2011-05-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/658819摘要：葉克膜用於心肺衰竭的急救。本研究的對象為心因性休克而裝置葉克膜的病人；收集裝置葉克膜前(0小時)，裝置後2小時、72小時和移除葉克膜的四個時間點的病人血液單核細胞和血漿，研究成功與失敗兩組病人的基因體、基因表現和調節機制的主要差異，在哪個時間點出現決定性的差別。我們將以生物晶片分析成功組和失敗組的基因表現變化和microRNAs的差異，及其調控機制；尤其，分泌蛋白質基因表現的差異，可以提供作為檢測血漿中該蛋白質表現量的驗證；研究兩組病人各有哪些特殊的microRNAs變化，用以調控哪些基因，以致造成存活與失敗的差別?基因表現的調控包括遺傳性和表觀遺傳性的變化，前者為DNA序列發生改變，如有不同的單一核苷酸多型性；後者雖然具有遺傳性的傾向，但是DNA的序列是沒有改變，DNA甲基化是表觀遺傳重要的調控機制之一。兩組間單一核苷酸多型性之差別，將以單一核苷酸多型性的晶片檢測之。同時以DNA甲基化晶片研究兩組的DNA甲基化有何不同?調控機制為何?所得到的成功和失敗組之間的差異，將以生物統計方法分析，找出決定生死的時間點和相關標地，發展出預後的檢測試組，幫助臨床醫師早期判斷病人的預後；在了解調控生死的機制後，將或可進一步發展新治療法，增加ECMO病人存活率。<br> Abstract: The ECMO support maintains tissue oxygenation for days to weeks in patientswith life threatening respiratory, cardiac failure or both. Peripheral bloodmononuclear cells (PBMC) and plasma collected from cardiogenic shock patientsneeding ECMO support at 4 time points – before ECMO installed (0 hr), 2 hrs and 72hrs after installment, and ECMO removed – will be used to study genomes, geneexpression profile as well as regulation mechanisms between successful and failedpatients to find out when and what caused the difference. Microarray chips will beUsed to analyze gene expression and microRNA profiles between successful andfailed groups, differentially expressed genes and mircoRNAs will be identified andtheir regulation mechanisms will be investigated. The important secreting proteinswith differential expression will be validated by examining protein levels in plasma.What kinds of microRNAs lead to success or to failure? What target genes wereregulated by identified specific microRNAs? Regulation of gene expression occurs invarious ways, including genetic and epigenetic changes; the former refers to thechanges in the DNA sequence such as SNP polymorphism, and the later is defined asa heritable change in which the DNA sequence remains unaltered, for example, DNAmethylation. The SNP polymorphism between two groups will be examined with SNPchips and the DNA methylation differences and their regulation mechanisms betweentwo groups will also be investigated by DNA methylation chips. Once the crucialgenes and timing involved in the regulation and determinations leading to success orfailure are discovered after statistic analysis, prognosis signatures will be developedand validated; furthermore, when the regulation mechanisms are uncovered, theadvanced therapy strategies might be generated to raise ECMO survival rate andimprove patients’ life quality.微基因晶片基因表現microRNADNA甲基化葉克膜microarraygene expressionmicroRNADNA methylationECMO