng)計(jì)學(xué)意義(p0.05)。syncytin-1在對(duì)照組、早發(fā)組、晚發(fā)組血清中的表達(dá)水平分別平均為(10.47±0.62)ng/ml、(3.98±0.81)ng/ml、(7.73±1.14)ng/ml,子癇前期患者與正常孕婦相比,syncytin-1表達(dá)水平明顯降低,且早發(fā)組降低更為明顯,差異有統(tǒng)計(jì)學(xué)意義(p0.05)。syncytin-1在30-31+6周組、32-34周組、34+1-36+6周組、≥37周組患者血清中的表達(dá)水平分別平均為(3.65±0.40)ng/ml,(4.43±0.82)ng/ml,(6.81±0.87)ng/ml,(8.15±1.36)ng/ml。經(jīng)統(tǒng)計(jì)學(xué)分析得出,30-31+6周組與32-34周組相比,syncytin-1表達(dá)水平降低,32-34周組與34+1-36+6組相比,syncytin-1表達(dá)水平呈下降趨勢(shì),34+1-36+6周組與≥37周組相比,表達(dá)水平也呈下降趨勢(shì),差異均有統(tǒng)計(jì)學(xué)意義(p0.05)。2免疫組織化學(xué)顯示gcm1、syncytin-1在重度子癇前期患者和正常孕婦胎盤(pán)組織中均有表達(dá)。gcm1主要表達(dá)于胎盤(pán)合體滋養(yǎng)細(xì)胞的細(xì)胞漿,細(xì)胞滋養(yǎng)細(xì)胞的細(xì)胞核也有少量表達(dá);syncytin-1主要表達(dá)于胎盤(pán)合體滋養(yǎng)細(xì)胞的細(xì)胞膜和細(xì)胞漿。gcm1在對(duì)照組、早發(fā)組、晚發(fā)組的表達(dá)強(qiáng)度分別平均為(0.34±0.06)、(0.16±0.02)、(0.23±0.03);syncytin-1在對(duì)照組、早發(fā)組、晚發(fā)組的表達(dá)強(qiáng)度分別平均為(0.31±0.02)、(0.15±0.02)、(0.25±0.03),子癇前期患者與正常孕婦相比,gcm1、syncytin-1的表達(dá)強(qiáng)度均明顯降低,且早發(fā)組降低的更為明顯,差異均有統(tǒng)計(jì)學(xué)意義(均p0.05)。gcm1在30-31+6周組、32-34周組、34+1-36+6周組、≥37周組的表達(dá)強(qiáng)度分別平均為(0.14±0.01)、(0.17±0.01)、(0.22±0.02)、(0.24±0.03),經(jīng)統(tǒng)計(jì)學(xué)分析得出,30-31+6周組與32-34周組相比,gcm1表達(dá)強(qiáng)度明顯降低,32-34周組與34+1-36+6組相比,gcm1表達(dá)強(qiáng)度呈下降趨勢(shì),34+1-36+6周組與≥37周組相比,表達(dá)強(qiáng)度也呈降低趨勢(shì),差異均有統(tǒng)計(jì)學(xué)意義(p0.05)。syncytin-1在30-31+6周組、32-34周組、34+1-36+6周組、≥37周組的表達(dá)強(qiáng)度分別為(0.14±0.02)、(0.16±0.02)、(0.22±0.03)、(0.24±0.02),經(jīng)統(tǒng)計(jì)學(xué)分析得出,30-31+6周組與32-34周組相比,syncytin-1表達(dá)強(qiáng)度明顯降低,32-34周組與34+1-36+6組相比,syncytin-1表達(dá)強(qiáng)度呈下降趨勢(shì),34+1-36+6周組與≥37周組相比,表達(dá)強(qiáng)度也呈下降趨勢(shì),差異均有統(tǒng)計(jì)學(xué)意義(p0.05)。3he染色結(jié)果:對(duì)照組為正常妊娠孕婦的胎盤(pán)組織,可見(jiàn)絨毛成熟良好,絨毛及間質(zhì)血管豐富,形態(tài)正常,間質(zhì)少,絨毛表層的合體滋養(yǎng)層細(xì)胞細(xì)胞核排列較整齊;晚發(fā)型重度子癇前期組胎盤(pán)絨毛成熟不良,絨毛表層為細(xì)胞核排列紊亂的合體滋養(yǎng)層細(xì)胞,小動(dòng)脈平滑肌數(shù)量減少,可見(jiàn)玻璃樣變;早發(fā)型子癇前期組絨毛數(shù)量更少,絨毛明顯退變壞死,絨毛表面合體滋養(yǎng)層細(xì)胞大片融合。4外周血及胎盤(pán)組織中g(shù)cm1、syncytin-1的相關(guān)性分析顯示二者呈正相關(guān)(r=0.84,p0.001;r=0.84,p0.01)。結(jié)論:1重度子癇前期患者外周血中g(shù)cm1和syncytin-1表達(dá)水平降低,二者呈相關(guān)性,且早發(fā)組低于晚發(fā)組,發(fā)病孕周越小,表達(dá)水平越低。2重度子癇前期患者胎盤(pán)組織中GCM1和Syncytin-1均為低表達(dá),二者呈相關(guān)性,且早發(fā)組低于晚發(fā)組,發(fā)病孕周越小,表達(dá)水平越低。3重度子癇前期患者外周血和胎盤(pán)組織中GCM1和Syncytin-1表達(dá)呈正相關(guān),二者共同參與了子癇前期的病情發(fā)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.

【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R714.244

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