天堂国产午夜亚洲专区-少妇人妻综合久久蜜臀-国产成人户外露出视频在线-国产91传媒一区二区三区

子癇前期患者胎盤(pán)組織中miR-155及CXCR4的表達(dá)及意義

發(fā)布時(shí)間:2018-07-26 14:44
【摘要】:子癇前期(Preeclampsia,PE)是妊娠期常見(jiàn)內(nèi)科并發(fā)癥——妊娠期高血壓疾病中的一種類型,在產(chǎn)科較為常見(jiàn),其診斷要點(diǎn)是孕婦于妊娠20周后首次出現(xiàn)收縮壓≥140mm Hg和(或)舒張壓≥90mm Hg,加上以下新發(fā)條件之一:蛋白尿(≥300mg/24h,或隨機(jī)尿蛋白+),母體器官功能障礙(包括腎功能不全、肝臟損害、神經(jīng)或血液系統(tǒng)并發(fā)癥)及子宮胎盤(pán)功能不全(可能導(dǎo)致胎兒生長(zhǎng)受限)。從全球來(lái)看,在低收入和中等收入的國(guó)家,每年有超過(guò)100萬(wàn)婦女死于妊娠相關(guān)疾病,大約10%的女性在妊娠期間患有高血壓,其中并發(fā)子癇前期的占2%~8%?紤]到胎盤(pán)介于母體和胎兒之間,是維持胎兒宮內(nèi)生長(zhǎng)發(fā)育的重要器官,基于胎盤(pán)從母體娩出后PE臨床癥狀能夠較快緩解或可以自愈的事實(shí),推測(cè)子癇前期的發(fā)病與胎盤(pán)的病理生理變化密切相關(guān)。大多學(xué)者認(rèn)為胎盤(pán)的結(jié)構(gòu)較為復(fù)雜,滋養(yǎng)細(xì)胞是其主要的細(xì)胞類型,而滋養(yǎng)細(xì)胞功能異常會(huì)導(dǎo)致對(duì)孕婦血管的侵襲障礙,即造成子宮螺旋動(dòng)脈重塑不足,導(dǎo)致子宮胎盤(pán)血流灌注減少、胎盤(pán)缺血缺氧,進(jìn)而釋放一些細(xì)胞因子進(jìn)入母體循環(huán),損害孕婦血管內(nèi)皮細(xì)胞。相關(guān)的發(fā)病學(xué)說(shuō)還涉及遺傳、營(yíng)養(yǎng)、炎癥及胰島素抵抗等方面,但截止到目前,子癇前期的具體發(fā)病機(jī)制仍不清楚。MicroRNA(miRNA)是一類長(zhǎng)度為19~25個(gè)核苷酸非編碼蛋白質(zhì)的單鏈RNA,在基因轉(zhuǎn)錄后水平抑制信使RNA(m RNA)的表達(dá)或致其降解。自從研究者成功識(shí)別了3707個(gè)成熟的miRNAs之后,miRNA數(shù)據(jù)庫(kù)得以建立,生物信息學(xué)分析表明人類超過(guò)30%的基因可能受miRNAs的調(diào)控,最近,越來(lái)越多的證據(jù)表明胎盤(pán)組織中異常表達(dá)的miRNAs與子癇前期的發(fā)病有關(guān)。趨化因子受體4(CXCR4)是趨化因子基質(zhì)細(xì)胞衍生因子-1(stromal cell-derived factor-1,SDF-1,又稱CXCL12)的特異性受體,兩者有較強(qiáng)的親和力,通常形成SDF-1/CXCR4生物軸發(fā)揮生理學(xué)效應(yīng)。有研究證實(shí),SDF-1/CXCR4軸不僅影響免疫細(xì)胞的活化、遷移和募集,還參與血管生成、造血功能、胚胎發(fā)育以及腫瘤的侵襲和轉(zhuǎn)移等多種進(jìn)程。另外有學(xué)者在癌癥相關(guān)的研究中發(fā)現(xiàn)CXCR4可能是miR-155的一個(gè)靶基因,miR-155通過(guò)調(diào)控CXCR4基因的表達(dá),影響癌癥的血管生成及癌細(xì)胞的功能,進(jìn)而參與癌癥的發(fā)生。但在子癇前期發(fā)病過(guò)程中,miR-155與CXCR4之間的表達(dá)及如何參與子癇前期的發(fā)病機(jī)制尚未見(jiàn)相關(guān)的研究報(bào)道。目的本研究旨在利用實(shí)時(shí)熒光定量PCR(Real-time PCR)的實(shí)驗(yàn)方法,檢測(cè)重度子癇前期組和正常對(duì)照組孕婦胎盤(pán)組織中miR-155和CXCR4 m RNA的表達(dá)水平,采用鏈霉親和素—生物素—過(guò)氧化物酶復(fù)合物(即SABC法)免疫組織化學(xué)的實(shí)驗(yàn)方法、使用高通量檢測(cè)工具——組織芯片對(duì)子癇前期組和正常對(duì)照組孕婦胎盤(pán)組織中CXCR4蛋白的定位及表達(dá)進(jìn)行分析,初步探討胎盤(pán)組織中miR-155和CXCR4的表達(dá)以及與子癇前期發(fā)病的關(guān)系。資料與方法1一般資料1.1 Real-time PCR研究對(duì)象選取2015年11月至2016年02月在河南省婦幼保健院產(chǎn)科住院,臨床確診為重度子癇前期、最終以剖宮產(chǎn)術(shù)分娩的30例孕婦的胎盤(pán)組織作為實(shí)驗(yàn)組(s PE組),同期因社會(huì)因素行剖宮產(chǎn)術(shù)分娩的30例健康孕婦的胎盤(pán)組織作為正常對(duì)照組(N組)用于檢測(cè)m RNA的表達(dá)。1.2胎盤(pán)組織芯片研究對(duì)象同一課題組前期收集2007年12月12日至2010年12月31日在鄭州大學(xué)第三附屬醫(yī)院住院,臨床診斷為子癇前期、最終以剖宮產(chǎn)術(shù)分娩的80例孕婦的胎盤(pán)組織,同期因社會(huì)因素行剖宮產(chǎn)術(shù)分娩的58例健康孕婦的胎盤(pán)組織,由河南省婦幼健康轉(zhuǎn)化醫(yī)學(xué)工程實(shí)驗(yàn)室用于構(gòu)建一種高通量檢測(cè)工具——胎盤(pán)組織芯片,用來(lái)分析CXCR4蛋白的定位及表達(dá)。1.3研究對(duì)象診斷及納入標(biāo)準(zhǔn)子癇前期及其不同分類的診斷標(biāo)準(zhǔn)嚴(yán)格按照人民衛(wèi)生出版社出版的第八版《婦產(chǎn)科學(xué)》的診斷標(biāo)準(zhǔn),整個(gè)孕期血壓低于140/90 mm Hg,無(wú)尿蛋白出現(xiàn),無(wú)母體內(nèi)、外科及自身免疫性疾病等合并癥,無(wú)其他產(chǎn)科并發(fā)癥及胎兒異常情況等將納入正常對(duì)照組。本實(shí)驗(yàn)中所納入的全部研究對(duì)象均為單胎妊娠、剖宮產(chǎn)分娩。2方法采用實(shí)時(shí)熒光定量PCR技術(shù)從m RNA水平檢測(cè)實(shí)驗(yàn)組和正常對(duì)照組孕婦胎盤(pán)組織中miR-155及CXCR4的表達(dá)情況;采用Spearman方法分析實(shí)驗(yàn)組孕婦胎盤(pán)組織中miR-155和CXCR4 m RNA表達(dá)的相關(guān)性;采用鏈霉親和素—生物素—過(guò)氧化物酶復(fù)合物(即SABC法)免疫組織化學(xué)的實(shí)驗(yàn)方法從蛋白水平檢測(cè)河南省婦幼健康轉(zhuǎn)化醫(yī)學(xué)工程實(shí)驗(yàn)室前期成功構(gòu)建的胎盤(pán)絨毛滋養(yǎng)細(xì)胞(VCT)組織芯片(正常對(duì)照組42例,子癇前期56例)和絨毛外滋養(yǎng)細(xì)胞(EVCT)組織芯片(正常對(duì)照組29例,子癇前期47例)中CXCR4蛋白的表達(dá)及定位。3統(tǒng)計(jì)學(xué)方法實(shí)驗(yàn)過(guò)程中所得數(shù)據(jù)均采用SPSS 22.0統(tǒng)計(jì)軟件進(jìn)行分析。計(jì)量資料表示為均數(shù)±標(biāo)準(zhǔn)差((?)±s),使用2-△△Ct方法分析Real time-PCR實(shí)驗(yàn)結(jié)果。檢驗(yàn)數(shù)據(jù)正態(tài)分布及方差齊性后,兩者均符合者使用兩獨(dú)立樣本t檢驗(yàn)。相關(guān)分析采用Spearman相關(guān)分析法。免疫組化的實(shí)驗(yàn)結(jié)果為有序多分類的等級(jí)資料,使用秩和檢驗(yàn)進(jìn)行統(tǒng)計(jì),以α=0.05為檢驗(yàn)水準(zhǔn)。結(jié)果1實(shí)驗(yàn)組和正常對(duì)照組孕婦臨床特征的比較實(shí)驗(yàn)組(s PE組)孕婦年齡在22~35歲之間,平均為27.7±2.8歲,正常對(duì)照組(N組)孕婦年齡在20~32歲之間,平均為28.8±2.5歲。s PE組的分娩時(shí)體重指數(shù)(Body Msaa Index,BMI)、分娩孕周、收縮壓、舒張壓、平均動(dòng)脈壓、新生兒出生體重、尿蛋白分別為21.2±2.9 kg/m2、37.2±2.7周、169.5±17.6 mm Hg、112.4±15.7mm Hg、131.5±15.6 mm Hg、2210.6±905.1g、5.6±2.6 g/L。而正常對(duì)照組分別為22.2±2.4 kg/m2、38.7±0.8周、112.6±9.4 mm Hg、71.8±7.1mm Hg、85.6±7.5 mm Hg、3650.5±403.6g、0 g/L。在孕婦年齡、分娩時(shí)BMI及分娩孕周方面,兩組相比差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05);在收縮壓、舒張壓、平均動(dòng)脈壓及尿蛋白方面,實(shí)驗(yàn)組均顯著高于正常對(duì)照組,在新生兒出生體重方面,實(shí)驗(yàn)組低于正常對(duì)照組,且差異均有統(tǒng)計(jì)學(xué)意義(P0.05)。2 miR-155和CXCR4 m RNA在實(shí)驗(yàn)組和正常對(duì)照組孕婦胎盤(pán)組織中的表達(dá)miR-155、CXCR4 m RNA在實(shí)驗(yàn)組和正常對(duì)照組孕婦胎盤(pán)組織中的相對(duì)表達(dá)量依次分別為1.53±0.92、0.87±0.73;0.54±0.38、1.53±0.73,兩組比較差異具有統(tǒng)計(jì)學(xué)意義(P0.05)。3 miR-155和CXCR4 m RNA在實(shí)驗(yàn)組孕婦胎盤(pán)組織中表達(dá)的相關(guān)性miR-155在實(shí)驗(yàn)組孕婦胎盤(pán)組織中的相對(duì)表達(dá)量為1.53±0.92,CXCR4m RNA在實(shí)驗(yàn)組孕婦胎盤(pán)組織中的相對(duì)表達(dá)量為0.54±0.38,采用Spearman檢驗(yàn)對(duì)實(shí)驗(yàn)組孕婦胎盤(pán)組織中miR-155及CXCR4 m RNA進(jìn)行相關(guān)性分析,結(jié)果顯示:二者呈顯著負(fù)相關(guān)關(guān)系(r=㧟0.773,P0.05)。4 CXCR4蛋白在胎盤(pán)VCT和EVCT組織芯片中的表達(dá)及定位CXCR4蛋白在胎盤(pán)滋養(yǎng)細(xì)胞和絨毛外滋養(yǎng)細(xì)胞的細(xì)胞膜和細(xì)胞質(zhì)中均可見(jiàn)表達(dá)。通過(guò)對(duì)所有位點(diǎn)染色結(jié)果進(jìn)行分析顯示:胎盤(pán)VCT組織芯片中,CXCR4蛋白在子癇前期組的陽(yáng)性表達(dá)率為48.21%(27/56),正常對(duì)照組的陽(yáng)性表達(dá)率為83.33%(35/42),兩組相比差異有統(tǒng)計(jì)學(xué)意義(U=715.5,P0.05);胎盤(pán)EVCT組織芯片中:CXCR4蛋白在子癇前期組的陽(yáng)性表達(dá)率為48.94%(23/47),正常對(duì)照組為79.31%(23/29),兩組相比差異有統(tǒng)計(jì)學(xué)意義(U=362.0,P0.05)。結(jié)論重度子癇前期患者胎盤(pán)組織中miR-155可能通過(guò)調(diào)控CXCR4的表達(dá)參與子癇前期的發(fā)病。
[Abstract]:Preeclampsia (Preeclampsia, PE) is one of the common internal complications of pregnancy, which is one of the types of hypertensive disorders in pregnancy. It is more common in obstetrics. The main point of diagnosis is that after 20 weeks of pregnancy, the first occurrence of systolic pressure of more than 140mm Hg and (or) diastolic pressure is more than 90mm Hg, plus one of the following new conditions: proteinuria (> 300mg/24h, or random). Urinary protein +), maternal organ dysfunction (including renal dysfunction, liver damage, neurological or blood system complications) and uterine placenta dysfunction (which may lead to fetal growth restriction). Globally, more than 1 million women die of pregnancy related diseases in low and middle income countries, and about 10% of women are in pregnancy. Between the preeclampsia and the preeclampsia, 2%~8%. takes into account that the placenta is between the mother and the fetus, and is an important organ to maintain the fetal intrauterine growth and development, based on the fact that the PE clinical symptoms can be quickly relieved or self healing after the delivery of the mother body from the mother body. It is speculated that the pathogenesis of preeclampsia is closely related to the pathophysiological changes of the placenta. Most scholars believe that the placental structure is more complex, trophoblast is the main cell type, and the dysfunction of trophoblast will lead to the invasion of blood vessels in pregnant women, that is, the insufficiency of the uterine spiral artery, the decrease of the uterine placenta blood flow, the ischemia and hypoxia of the placenta, and the release of some cytokines into the mother body. Circulation, which damages the vascular endothelial cells of pregnant women. The related pathogenesis also involves heredity, nutrition, inflammation and insulin resistance. But at present, the specific pathogenesis of preeclampsia remains unclear..MicroRNA (miRNA) is a single strand RNA of a class of 19~25 nucleotide non coding proteins, which inhibits messenger R at the post transcriptional level. The expression or degradation of NA (m RNA). Since the researchers successfully identified 3707 mature miRNAs, the miRNA database has been established. Bioinformatics analysis shows that more than 30% of the human gene may be regulated by miRNAs. Recently, more and more evidence suggests that the abnormal expression of miRNAs in the placenta is associated with the onset of preeclampsia. Chemokine receptor 4 (CXCR4) is a specific receptor for chemokine matrix cell derived factor -1 (stromal cell-derived factor-1, SDF-1, and CXCL12). Both have strong affinity and usually form a SDF-1/CXCR4 biological axis to play physiological effects. Also involved in a variety of processes, such as angiogenesis, hematopoiesis, embryonic development, and tumor invasion and metastasis. Other scholars have found that CXCR4 may be a target gene for miR-155 in cancer related studies. MiR-155 affects the angiogenesis of cancer and the function of cancer cells by regulating the expression of CXCR4 gene, and then participates in the occurrence of cancer. However, the expression of miR-155 and CXCR4 and how to participate in the pathogenesis of preeclampsia have not been reported in the process of preeclampsia. The purpose of this study was to detect miR-155 and CXCR4 m in placental tissues of pregnant women with severe preeclampsia and normal control group by real time fluorescence quantitative PCR (Real-time PCR). The expression level of RNA, using the experimental method of streptomycin biotin peroxidase complex (SABC) immuno histochemistry, using high flux detection tool - the localization and expression of CXCR4 protein in placental tissue of preeclampsia and normal control group by high flux detection tool, and preliminarily discuss the MI in placenta tissue The expression of R-155 and CXCR4 and the relationship with preeclampsia. Data and methods 1 general data 1.1 Real-time PCR subjects were enrolled in obstetrics and Gynecology of Henan maternity and child health care hospital from November 2015 to 2016, and 30 cases of pregnant women with severe preeclampsia and cesarean section were selected as experimental group (s PE). The placental tissue of 30 healthy pregnant women in the same period as the normal control group (group N) was used to detect the expression of M RNA expression.1.2 placenta tissue chip. The same subject group was collected in the Third Affiliated Hospital of Zhengzhou University from December 12, 2007 to December 31, 2010, and the clinical diagnosis was pre eclampsia before the pre eclampsia. The placental tissue of 80 pregnant women with cesarean section and 58 healthy pregnant women with cesarean section due to social factors in the same period, and the placental tissue of 58 healthy pregnant women in the same period due to social factors, the placental tissue chip was used to analyze the location and expression of CXCR4 protein by the maternal and child health transformation Medical Engineering Laboratory of Henan province. The diagnostic criteria for the diagnosis and inclusion of standard preeclampsia and its different classifications were strictly according to the diagnostic standard of the eighth edition of Obstetrics and Gynecology published by the people's Health Press. The whole pregnancy blood pressure was lower than 140/90 mm Hg, no urinary protein appeared, no maternal body, surgical and autoimmune disease and other complications, and no other obstetric complications were found. The abnormal conditions of the fetus were included in the normal control group. All the subjects included in this experiment were single pregnancy. The.2 method of cesarean delivery was used to detect the appearance of miR-155 and CXCR4 in the placental tissues of the pregnant women and the normal control group by real time fluorescence quantitative PCR technique, and the Spearman method was used to analyze the experimental group. The correlation between the expression of miR-155 and CXCR4 m RNA in pregnant women's placenta and the immunohistochemical staining of streptomycin biotin peroxidase complex (SABC method) was used to detect the placental chorionic trophoblast (VCT) tissue microarray of the maternal and child health transformation Medical Engineering Laboratory of Henan province. The expression of CXCR4 protein in 42 cases of normal control, 56 cases of preeclampsia and EVCT (29 cases of normal control, 47 cases of preeclampsia) were analyzed by SPSS 22 statistical software. The measurement data were expressed as mean standard deviation ((?) + s), and 2- delta was used. The results of the Real time-PCR experiment were analyzed by the delta Ct method. After testing the normal distribution of the data and the homogeneity of variance, both of them used the two independent sample t test. The correlation analysis adopted the Spearman correlation analysis. The experimental results of the immunohistochemistry were the hierarchical data of hierarchical classification, the rank sum test was used for statistics, and the results were alpha =0.05 as the test level. 1 the experimental group and the normal control group of pregnant women's clinical characteristics of the experimental group (Group s PE) between the age of 22~35, the average age of 27.7 + 2.8 years old, the normal control group (group N) of pregnant women age between 20~32 years, the average of 28.8 + 2.5 years old.S PE group of birth weight index (Body Msaa Index, BMI), labor cycle, systolic pressure, diastolic pressure, mean arterial pressure The birth weight of the newborns was 21.2 + 2.9 kg/m2,37.2 + 2.7 weeks, 169.5 + 17.6 mm Hg, 112.4 + 15.7mm Hg, 131.5 + 15.6 mm Hg, 2210.6 + 905.1g, 5.6 + 2.6 g/L., while the normal control group was 22.2 + mm Hg. The difference between the two groups was not statistically significant (P0.05), and the experimental group was significantly higher than the normal control group in the aspects of systolic pressure, diastolic pressure, mean arterial pressure and urine protein. In the birth weight of the newborn, the experimental group was lower than the normal control group, and the difference was statistically significant (P0.05).2 miR-155 and CXCR4 m RNA in the real group (P0.05). The expression of miR-155, CXCR4 m RNA in the placental tissues of the pregnant women and the normal control group was 1.53 + 0.92,0.87 0.73 and 0.54 + 0.38,1.53 + 0.73 respectively in the experimental group and the normal control group. The two groups were statistically significant (P0.05).3 miR-155 and CXCR4 m RNA in the experimental group of pregnant women placenta group The relative expression of miR-155 expression in the placental tissue of the experimental group was 1.53 + 0.92, and the relative expression of CXCR4m RNA in the placental tissue of the experimental group was 0.54 + 0.38. The correlation analysis of miR-155 and CXCR4 m RNA in the placental tissue of the experimental group was analyzed by Spearman test. The results showed that the two were significantly negative. Expression of.4 CXCR4 protein (r=? 0.773, P0.05) in placental VCT and EVCT tissue microarray and localization of CXCR4 protein in the cell membrane and cytoplasm of placental trophoblast and villous trophoblast cell and cytoplasm. Analysis of all site staining results showed that in placental VCT tissue microarray, CXCR4 protein was in preeclampsia group The positive expression rate was 48.21% (27/56), the positive expression rate of the normal control group was 83.33% (35/42), and the difference between the two groups was statistically significant (U=715.5, P0.05), and the positive rate of CXCR4 protein in the preeclampsia group was 48.94% (23/47) and 79.31% (23/29) in the normal control group, and the difference was statistically significant (U=3). 62, P0.05). Conclusion miR-155 in placenta of severe preeclampsia may be involved in the pathogenesis of preeclampsia by regulating the expression of CXCR4.
【學(xué)位授予單位】:鄭州大學(xué)
【學(xué)位級(jí)別】:碩士
【學(xué)位授予年份】:2017
【分類號(hào)】:R714.244

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 許良中,,楊文濤;免疫組織化學(xué)反應(yīng)結(jié)果的判斷標(biāo)準(zhǔn)[J];中國(guó)癌癥雜志;1996年04期



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