Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/12790
標題: S. choleraesuis與S. typhimurium在巨噬細胞內存活能力及誘導細胞計 畫性死亡之比較
Comparisons of S. choleraesuis and S. typhimurium on surviving and inducing apoptosis in infected macrophage.
作者: 廖志明
Laiw, Jyh-ming
關鍵字: Salmonelosis;沙氏桿菌症;intracellular bacteria;intracellular surviving;apoptosis;細胞內寄生性細菌;細胞內存活;細胞計畫性死亡
出版社: 獸醫學系
摘要: 
摘 要 S. choleraesuis與S. typhimurium為導致豬隻沙氏
桿菌症之主要病原,常造成被感染宿主下痢與敗血症。因為沙氏桿菌之血
清型眾多, 而傳統鑑定方法頗為複雜又費時,因此,利用隨機增幅聚合
酵素連鎖反應(RAPD- PCR)建立分子鑑定系統不失為一方便快速且準確之
鑑別 方法,本試驗由60組隨機引子所挑選出之E04及J06二組引子在42℃
的煉合溫度下,可精確與快速的來區分S. choleraesuis及S.
typhimurium 並具有高度之區分性及再現性。沙氏桿菌已被證實是兼性
細胞內寄生性細菌,具有在吞噬細胞中存活的能力,為深入探討此二種沙
氏桿菌 分別造成豬隻下痢或敗血症之機制,是否與其在吞噬細胞中存活
能力的差異有關,本試驗應用流體細胞儀技術,檢測S. choleraesuis及
S.typhimurium於吞噬細胞株RAW264.7與SPF豬肺泡巨噬細胞內之存活能力
。結果中發現S. choleraesuis不論在吞噬細胞株RAW264.7或SPF 豬隻肺
泡巨噬細胞內均比S. typhimurium更具有較佳的存活能力。此外,沙氏桿
菌在感染過程中必須面對消化道中多種不同的酸性環境, 本試驗亦對細
菌以酸性環境處理,探討其對細菌在吞噬細胞中的存活能力之影響,結果
顯示經不同程度之酸性環境適應後,二種沙氏桿菌 皆可明顯提高其在不
同吞噬細胞中之存活能力。至於在感染吞噬細胞後之作用機制上,沙氏桿
菌不僅能躲避吞噬細胞內之毒殺作用而能 存活下來,最後並能導致吞噬
細胞的細胞計畫性死亡(apoptosis),本實驗亦分別利用Annexin-V、
TUNEL assay及核酸階梯現象,證實 在不同的感作時間S.choleraesuis
及S. typhimurium皆能誘導吞噬細胞產生細胞之計畫性死亡現象,且S.
choleraesuis在肺泡巨噬細胞 所造成的apoptosis亦較S. typhimurium
明顯。

Summary The diarrheagenic disease Salmonellosis is
among the most important bacterial diseases affecting swine
worldwide and the causative agents S. choleraesuis and S.
typhimurium also can induce gastroenteritis and systemic
septicemia in human. The random amplified polymorphic DNA
(RAPD) PCR was utilized to distinguish fingerprinting profiles
of S. choleraesuis and S. typhimurium. A total of 60 random
primers were applied and among them, E4 and J6 primers with
annealing temperature of 42C exhibited better accuracy for
identification. To investigate the pathogenesis of Salmonella
after infection, murine macrophage cell line (RAW264.7) and
alveolar macrophage from SPF swine were used for comparisons the
surviving of S. choleraesuis and S. typhimurium in infected
cells by flow cytometry analysis. The results demonstrated that
S. choleraesuis showed higher survival abilities in both types
of macrophages. Moreover, S. choleraesuis and S. typhimurium
adapted in low pH environment before internalizing with
macrophages both could enhance their durability against
macrophages killing. In addition,Annexin-V, TUNEL assay and DNA
fragmentation were also utilized to detect programmed cell death
of macrophages. The results indicated both S. choleraesuis and
S. typhimurium could induce variety of apoptosis after
subsequently invasion; however, S. choleraesuis provoked
stronger responses of apoptosis and cytotoxicity in alveolar
macrophages.
URI: http://hdl.handle.net/11455/12790
Appears in Collections:獸醫學系所

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