https://scholars.lib.ntu.edu.tw/handle/123456789/192317
Title: | 以基因微陣列分析肝細胞再生時,特種基因及基因族群之基因表現在程度、型態、時程的變遷並鑑別其扮演之角色(2/2) ANALYZING THE REGULATING GENES OF LIVER REGENERATION IN CHANGING DEGREE, PATTERN, TIMING AND VERIFYING THE ROLES OF SPECIFIC AND CLUSTER GENES BY cDNA MICROARRY (2/2) |
Authors: | 賴鴻緒 | Keywords: | 肝細胞再生;部分肝臟切除術;proto-oncogene;基因微陣列;基因表現型 態;基因族群;liver regeneration;partial hepatectomy;microarray;genetic changing pattern;gene cluster | Issue Date: | 31-Jul-2005 | Publisher: | 臺北市:國立臺灣大學醫學院外科 | Abstract: | 有關肝細胞在肝損傷後可以再生,雖 已是公認的事實,許多研究也證實多種營 養素、賀爾蒙、生長因子、藥劑等,可直 接或間接影響肝細胞再生,但肝細胞再生 基因控制之詳細機轉,則仍不明瞭。 本計畫第一年以大鼠切除70%肝臟 模式研究發現:(1)剩餘肝臟重量比值,於 切肝術後72 小時即恢復90%以上;(2)有絲 分裂於術後48 小時大量出現,術後72 小 時逐漸減少;(3)所有基因表現變遷之型態 及時程共分為72 種,包括第2、、6、12、 24、72 小時及、7 天單一尖峰型、雙尖峰、 遞增型、遞減型、突出型、凹陷型和混和 型等,每種型態包括40 至218 種基因;(4) 包括免疫、賀爾蒙、生長因子、酵素及血 管新生因子等基因族群,均有明顯的變 遷;(5) fas-associating protein with death domain, carnitine palmitoyltransferase 1, fas death domain-associating protein, 及steroid O-acyltransferase 1 等早期變化基因可能與 肝再生之啟動有關;(6) transforming growth factor beta 2 及beta receptor 等中期變化基 因可能與肝再生之分化有關;(7) TGF-β regulated gene 3 及small inducible cytokine A2 等晚期變化基因可能與肝再生之終止 有關。進一步以切除較少(40%)肝臟比較其 程度差異,以及比對各種基因之重要性, 則於第二年進行。 本計劃第二年仍以重約200 克之 Wistar 雄性大鼠做實驗,測定肝細胞再生 過程中,超過20000 種基因種別之基因表 現,並分析及比較上一年70%切肝後,其 變遷程度、型態、時程及各基因族群之相 關性。所有大鼠均接受約百分之四十之肝 臟部分切除手術,各於術前及術後2、4、6、 12、24、72 小時及5、7、10 天後犧牲取樣, 仍測定:(1)剩餘肝臟之重量比值;(2)剩餘 肝臟之有絲分裂指標;(3)以基因微陣列高 密度晶片、肝細胞mRNA 標號、 hybridization 及影像分析等方法,測定 20,500 種基因表現之變遷程度、型態及時 程;(4)將基因表現變化大者,依特性分為 免疫、賀爾蒙、生長因素、酵素、及血管 新生因子等基因族群,並比較其程度、型 態及時程與70%切肝時,變化之差異性。 結果發現:(1)剩餘肝臟重量比值,切肝40 %後恢復速度較慢,但至術後72 小時,40 %與70%兩組幾乎到達同樣重量;(2)有絲分裂切肝40%後48 小時也出現,至72 小 時逐漸減少,但程度上比70%切肝後明顯 較少;(3)基因表現變遷之種類與型態,與 70%切肝組類似,亦為72 種型態,部分基 因變化程度雖較低,但變化型態相近;(4) 包括免疫、賀爾蒙、生長因子、酵素及血 管新生因子等基因族群,均有明顯的變 遷;(5)免疫相關基因(如IL-6 和IL-10)及血 管新生相關基因( 如Angiotensiongen, VEGF, 及VEGF receptor 2)之變化40%與 70%切肝後相近於重量比值變化,可能在 肝細胞再生之調控,扮演較重要的角色。 Although there are much controversy on the initiation, regulation, metabolic changes, and termination of liver regeneration after partial hepatectomy that well initiate proliferation of the remaining hepatocytes, several factors, such as hormones, growth factors, nutritional components, and pharmacological agents, have been demonstrated to directly or indirectly affect liver regeneration. However, the regenerative mechanism and genetic control of liver after major tissue loss is still not clear. We used 70% portal hepatectomized rat model for studying genes variation after partial hepatectomy, and found that: (1) the remnant liver weight increased to 90% in 72h after partial hepatectomy; (2) the mitosis of hepatocytes increased marked at 48h then decreased at 72h after partial hepatectomy; (3) analyzing the gene expression of microarray chips, the variation could be classified into 72 different patterns in cluding the patterns with a single peak at 2, 4, 6, 12, 24, 72h and 5, 7d after partial hepatectomy; (4) gene clusters of immune, hormone, growth factor, enzyme and angiogenesis have changed markedly; (5) early stage changed genes including fas-associating protein with death domain, carnitine palmitoyltransferase 1, fas death domain-associating protein, and steroid O-acyltransferase 1 could be related to the initiation of liver regeneration; (6) intermediate stage changed genes including transforming growth factor beta 2 and beta receptor could be related to the differentiation of liver regeneration; (7) late stage changed genes including TGF-β regulated gene 3 and small inducible cytokine A2 could be related to the termination of liver regeneration. Study with 40% partial hepatectomy was performed in the second year. Male Wistar rats around 200g will be used as subject. Partial hepatectomy around 40% were performed. They were sacrificed before and 2, 4, 6, 12, 24, 72 hours and 5, 7, 10 days after hepatectomy. We have measured: (1)weight of remnant liver; (2)mitotic index; (3)genomic survey of the gene expression by microarray chip of 20,500 identified cDNA clones, labeling of liver mRNA hybridization and image analysis; and (4)Grouping of genes expression into immune, nutrition, hormone, growth factor, enzyme, oncologic and embryonic subgroups, and compare the expression degree, changing pattern and specific timing. The results were: (1) the remnant liver weight recovered slower, but can reach 90% in 72h after partial hepatectomy; (2) the mitosis of hepatocytes also increased markedly at 48h although not so high as 70% group rats, and also decreased at 72h after partial hepatectomy; (3) analyzing the gene expression of microarray chips, the variation of 40% group rats could also be classified into 72 patterns just like 70% group, with some variation degree were not so marked as 70% group rats; (4) gene clusters of immune, hormone, growth factor, enzyme and angiogenesis have changed markedly; (5) The changes of gene expression in immune related genes (such as IL-6 and IL-10 control genes) and angiogenesis related genes (such as Angiotensiongen, VEGF, and VEGF-receptor 2) were more similar to the remnant liver weight variations. It may indicate that these genes play more important roles in the control mechanism of liver regeneration. |
URI: | http://ntur.lib.ntu.edu.tw//handle/246246/24545 | Other Identifiers: | 932314B002056 | Rights: | 國立臺灣大學醫學院外科 |
Appears in Collections: | 醫學系 |
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