本文選題：乙腦病毒 + 內(nèi)質(zhì)網(wǎng)應(yīng)激反應(yīng)�。� 參考：《江西農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：乙型腦炎是由乙腦病毒感染引起的中樞神經(jīng)系統(tǒng)疾病,其突出病理特征是在腦組織內(nèi)引起廣泛分布的炎癥和細(xì)胞死亡。目前針對(duì)此疾病的致病機(jī)制了解的很少,也無(wú)治療此疾病的特異性藥物。因此,研究乙腦病毒感染的表達(dá)譜變化和細(xì)胞死亡機(jī)制,進(jìn)而開(kāi)發(fā)特異性的乙型腦炎治療藥物具有重要意義。內(nèi)質(zhì)網(wǎng)應(yīng)激反應(yīng)是細(xì)胞內(nèi)外刺激引起的從內(nèi)質(zhì)網(wǎng)到胞漿和胞核的信號(hào)傳導(dǎo)途徑,該應(yīng)激反應(yīng)參與調(diào)控多種疾病,如神經(jīng)系統(tǒng)疾病、肝臟疾病,糖尿病和多種病毒性疾病等。本實(shí)驗(yàn)室的前期工作發(fā)現(xiàn)乙腦病毒可通過(guò)誘導(dǎo)內(nèi)質(zhì)網(wǎng)應(yīng)激反應(yīng)中的IRE1/JNK信號(hào)通路引起乳倉(cāng)鼠腎細(xì)胞BHK-21細(xì)胞死亡。為進(jìn)一步在蛋白質(zhì)組水平上揭示乙型腦炎致病的分子機(jī)制,本研究分析了乙腦病毒體外感染小鼠腦神經(jīng)瘤細(xì)胞Neuro2a和體內(nèi)感染鼠的腦組織的基因表達(dá)譜變化,發(fā)現(xiàn)乙腦病毒在體外和體內(nèi)均引起分子伴侶基因Hsp70表達(dá)上調(diào),誘導(dǎo)內(nèi)質(zhì)網(wǎng)應(yīng)激反應(yīng)。乙腦病毒感染也調(diào)控其他多種細(xì)胞信號(hào)通路例如FOXO、p53、TNF、、MAPK和ECM受體相關(guān)信號(hào)通路。FOXO蛋白具有廣泛的生理功能。例如調(diào)控細(xì)胞周期、衰老及凋亡。乙腦病毒感染誘導(dǎo)Neuro2a細(xì)胞凋亡,然而轉(zhuǎn)錄組測(cè)序分析、熒光定量RT-PCR和Western blot實(shí)驗(yàn)均表明,乙腦病毒感染48小時(shí),FOXO1基因以及其下游促凋亡基因Bim表達(dá)出現(xiàn)下調(diào),提示乙腦病毒誘導(dǎo)的Neuro2a細(xì)胞凋亡不是通過(guò)促凋亡基因Bim介導(dǎo)。FOXO1基因沉默可引起Neuro2a細(xì)胞核內(nèi)FOXO1和Bim轉(zhuǎn)錄水平的顯著降低,但Neuro-2a細(xì)胞凋亡率的大幅升高,與之相反的是,FOXO1的過(guò)表達(dá)造成上述二種蛋白表達(dá)量的顯著增加和Neuro-2a細(xì)胞死亡率的明顯降低。這些結(jié)果證明FOXO1可抑制Neuro-2a細(xì)胞凋亡,但不是通過(guò)調(diào)控促凋亡基因Bim。
[Abstract]:Encephalitis B is a central nervous system disease caused by encephalitis B virus infection. Little is known about the pathogenesis of the disease and no specific drug is available to treat it. Therefore, it is of great significance to study the expression profile and cell death mechanism of encephalitis B virus infection and to develop specific Japanese encephalitis drugs. Endoplasmic reticulum (ER) stress is a signal transduction pathway from endoplasmic reticulum to cytoplasm and nucleus caused by intracellular and extracellular stimuli. The stress response is involved in the regulation of many diseases, such as nervous system diseases, liver diseases, diabetes mellitus and many viral diseases. The previous work in our laboratory showed that je could induce the death of BHK-21 cells by inducing the IRE1 / JNK signaling pathway in endoplasmic reticulum stress response. In order to further reveal the molecular mechanism of encephalitis B at the proteome level, the gene expression profiles of neuro2a cells infected with Japanese encephalitis virus in vitro and the brain tissues of mice infected with Japanese encephalitis virus in vivo were analyzed. It was found that encephalitis B virus up-regulated the expression of molecular chaperone gene Hsp70 in vitro and in vivo and induced endoplasmic reticulum stress. Encephalitis B virus also regulates many other cell signaling pathways, such as FOXOp53, TNFAMAPK and ECM receptor-related signaling pathway. FOXO protein has a wide range of physiological functions. For example, regulating cell cycle, aging and apoptosis. The apoptosis of Neuro2a cells was induced by encephalitis B virus infection. However, the expression of FOXO1 gene and its downstream apoptosis-promoting gene Bim were down-regulated by transcription sequencing, fluorescent quantitative RT-PCR and Western blot assay at 48 hours after encephalitis B virus infection. The results suggested that the apoptosis induced by Japanese encephalitis virus in Neuro2a cells was not mediated by Bim-mediated. FOXO1 gene silencing induced by Japanese encephalitis virus could significantly decrease the transcription level of FOXO1 and Bim in the nucleus of Neuro2a, but the apoptosis rate of Neuro-2a cells increased significantly. In contrast, the overexpression of FOXO1 resulted in a significant increase in the expression of both proteins and a significant decrease in the cell death rate of Neuro-2a. These results suggest that FOXO1 can inhibit the apoptosis of Neuro-2a cells, but not by regulating the apoptosis-promoting gene Bim.
【學(xué)位授予單位】：江西農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R373

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 徐阿慧;郭逢林;徐晶;王騫若;張艷妮;張慶華;朱向東;賴崇德;郭韞麗;孔令保;;小鼠腦組織及培養(yǎng)神經(jīng)元Neuro-2a在內(nèi)質(zhì)網(wǎng)應(yīng)激反應(yīng)時(shí)的基因表達(dá)譜分析[J];中國(guó)生物化學(xué)與分子生物學(xué)報(bào);2017年04期

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本文編號(hào)：2101776