PI4KA与乙型肝炎病毒复制的相关性研究

发布时间：2018-05-08 00:16

  本文选题：肝炎病毒 + 肝炎　； 参考：《青岛大学》2014年硕士论文

【摘要】：目的：通过探讨PI4KA与慢性乙型肝炎患者病毒载量的关系；观察PI4KA在肝癌细胞株HepG2及HepG2.2.15中表达的差异,探究HBV对PI4KA表达的影响;在临床和细胞水平探讨PI4KA与乙型肝炎病毒复制的关系。方法：收集180例慢性乙型肝炎患者的血清,利用荧光定量PCR检测其HBV DNA的载量,根据其HBV DNA载量的高低,分为三组：高病毒载量组(HBV DNA1×107拷贝/m1)、中病毒载量组(1×105拷贝/ml≤HBV DNA≤1×107拷贝/m1)及低病毒载量组(]3BV DNA1×105拷贝/m1),同时以60名健康人作对照。采用ELISA法,检测各组外周血清中的PI4KA的含量；培养HepG2及HepG2.2.15细胞,当细胞融合达到90%时,提取两细胞RNA并逆转录成cDNA,采用实时荧光定量PCR法检测HepG2和HepG2.2.15细胞中PI4KA mRNA的表达水平；采用ELISA法检测此两种细胞培养上清液中PI4KA的表达水平。 结果：60例健康人血清PI4KA的含量为2.29±0.75ng/ml;60例低病毒载量组血清PI4KA的含量为2.73±0.71ng/ml;60例中病毒载量组血清PI4KA的含量为3.52±0.78ng/ml及60例高病毒载量组血清PI4KA含量为4.72±0.77ng/ml。慢性乙型肝炎患者不同病毒载量组与健康对照组间血清PI4KA的含量差异有显著统计学意义(F=119.958,P0.01),并且不同病毒载量组都显著高于对照组。随着HBV DNA载量的升高,PI4KA的含量增加,两者呈正相关(r=0.758, P0.01); HepG2.2.15细胞中PI4KA mRNA的表达水平较HepG2高,差异有统计学意义(P0.05)；在细胞培养上清液中,HepG2和HepG2.2.15细胞PI4KA的含量分别为1.07±0.54ng/ml和1.63±0.43ng/ml, HepG2.2.15细胞中PI4KA的含量高于HepG2细胞(-2.29,P0.05)。 结论：慢性乙型肝炎患者血清PI4KA的含量与HBV DNA载量密切相关；HBV能够在mRNA和蛋白水平上调肝癌细胞PI4KA的表达；在临床水平和细胞水平证实PI4KA与乙型肝炎病毒复制密切相关,乙型肝炎病毒可能通过上调宿主因子PI4KA的表达来促进自身病毒的复制。
[Abstract]:Objective: to investigate the relationship between PI4KA and viral load in patients with chronic hepatitis B, to observe the difference of PI4KA expression in HepG2 and HepG2.2.15, and to explore the effect of HBV on PI4KA expression. To investigate the relationship between PI4KA and hepatitis B virus replication at the clinical and cellular levels. Methods: the serum samples of 180 patients with chronic hepatitis B were collected and their HBV DNA load was detected by fluorescence quantitative PCR, and the HBV DNA load was measured according to the level of HBV DNA load. They were divided into three groups: high viral load group (DNA1 脳 10 ~ 7 copy / m ~ (1), medium viral load group (n = 1 脳 10 ~ 5 copies / ml 鈮，

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