新生与成年土拨鼠肝组织中部分差异表达的基因比较分析

发布时间：2018-01-21 05:29

本文关键词： 基因芯片土拨鼠钙离子土拨鼠肝炎病毒　出处：《北京协和医学院》2013年硕士论文　论文类型：学位论文

【摘要】：目前,乙型肝炎病毒(Hepatitis B virus, HBV)仍以其庞大的人口基数威胁着人类的健康和生命。新生儿感染HBV,大部分发展为慢性肝炎,而成年人感染HBV则多发生急性自限性肝炎。HBV导致的慢性肝炎进而发生肝纤维化、肝硬化,最终以肝癌结局,而急性自限性肝炎多以自愈结局。很多科研工作者研究了慢性肝炎和急性自限性肝炎的感染和发病过程,试图寻找HBV在两个年龄段引发不同病症的原因,但未取得太多成果。新生和成年土拨鼠感染土拨鼠肝炎病毒(woodchuck hepatitis virus, WHV)后,发病状况与对应年龄人类感染HBV后相似。本课题拟用土拨鼠寻找肝组织中可能导致这种预后差异的关键基因。采用全基因组表达谱芯片技术,对比新生与成年小鼠肝组织差异表达基因,选取目的基因,再通过多个物种序列比对,设计简并引物,在土拨鼠肝组织cDNA中扩增对应基因片段,测序,再次设计引物,进行实时荧光定量PCR。与新生士拨鼠相比,成年土拨鼠肝细胞中与钙离子重吸收相关基因Dnml (Dynamin1)、Dnm3(Dynamin3)及Prkcc (Protein kinase C, gamma)表达率明显升高,分别上升2.65+0.25倍,1.90±0.34倍,2.94-0.54倍。钙离子重吸收通路中,Dnm1、Dnm3、Prkcc在两组动物肝脏中表达差异最明显,并且在新生组与成年组土拨鼠之间也有明显差异。此类基因造成肝细胞内钙离子浓度的差别,间接影响其中肝炎病毒的复制。这种表达差异很可能是导致两个年龄段动物感染土拨鼠肝炎病毒后转归不同的原因之一。
[Abstract]:At present, hepatitis B virus (HBV) is still threatening human health and life with its huge population base. The newborn is infected with HBV. Most of them develop into chronic hepatitis, while adults infected with HBV often develop acute self-limited hepatitis. HBV-induced chronic hepatitis and liver fibrosis, liver cirrhosis, and finally liver cancer. Many researchers have studied the infection and pathogenesis of chronic hepatitis and acute self-limited hepatitis, and tried to find the causes of HBV in the two age groups. But not much. New and adult groundhog infected with woodchuck hepatitis virus (WHV). The incidence of the disease is similar to that in humans of the corresponding age after infection with HBV. This study intends to use groundhog to search for key genes in liver tissue that may lead to this difference in prognosis. The differentially expressed genes in the liver tissues of newborn and adult mice were compared by using the whole genome expression microarray technique. The target genes were selected and then degenerate primers were designed by multiple species sequence alignment. The corresponding gene fragments were amplified in cDNA of groundhog liver, sequenced, primers were redesigned and real-time fluorescence quantitative PCR was carried out. In adult groundhog liver cells, the calcium reabsorption related gene Dnml / Dynamin 1 was found. The expression rates of Prkcc protein kinase C, gamma were increased by 2.65 0.25 times, respectively. 1.90 卤0.34 times, 2.94-0.54 times. The expression of Dnm1 and Dnm3 Prkcc in liver of the two groups was the most significant difference in calcium reabsorption pathway. There were also significant differences between the newborn and adult groundhog groups. The difference of calcium concentration in hepatocytes was caused by these genes. The difference in expression of hepatitis virus may be one of the reasons why the two age groups are infected with groundhog hepatitis virus.
【学位授予单位】：北京协和医学院
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R512.62

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