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研究生:李宥萱
研究生(外文):Yu-hsuan Lee
論文名稱:戴奧辛與五氯酚誘發免疫毒性之可能的協同作用影響探討
論文名稱(外文):Study on the possible synergistic effect of TCDD and Pentachlorophenol on immunotoxicity
指導教授:王應然王應然引用關係
指導教授(外文):Ying-Jan Wang
學位類別:碩士
校院名稱:國立成功大學
系所名稱:環境醫學研究所
學門:醫藥衛生學門
學類:公共衛生學類
論文種類:學術論文
論文出版年:2008
畢業學年度:96
語文別:中文
論文頁數:73
中文關鍵詞:BALB/c小鼠免疫抑制作用戴奧辛五氯酚雞卵白蛋白(OVA)
外文關鍵詞:BALB/c micePentachlorophenolImmune suppressionTCDDAlbumin(OVA)
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五氯酚(Pentachlorophenol, PCP)與戴奧辛(dioxin)為環境中著名的污染物質。工業上常用的PCP多為含多氯戴奧辛的不純物。這些不純的含氯化合物被懷疑為導致PCP具有免疫毒性能力的關鍵物質。戴奧辛中以2,3,7,8-四氯戴奧辛(2,3,7,8-Tetrachlorodibenzo-p-dioxin ,TCDD)毒性最強,且對於免疫系統具有很強的抑制作用。近期的研究指出藉由雞卵白蛋白(OVA)引發免疫反應的動物實驗中,TCDD會影響第二型 T 細胞所誘發的細胞激素。因此,本篇研究的主要目的在於探討合併處理TCDD和PCP對於免疫抑制的影響。以雌性BALB/c小鼠進行實驗,先給予OVA腹腔注射,再以灌食的方式給予一次劑量的TCDD或多次劑量的PCP,分別進行七天(給予三次PCP的劑量)及十四天(給予六次PCP的劑量)。實驗結果指出,單獨處理TCDD會降低脾臟對體重的比值,合併處理TCDD和PCP,則會更加減少脾臟對體重的比值,此與脾臟細胞數的計數結果吻合。在七天的組別中,合併處理TCDD及PCP,脾臟中含有表面抗原標記CD4、CD8的細胞數比例降低;在十四天的組別,則含有表面抗原標記CD19的細胞數比例增加。合併處理TCDD和PCP,會抑制OVA刺激的脾臟細胞增生反應。不論單獨給予TCDD、PCP或是合併處理皆會減少由OVA刺激誘發的細胞激素:IL-2、IFN-r、IL-4、IL-5及IL-10的生成,也能抑制免疫球蛋白IgG和IgM的分泌。七天的實驗顯示,單獨處理PCP 與合併處理的組別,IL-4的表現量有上升的趨勢,此現象可能為PCP抑制自然殺手細胞(NK cell)活性的原因。根據上述的實驗結果,PCP能夠影響TCDD的免疫抑制作用,並且在合併的組別中,有許多特別的現象值得進一步去討論並釐清原因。
Pentachlorophenol (PCP) and dioxin have been known to be the famous environmental contaminants. Technical grade PCP contains chlorinated compounds such as chlorodibenzo-p-dioxins is concerned the major cause of suspected immunotoxicity. 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) has been found to be an extremely toxic congener among the halogenated aromatic hydrocarbons and it is a highly immunosuppressive agent. Recent reporters suggest TCDD affects type-2 T cell cytokine productions with ovalbumine (OVA) stimulated immune response in laboratory animals. Hence, the present study is aimed to investigate the combined effect of PCP and TCDD on immunosuppression. Female BALB/c mice were used in this study and they were immunized with OVA and gavaged with TCDD once and PCP in three times a week, separately for 7 days and 14 days. The results indicated that treated with TCDD alone reduced the ratio of spleen to body weight and treated with TCDD in combination with PCP reduced the ratio more than TCDD alone. Splenocyte count showed the same trend with spleen/body weight ratio. In splenocytes subsets, combined treatment decreased the percentage of CD4 and CD8 in splenocytes on day 7, but increased percentage of CD19 in splenocytes on day 14. Combined treatment suppressed OVA-stimulated splenocytes proliferation. TCDD alone, PCP alone and combined treatment suppressed OVA- stimulated cytokines IL-2, IFN-r, IL-4, IL-5 and IL-10 production. In addition, the immunoglobulin, IgG and IgM were suppressed also. On day 7, IL-4 production was increased in mouse treated with PCP alone or in combination; it might result in PCP inhibit the nature killer (NK) cell activity. The current study demonstrated that PCP could affect the immunosuppressive effects of TCDD. However, the detail mechanisms need to be further investigated.
第一章、序論 1
第二章、文獻回顧 2
第一節、 環境污染物與免疫毒性反應 2
第二節、 五氯酚 (Pentachlorophenol) 3
第三節、 五氯酚的毒性與代謝 4
第四節、 戴奧辛與2,3,7,8-四氯戴奧辛(2,3,7,8-Tetrachlorodibenzo-p-dioxin ,TCDD) 5
第五節、 2,3,7,8-四氯戴奧辛的毒性與代謝 7
第六節、 免疫系統(Immune system) 9
第三章、研究目的 16
第四章、研究架構 17
第五章、研究材料與方法 18
第一節、研究材料 18
1. 藥品試劑 18
2. 常用儀器 19
3. 常用溶液 20
第二節、研究方法 21
一、 體內實驗(in vivo) 21
二、 OVA配置 22
三、 Trypan blue dye exclusion assay (計數細胞存活率) 22
四、 分離脾臟細胞 23
五、 脾臟細胞表面標記分析 24
六、 脾臟細胞增生能力分析 25
七、 脾臟自然殺手細胞活性分析 26
八、 細胞激素分析 28
九、 免疫球蛋白表現分析 30
第三節、統計分析 33
第六章、實驗結果 34
第一節、 實驗過程中雌性BALB/c 小鼠的體重與器官重量變化情形
(一) 體重變化 34
(二) 器官重量與體重的比值 34
第二節、 BALB/c 小鼠脾臟細胞數與脾臟內免疫細胞族群百分比的變化
(一) 脾臟細胞數 35
(二) 脾臟細胞表面抗原分析 35
第三節、 BALB/c 小鼠脾臟內免疫細胞功能的變化
(一) 脾臟細胞增生能力(Proliferation) 36
(二) 細胞激素的分泌:(IFN-r,IL-2,IL-4,IL-5,IL-10) 36
(三) 自然殺手細胞活性 37
(四) 血清中免疫球蛋白IgG與IgM的含量 38
第七章、討論 39
第八章、結論及建議 44
第九章、參考文獻 45

圖表 55
附錄 70
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