本文關(guān)鍵詞：流感病毒感染小鼠肺組織中p53靶基因鑒定　出處：《中國農(nóng)業(yè)科學(xué)院》2014年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 流感病毒 炎癥反應(yīng) NF-κB Nfkbiz p53

【摘要】：流感病毒感染能夠誘發(fā)機(jī)體炎癥反應(yīng),而人類歷史上的幾次引起流感的流行與暴發(fā)的流感病毒均能引起強(qiáng)烈的炎癥反應(yīng),所引起細(xì)胞因子風(fēng)暴造成多種組織和器官的嚴(yán)重?fù)p傷。尤其是肺泡巨噬細(xì)胞感染病毒后釋放大量的促炎性細(xì)胞因子引起的炎性病理損傷,如廣泛的肺水腫、肺泡出血組織損傷和壞死,使機(jī)體發(fā)生多器官的功能衰竭,成為引起流感患者死亡的主要原因。NF-κB作為炎癥反應(yīng)的關(guān)鍵調(diào)控分子,能夠促進(jìn)多種促炎性細(xì)胞因子的合成與釋放,促進(jìn)炎癥反應(yīng)的發(fā)生。NF-κB的激活常被認(rèn)為是炎癥反應(yīng)起始的標(biāo)志。腫瘤抑制因子p53在受到多種應(yīng)激條件的刺激下激活,不僅在DNA損傷修復(fù)、細(xì)胞周期停滯、細(xì)胞凋亡中以及抗病毒感染中發(fā)揮作用,還具有抑制炎癥反應(yīng)的效應(yīng)。許多研究發(fā)現(xiàn),NF-κB與p53這兩大信號(hào)通路之間在許多生理病理?xiàng)l件下都存在著相互作用,如免疫系統(tǒng)的發(fā)育、新陳代謝、細(xì)胞凋亡、腫瘤發(fā)生以及炎癥反應(yīng)。p53與NF-κB在流感病毒感染中誘發(fā)的炎癥反應(yīng)中又有著怎樣的相互作用,p53是否在其中扮演者抑制NF-κB的活性而抑制炎癥反應(yīng)的角色。 本研究首先用流感病毒感染p53野生型C57BL/6小鼠后,對(duì)采取的肺組織進(jìn)行破碎后進(jìn)行ChIP-seq測(cè)序以發(fā)現(xiàn)轉(zhuǎn)錄因子p53潛在的靶基因,繼而發(fā)現(xiàn)p53潛在調(diào)控的靶基因中有與炎癥反應(yīng)相關(guān)的基因。這表明p53作為轉(zhuǎn)錄調(diào)控因子,在炎癥反應(yīng)的發(fā)生中可能通過轉(zhuǎn)錄調(diào)控炎癥相關(guān)分子的基因從而抑制炎癥反應(yīng)。分析ChIP-seq的結(jié)果發(fā)現(xiàn),炎癥反應(yīng)通路中的Nfkbiz基因可能為p53潛在的靶基因。 Nfkbiz基因編碼的IκB(?)屬于IKB家族成員,在NF-κB信號(hào)通路中屬于非典型成員,能夠直接在細(xì)胞核內(nèi)與NF-κB二聚體結(jié)合影響NF-κB的轉(zhuǎn)錄活性。研究發(fā)現(xiàn),IκB(?)屬于IKB家族成員,不僅具有抑制NF-κB的轉(zhuǎn)錄活性,同時(shí)也是NF-κI某些下游靶基因轉(zhuǎn)錄所必須的組成成分,特別是在NF-κB引起的炎癥反應(yīng)第二期反應(yīng)中細(xì)胞因子所對(duì)應(yīng)的基因.IκB(?)有可能在流感病毒感染中扮演著通過調(diào)控NF-κB的轉(zhuǎn)錄活性進(jìn)而調(diào)節(jié)炎癥反應(yīng)的作用。 通過熒光實(shí)時(shí)定量方法,我們發(fā)現(xiàn)過表達(dá)p53能夠抑制Nfkbiz基因的轉(zhuǎn)錄水平,在使用LPS刺激Nfkbiz基因表達(dá)的同時(shí)過表達(dá)p53發(fā)現(xiàn)抑制效應(yīng)更加明顯,表明p53可能通過抑制Nfkbiz基因的轉(zhuǎn)錄從而抑制過度的炎癥反應(yīng)。對(duì)Nfkbiz基因序列進(jìn)行分析后,發(fā)現(xiàn)了其中含有可能的p53反應(yīng)元件。繼而通過構(gòu)建的熒光素酶報(bào)告質(zhì)粒進(jìn)行熒光素酶雙報(bào)告試驗(yàn),發(fā)現(xiàn)p53確實(shí)能夠與Nfkbiz基因中的p53反應(yīng)元件結(jié)合并抑制Nfkbiz基因的轉(zhuǎn)錄。ChIP-PCR和ChIP-QPCR的結(jié)果也表明p53能夠與Nfkbiz基因中+8947～+8969序列進(jìn)行結(jié)合。以上結(jié)果表明Nfkbiz是p53的靶基因,能夠被p53抑制轉(zhuǎn)錄活性。 綜上所述,本研究證明Nfkbiz是p53負(fù)向調(diào)控的靶基因,p53能夠通過抑制Nfkbiz編碼IκB(?),從而降低NF-κB在流感病毒感染中的誘發(fā)炎癥反應(yīng)的水平。本研究發(fā)現(xiàn)了p53在抑制炎癥反應(yīng)中可能存在的新機(jī)制,拓展了p53的細(xì)胞分子生物學(xué)作用,為研究p53在流感病毒感染過程中能夠降低死亡率提供了新的角度。
[Abstract]:NF - 魏B acts as a key regulator of inflammatory response , and can promote the synthesis and release of various cytokines . NF - 魏B can promote the synthesis and release of inflammatory cytokines . NF - 魏B can promote the synthesis and release of inflammatory cytokines . In this study , we first infected p53 wild - type C57BL / 6 mice with influenza virus , and then sequenced the lung tissues to find the potential target gene of transcription factor p53 . It is found that I魏B ( ? ) is a member of IKB family , which can directly bind NF - 魏B dimer to NF - 魏B transcription activity . It is found that I魏B ( ? ) is a member of IKB family , which not only has the transcription activity to inhibit NF - 魏B , but also the gene involved in the transcription of some downstream target genes of NF - 魏B , especially in the second phase of inflammatory response induced by NF - 魏B ( ? ) . I魏B ( ? ) may play a role in regulating the transcriptional activity of NF - 魏B and regulating inflammatory response . It was found that p53 was able to inhibit the transcription of Nfk2gene . The results of ChIP - PCR and ChIP - QPCR also showed that p53 was able to bind to and inhibit the transcription of Nfkbizgene . The results showed that p53 was the target gene of p53 , which was able to inhibit the transcription activity of p53 . In conclusion , the present study proves that Nfkbiza is a target gene for p53 negative regulation , and p53 can inhibit the level of inflammatory response induced by NF - 魏B in influenza virus infection .

【學(xué)位授予單位】：中國農(nóng)業(yè)科學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：S852.65;R373

【共引文獻(xiàn)】

相關(guān)期刊論文 前10條

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3 Hedi Harizi;Norbert Gualde;;Pivotal Role of PGE_2 and IL-10 in the Cross-Regulation of Dendritic Cell-Derived Inflammatory Mediators[J];Cellular & Molecular Immunology;2006年04期

4 李泉;俞衛(wèi)鋒;邱必軍;張金e，

本文編號(hào)：1359501