本文關(guān)鍵詞： 基因芯片 土撥鼠 鈣離子 土撥鼠肝炎病毒　出處：《北京協(xié)和醫(yī)學(xué)院》2013年碩士論文　論文類型：學(xué)位論文

【摘要】：目前,乙型肝炎病毒(Hepatitis B virus, HBV)仍以其龐大的人口基數(shù)威脅著人類的健康和生命。新生兒感染HBV,大部分發(fā)展為慢性肝炎,而成年人感染HBV則多發(fā)生急性自限性肝炎。HBV導(dǎo)致的慢性肝炎進而發(fā)生肝纖維化、肝硬化,最終以肝癌結(jié)局,而急性自限性肝炎多以自愈結(jié)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發(fā)病過程,試圖尋找HBV在兩個年齡段引發(fā)不同病癥的原因,但未取得太多成果。新生和成年土撥鼠感染土撥鼠肝炎病毒(woodchuck hepatitis virus, WHV)后,發(fā)病狀況與對應(yīng)年齡人類感染HBV后相似。本課題擬用土撥鼠尋找肝組織中可能導(dǎo)致這種預(yù)后差異的關(guān)鍵基因。 采用全基因組表達譜芯片技術(shù),對比新生與成年小鼠肝組織差異表達基因,選取目的基因,再通過多個物種序列比對,設(shè)計簡并引物,在土撥鼠肝組織cDNA中擴增對應(yīng)基因片段,測序,再次設(shè)計引物,進行實時熒光定量PCR。與新生士撥鼠相比,成年土撥鼠肝細胞中與鈣離子重吸收相關(guān)基因Dnml (Dynamin1)、Dnm3(Dynamin3)及Prkcc (Protein kinase C, gamma)表達率明顯升高,分別上升2.65+0.25倍,1.90±0.34倍,2.94-0.54倍。 鈣離子重吸收通路中,Dnm1、Dnm3、Prkcc在兩組動物肝臟中表達差異最明顯,并且在新生組與成年組土撥鼠之間也有明顯差異。此類基因造成肝細胞內(nèi)鈣離子濃度的差別,間接影響其中肝炎病毒的復(fù)制。這種表達差異很可能是導(dǎo)致兩個年齡段動物感染土撥鼠肝炎病毒后轉(zhuǎn)歸不同的原因之一。
[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 鹵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.
【學(xué)位授予單位】：北京協(xié)和醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R512.62

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相關(guān)期刊論文 前1條

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本文編號：1450606