母血乙肝病毒载量、胎盘Human annexin v及apoh蛋白的表达与乙肝宫内感染的相关性研究

发布时间：2018-03-17 19:32

本文选题：HBV　切入点：HAV　出处：《南昌大学》2014年硕士论文　论文类型：学位论文

【摘要】：目的：探讨孕妇血清乙肝病毒载量及足月胎盘上Human Annexin v(简称HAV)及Apolipoprotein H (简称Apoh)蛋白的表达与新生儿乙型肝炎病毒(HBV)宫内感染的相关性，，从细胞分子水平的角度来探讨HBV宫内感染发生的机制。方法：选择南昌大学第二附属医院妇产科2013年7月至2014年1月科进行产检并且至分娩的外周血HBsAg阳性（为乙肝“大三阳或小三阳”）孕妇40例作为研究组，随机选择同期于本院妇产科进行产检并且分娩的HBV正常孕妇20例作为正常对照组，并且两组孕妇的肝功能检测都为正常。根据孕妇知情同意原则，在孕妇分娩前采集肘静脉血，分娩时采集脐带血，同时无菌收集分娩时的胎盘组织，进行下研究： 1、利用PCR-荧光探针法对孕妇分娩前采集并已离心的血清中乙肝DNA的测定。 2、运用ELLESE法对孕妇血清及出生时脐血进行乙肝6项的测定。 3、通过Elivision Plus/HRP免疫组化方法检测正常孕妇及HBsAg阳性的孕妇足月胎盘中HAV及Apoh蛋白的分布及表达。结果：1、40例外周血HBsAg阳性的孕妇所生的新生儿，有3例脐血中HBsAg阳性，宫内感染率为7.5%（3/40）。 2、根据血清HBsAg阳性孕妇HBV DNA载量分3个等级，分别为104、104-107、107copies/ml，各等级组间宫内感染与宫内未感染的水平存在显著性差异（16.603，P0.05）。 3、免疫组化结果示Annexin v及Apoh蛋白在人胎盘各层细胞中均有表达，其主要表达于滋养细胞(TC)，绒毛间质细胞(VMC)和绒毛毛细血管内皮细胞(VCMC)，且从胎盘的母体面到胎儿面Annexin v及Apoh蛋白阳性信号逐渐呈现下降的趋势。 4、血HBsAg阳性孕妇足月胎盘组织中HAV及Apoh蛋白的表达明显高于对照组(P0.05），且HAV及Apoh蛋白在HBVDNA阳性组胎盘中的表达明显高于HBVDNA阴性组胎盘中表达(P0.05）。HAV及Apoh蛋白在外周血HBsAg阳性孕妇足月胎盘组织中的表达呈正相关性（r=0.538，P0.05）。结论：1、孕妇外周血HBVDNA载量为乙肝宫内感染的危险因素之一，且随着孕产妇血HBV-DNA载量的增加，胎儿宫内HBV感染的危险性呈上升趋势。 2、 HAV、Apoh蛋白广泛表达于胎盘各层的各型细胞内，且从母体面到胎儿面蛋白表达率阳性表达率呈逐渐下降的趋势。 3、HAV、Apoh蛋白在HBsAg阳性胎盘组织中的表达明显上调，且两者呈正相关，两者在乙肝宫内感染的发生、发展中起协同作用。
[Abstract]:Objective: to investigate the relationship between the serum hepatitis B virus load and the expression of Human Annexin v and Apolipoprotein H protein in the placenta of pregnant women and the intrauterine infection of neonatal hepatitis B virus (HBV). To explore the mechanism of intrauterine infection of HBV at cellular and molecular level. Methods: from July 2013 to January 2014, 40 pregnant women with HBsAg positive in peripheral blood of the second affiliated Hospital of Nanchang University, Department of Obstetrics and Gynecology and Obstetrics and Gynecology and Obstetrics, were selected as the study group. Twenty normal pregnant women with HBV were randomly selected as normal control group. According to the principle of informed consent of pregnant women, the blood samples of cubital vein were collected before delivery. Umbilical cord blood was collected during delivery and placental tissue during delivery was collected aseptically. The following studies were carried out:. 1. PCR- fluorescence probe method was used to detect HBV DNA in serum collected before delivery and centrifuged by pregnant women. 2. ELLESE method was used to detect 6 items of hepatitis B in pregnant women's serum and umbilical cord blood at birth. 3. The distribution and expression of HAV and Apoh protein in the placenta of normal and HBsAg positive pregnant women were detected by Elivision Plus/HRP immunohistochemical method. Results of 40 newborns born to 40 pregnant women with HBsAg positive peripheral blood, 3 were HBsAg positive in umbilical cord blood. The intrauterine infection rate was 7.5 / 40. 2. According to the HBV DNA load of serum HBsAg positive pregnant women, there were three grades of HBV DNA loading, which were 104 ~ 104-107 ~ 107 / ml, respectively. There was significant difference between intrauterine infection and intrauterine infection among different grades (16.603g / ml, P = 0.05). 3Immunohistochemical results showed that Annexin v and Apoh protein were expressed in human placental cells. It was mainly expressed in trophoblast, villous stromal cell (VMC) and chorionic capillary endothelial cell (VCM C), and the positive signals of Annexin v and Apoh protein decreased gradually from the maternal surface of placenta to the fetal surface. 4. The expression of HAV and Apoh protein in placenta of pregnant women with positive HBsAg was significantly higher than that of control group (P0.05A), and the expression of HAV and Apoh protein in placenta of HBVDNA positive group was significantly higher than that of HBVDNA negative group and Apoh protein in peripheral blood HBsAg. There was a positive correlation between the expression of the positive pregnant women and the placental tissues of the full-term pregnant women (P 0.05). Conclusion the HBVDNA load in maternal peripheral blood is one of the risk factors of HBV intrauterine infection, and the risk of fetal intrauterine HBV infection increases with the increase of maternal blood HBV-DNA load. (2) HAV Apoh protein was widely expressed in all kinds of cells of placenta, and the positive expression rate of HAV Apoh protein decreased gradually from maternal surface to fetal surface. (3) the expression of Apoh protein in HBsAg positive placenta was significantly up-regulated, and there was a positive correlation between them. The two proteins played a synergistic role in the occurrence and development of HBV intrauterine infection.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R714.251

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