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題 名 | 黃麴毒素對鴨淋巴細胞轉型作用之影響 |
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作 者 | 鄭永祥; 龐飛; | 書刊名 | 中華農學會報 |
卷 期 | 170 1995.06[民84.06] |
頁 次 | 頁147-155 |
分類號 | 437.22 |
關鍵詞 | 黃麴毒素; 淋巴細胞轉型作用; 鴨; Aflatoxins; Lymphocyte transformation; Duck; |
語 文 | 中文(Chinese) |
中文摘要 | 本研究之目的在於嘗試自周邊血液分離及純化鴨淋巴細胞,探測其對各種淋巴細胞裂殖原刺激之反應,並藉以探討黃麴毒素對鴨淋巴細胞轉型作用之影響。 將EDTA抗凝之成熟產蛋菜鴨血,於4℃下,1300×g,離心30分鐘,取Buffy coat,經等量PBS稀釋後,置於Ficoll-Paque上層,於4℃下,200×g,離心25分鐘,取界面層可得鴨淋巴細胞再經PBS沖洗一次,RPMI清洗培養液沖洗兩次,每次均於4℃,200g,離心10分鐘。細胞經cytospin後製成抹片,以Quick stain染色計數,所得鴨淋巴細胞純度可達95%以上,平均每10ml血液約可分離2×10⁷細胞。取鴨淋巴細胞(8×10⁵ /mL)分別加入最終濃度為0、5、10、20、40或80 μg/ml之裂殖原(mitogen),包括有PWM、PHA、PNA及BSS;經培養72小時後,以³H-thymidine (1 μCi/well)併入法測定鴨淋巴細胞轉型(lymphocyte transformation)反應。結果以PHA 5及10 μg/ml濃度下所測得CPM值高於其他裂殖原。 為探討黃麴毒素對鴨淋巴細胞轉型作用之影響,取鴨淋巴細胞(8×10⁵ cells/ml),以PHA (100 μg/ml)為陽性對照組,分別加入10⁴、10¹或10⁻² ng/ml的黃麴毒素B₁;為瞭解微粒體代謝作用對黃麴毒性之影響,於各測試組中加入有或無鴨肝臟微粒體(1 mg/ml)與NADPH (0.01mM)二組,經培養後,結果以黃麴毒素10¹及10⁻² ng/ml測得之CPM值顯著高於其他各組(P<0.05),顯示低濃度黃麴毒素對鴨淋巴細胞增殖具有刺激作用,輕微的抑制現象僅在高濃度10⁴ ng/ml下始出現。反之,當有微粒體存在時,即使在低濃度10⁻² ng/m下對鴨淋巴細胞轉型作用即有負面影響,而此抑制作用隨黃麴毒素濃度升高而愈明顯。因此,黃麴毒素在經由肝臟微粒體的代謝後,可顯著的抑制PHA所引發的鴨淋巴細胞轉型作用。 |
英文摘要 | The purpose of this experiment was to isolate and purify duck lymphocytes from peripherial blood. Lymphocytes were then used to study transformation response to various mitogens. These results were used to evaluate the effects of aflatoxin on duck lymphocyte transformation. The buffy coat collected from the whole blood was anticoagulated with EDTA andcentrifuged at 1300×g for 30min at 4°C, then resuspended with an equal wolume of PBS. The cell suspension was layered over a Ficoll-Paque gradient and centrifuged at 200×g for 25min at 4°C. The band formed at the interface of the RBC layer contained cells of which greater than 95% were lymphocytes. The averaged numbers of lymphocytes obtained from 10 mL of whole blood wasapproximately 2×10⁷. various mitogens (PWM, PHA, PNA, and BSS) at a final concentration of 9, 5, 10, 20, 40 and 80μg/mL were added to the lymphocyte suspension (8×10⁵ cells/mL). The CPM value of the lymphocyte transformation response by ³H-thymidine integration after 72hrs incubation was measured. The results showed that PHA at 5 and 10μg/mL gave higher values than the others. The effects of aflatoxin on duck lymphocyte transformation response were evaluated, by addition of final concentration of 10⁴, 10¹, or 10⁻²ng/mL aflatoxinB₁ (AFB₁) to lymphocyte suspension. Meanwhile, lymphocyte suspension with or without duck liver micorosome plus NADPH was also evaluated. The results showed AFB₁ 10¹ and 10⁻²ng/mL had significantly higher CPM values than the others (P<0.05). This indicated that AFB₁ had a stimulative effect on duck lymphocyte transformation. AFB₁ only at 1 10⁴ng/mL had a slight inhibition. When liver microsome was present, AFB₁ had an adverse effect on duck lymphocyte transformation even though at a low concentraction of 10⁻²ng/mL. This detrimental effect became more-obvious as AFB₁ concentration increases. It could thus be concluded that PHA-induced duck lymphocyte transformation was inhibited significantly when was metabolized by duck liver micorsome. |
本系統中英文摘要資訊取自各篇刊載內容。