發(fā)布時間：2018-03-17 19:32

  本文選題：HBV　切入點：HAV　出處：《南昌大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：探討孕婦血清乙肝病毒載量及足月胎盤上Human Annexin v(簡稱HAV)及Apolipoprotein H (簡稱Apoh)蛋白的表達與新生兒乙型肝炎病毒(HBV)宮內(nèi)感染的相關(guān)性，，從細胞分子水平的角度來探討HBV宮內(nèi)感染發(fā)生的機制。 方法：選擇南昌大學(xué)第二附屬醫(yī)院婦產(chǎn)科2013年7月至2014年1月科進行產(chǎn)檢并且至分娩的外周血HBsAg陽性（為乙肝“大三陽或小三陽”）孕婦40例作為研究組，隨機選擇同期于本院婦產(chǎn)科進行產(chǎn)檢并且分娩的HBV正常孕婦20例作為正常對照組，并且兩組孕婦的肝功能檢測都為正常。根據(jù)孕婦知情同意原則，在孕婦分娩前采集肘靜脈血，分娩時采集臍帶血，同時無菌收集分娩時的胎盤組織，進行下研究： 1、利用PCR-熒光探針法對孕婦分娩前采集并已離心的血清中乙肝DNA的測定。 2、運用ELLESE法對孕婦血清及出生時臍血進行乙肝6項的測定。 3、通過Elivision Plus/HRP免疫組化方法檢測正常孕婦及HBsAg陽性的孕婦足月胎盤中HAV及Apoh蛋白的分布及表達。 結(jié)果：1、40例外周血HBsAg陽性的孕婦所生的新生兒，有3例臍血中HBsAg陽性，宮內(nèi)感染率為7.5%（3/40）。 2、根據(jù)血清HBsAg陽性孕婦HBV DNA載量分3個等級，分別為104、104-107、107copies/ml，各等級組間宮內(nèi)感染與宮內(nèi)未感染的水平存在顯著性差異（16.603，P0.05）。 3、免疫組化結(jié)果示Annexin v及Apoh蛋白在人胎盤各層細胞中均有表達，其主要表達于滋養(yǎng)細胞(TC)，絨毛間質(zhì)細胞(VMC)和絨毛毛細血管內(nèi)皮細胞(VCMC)，且從胎盤的母體面到胎兒面Annexin v及Apoh蛋白陽性信號逐漸呈現(xiàn)下降的趨勢。 4、血HBsAg陽性孕婦足月胎盤組織中HAV及Apoh蛋白的表達明顯高于對照組(P0.05），且HAV及Apoh蛋白在HBVDNA陽性組胎盤中的表達明顯高于HBVDNA陰性組胎盤中表達(P0.05）。HAV及Apoh蛋白在外周血HBsAg陽性孕婦足月胎盤組織中的表達呈正相關(guān)性（r=0.538，P0.05）。 結(jié)論：1、孕婦外周血HBVDNA載量為乙肝宮內(nèi)感染的危險因素之一，且隨著孕產(chǎn)婦血HBV-DNA載量的增加，胎兒宮內(nèi)HBV感染的危險性呈上升趨勢。 2、 HAV、Apoh蛋白廣泛表達于胎盤各層的各型細胞內(nèi)，且從母體面到胎兒面蛋白表達率陽性表達率呈逐漸下降的趨勢。 3、HAV、Apoh蛋白在HBsAg陽性胎盤組織中的表達明顯上調(diào)，且兩者呈正相關(guān)，兩者在乙肝宮內(nèi)感染的發(fā)生、發(fā)展中起協(xié)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.
【學(xué)位授予單位】：南昌大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R714.251

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