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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 周崇熙 | |
| dc.contributor.author | Chia-Lan Nicole Wang | en |
| dc.contributor.author | 王嘉蘭 | zh_TW |
| dc.date.accessioned | 2021-06-15T01:49:23Z | - |
| dc.date.available | 2009-07-14 | |
| dc.date.copyright | 2009-07-14 | |
| dc.date.issued | 2009 | |
| dc.date.submitted | 2009-07-06 | |
| dc.identifier.citation | Akira, S., 2004, Toll receptor families: structure and function. Semin Immunol 16, 1-2.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/43320 | - |
| dc.description.abstract | 食用被腸炎沙門氏桿菌（Salmonella Enteritidis）污染的蛋品已經被認為是人類感染食物媒介沙門氏桿菌症主要原因之一，每年於世界各地引起許多的病例爆發，造成嚴重的經濟損失。而雞顆粒細胞（chicken granulosa cell, CGC）是排卵前濾泡中最靠近卵黃的組織層，為腸炎沙門氏桿菌入侵及增殖偏好的位置。因此本研究利用雞全基因微陣列晶片分析體外培養的雞顆粒細胞感染腸炎沙門氏桿菌後在不同時序的基因表現變化，以了解雞顆粒細胞對於感染的反應。雞顆粒細胞感染腸炎沙門氏桿菌後與未感染腸炎沙門氏桿菌的基因表現比較，共有214個基因表現量顯著地改變(p<0.01)。根據基因於感染後4小時及48小時的表現量變化，分成 1a、1b、1c、2a及2b五個子族群，分別包含70個基因(32.71%)、27個基因(12.62%)、66個基因(30.84%)、41個基因(19.16%)及10個基因(4.67%)。這些改變的基因參與免疫反應、生理調控及訊息傳遞的功能，以及部分目前未知其功能。另外，參與調控細胞激素與生長因子的 JAK-STAT pathway亦被高度活化，其中IL-6、cytokineR、STAT、PI3K四個部分有明顯變化，均屬於JAK-STAT pathway前半部。於微陣列晶片檢測結果表現量上升的基因中，與免疫反應調控相關的TLR15、IL-6、CXCLi1、CXCLi2及K203，其mRNA於兩個時間點均表現量上升，以即時定量反轉錄聚合酶連鎖反應檢測亦得到上升的結果。而與生理反應調控相關的RASD1及HB-EGF以即時定量反轉錄聚合酶連鎖反應與微陣列晶片檢測其表現量均下降。根據本實驗結果得知：雞顆粒細胞感染腸炎沙門氏桿菌後，短時間內即能夠吸引先天性免疫反應的細胞，然而長時間其免疫反應會被抑制。而且雞顆粒細胞細胞生長受到抑制，進而影響濾泡發育及排卵。其中化學激素（chemokine）及旁分泌（paracrine）扮演相當重要的角色。 | zh_TW |
| dc.description.abstract | Consumption of Salmonella Enteritidis (SE)-contaminated eggs has been recognized as one of the important reasons of human food-borne salmonellosis. Chicken granulosa cells (CGC), last tissue layer surrounding the yolk in prevulatory follicles has been demonstrated a preferred site for the Salmonella Enteritidis invasion. To understand how the chicken granulosa cell responds to the infection, a time-course in-vitro study was conducted to identify transcription changes of chicken granulosa cell using chicken whole genome microarrays. The expression of 214 genes of chicken granulosa cell had altered (p<0.01) when compared to those of uninfected cells after the infection. According to the different responses of the altered gene between 4hpi and 48hpi, they are divided into cluster 1a, 1b, 1c, 2a, and 2b those include 70 genes (32.71%), 27 genes (12.62%), 66 genes (30.84%), 41 genes (19.16%), and 10 genes (4.67%) respectively. Among of them, many were related to immune response, physiological process, signal transduction activity, transcription and some of unknown function. We also noted that the JAK-STAT pathway which is essential in regulation of cellular cytokines and growth factors being highly active those are IL-6, cytokineR, STAT,and PI3K in the upstream of this pathway. Among the up-regulated genes, the mRNA levels of TLR15, IL-6, CXCLi1, CXCLi2 and K203 at the two time points from the infected cells were also increased by the validation of real-time RT-PCR (RRT-PCR). However the mRNA expression of RASD1 and HB-EGF decreased according to the microarray and RRT-PCR analyses. These results suggest that during the Salmonella Enteritidis infection, chicken granulosa cell are capable of recruiting cells of innate immune responses in short term. In long term, the growth of chicken granulosa cell is suppressed and then the development and ovulation of follicles is also suppressed. Among of them, the chemokine and paracrine play the important role. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-15T01:49:23Z (GMT). No. of bitstreams: 1 ntu-98-R96629038-1.pdf: 38572952 bytes, checksum: f69312ff2e3f336ba45fc6fe13df65be (MD5) Previous issue date: 2009 | en |
| dc.description.tableofcontents | 中文摘要……………………………………………………………….I
英文摘要………………………………………………………………III 目錄…………………………………………………………………….V 表次………………………………………………………………......VIII 圖次…………………………………………………………………....IX 第一章 緒言………………………………………………..………….1 第二章 文獻探討………………………………………..…………….3 第一節 沙門氏桿菌及沙門氏菌症……………........………...…..3 第二節 蛋品中沙門氏桿菌污染之流行病學………..…..…...…..6 第三節 雞顆粒細胞………………………………...........…....…..8 第四節 雞細胞與沙門氏桿菌……………………........……....…10 第五節 微陣列晶片………………………. …….….........…...….18 第六節 即時定量反轉錄聚合酶鏈式反應.......…………….....…21 第三章 材料與方法…………………………………………..……....24 第一節 試劑製備……………………..........…………….….....…24 第二節 實驗雞隻……………………..........……………..…..…..25 第三節 腸炎沙門氏桿菌製備…………..........………….….……27 第四節 雞顆粒細胞製備………………….........………..….……29 第五節 腸炎沙門氏桿菌攻擊雞顆粒細胞……….……..….……30 第六節 萃取mRNA………………………………..........…..…....31 第七節 微陣列晶片及分析……………….………...........….…...32 第八節 即時定量反轉錄聚合酶鏈式反應....................................34 第四章 實驗結果………………………………….…………..….......36 第一節 mRNA品質評估…………………….…………....…...36 第二節 染劑轉換………………………………………..…..…37 第三節 雞顆粒細胞感染腸炎沙門氏桿菌後之基因表現....…38 第四節 調控路徑...............………………………………….…41 第五節 微陣列晶片與定量反轉錄聚合酶鏈式反應…............42 第五章 討論……………………………………………….….…....…44 第一節 染劑轉換（Dye swap）對微陣列晶片效能的影響....44 第二節 雞顆粒細胞之基因表現…………………….…...........46 第三節 調控路徑…………………………………….…….......50 第四節 定量反轉錄聚合酶連鎖反應與微陣列晶片效能........51 第五節 免疫相關基因表現………………………….…….......52 第六節 生理調控…………………………………….…….......60 第六章 結論………………………………………………….….........62 第七章 參考文獻……………………………………...……….......…65 表格、圖片…………………………………………...…….……........79 表 次 Table 1 The sequence of primers using in the RRT-PCR………....…79 Table 2 Mean of genes expression change-fold from M1 and M2 is more than 2-fold up-regulated at 4 hpi….…………...........80 Table 3 Mean of genes expression change-fold from M1 and M2 is more than 2-fold down-regulated at 4 hpi……...................87 Table 4 Mean of genes expression change-fold from M1 and M2 is more than 2-fold up-regulated at 48 hpi….....….................88 Table 5 Mean of genes expression change-fold from M1 and M2 is more than 2-fold down-regulated at 48 hpi…….................90 Table 6 The results of the genes expression change-fold analyzed by microarray and RRT-PCR…………….……..........................95 圖 次 Figure 1 Prehierarchal and hierarchal follicles of the laying hen…….....96 Figure 2 The follicles tissue layers surrounding the oocyte……….........97 Figure 3 The chicken granulosa cell attaches on petri dish after 24h cuture..........................................................................................98 Figure 4 The results of the RNA analyzed by Bioanalyzer 2100.............99 Figure 5 The correlation scatter diagram between M1 and M2. 5A The correlation coefficient and regression line between 4 hpi-m1 & 4 hpi-m2..........................................................101 5B The correlation coefficient and regression line between 4 hpi-m1 & 4 hpi-m2..........................................................102 Figure 6 The clusters of mRNA expression…………….…….….........103 Figure 7 The genes involved JAK-STAT pathway are altered at 4 hpi and 48 hpi…………..…...…..…………………............................104 Figure 8 The relative expression levels of genes at 4 hpi and 48 hpi analyzed by microarray and RRT-PCR...........…....……........105 | |
| dc.language.iso | zh-TW | |
| dc.subject | 基因表現 | zh_TW |
| dc.subject | 微陣列晶片 | zh_TW |
| dc.subject | 雞顆粒細胞 | zh_TW |
| dc.subject | 即時定量反轉錄聚合酶 | zh_TW |
| dc.subject | 連鎖反應 | zh_TW |
| dc.subject | gene expression | en |
| dc.subject | microarray | en |
| dc.subject | chicken granulosa cell | en |
| dc.subject | RRTPCR | en |
| dc.title | 腸炎沙門氏桿菌感染雞顆粒細胞誘發之基因時序表現研究 | zh_TW |
| dc.title | A Time-Course Study of Gene Responses in Chicken Granulosa Cells Infected by Salmonella Enteritidis | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 97-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 蔡向榮,邱智賢,張紹光,連一洋 | |
| dc.subject.keyword | 微陣列晶片,雞顆粒細胞,即時定量反轉錄聚合酶,連鎖反應,基因表現, | zh_TW |
| dc.subject.keyword | microarray,chicken granulosa cell,RRTPCR,gene expression, | en |
| dc.relation.page | 105 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2009-07-06 | |
| dc.contributor.author-college | 獸醫專業學院 | zh_TW |
| dc.contributor.author-dept | 獸醫學研究所 | zh_TW |
| 顯示於系所單位： | 獸醫學系 | |
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