- 期刊
黃麴毒素B1對大鼠腹腔沖洗液細胞之致害：DNA單股斷裂及細胞凋亡之誘發
Cytotoxic Effects of Aflatoxin B1 on Rat Peritoneal Lavage Cells: Induction of DNA Single Strand Breaks and Apoptosis
摘要
本研究探討黃麴毒素B1(AFB1)在多重濃度及時間之共同培養條件下，對大鼠腹腔沖洗細胞(RPLC)之致害機制。實驗結果顯示，AFB1(1、50、100µg/mL)與細胞共同培養1-48小時後之毒殺效果呈現顯著時間及濃度效應；應用單細胞電泳試驗檢查AFB1致害細胞DNA單股斷裂程度，也出現顯著之時間效應。RPLC與AFB1共同培養18小時後以電泳法分析細胞DNA，則呈現出185-200 bps DNA片段之規則斷裂形式，證實AFB1可造成細胞凋亡(apoptosis)之現象。在光學及穿透式電子顯微鏡檢視下，50 µg/mL AFB1與RPLC培養，12小時後即可出現RPLC壞死及apoptosis的特徵性變化。然而在50 µg/mL AFB1共同培養0-24小時後之RPLC，並無法檢測到p53及Bcl-2蛋白之表現。本研究結果顯示，AFB1參與並導致DNA斷裂，並使細胞進入細胞凋亡之途徑。而p53及Bcl-2蛋白之表現並不受AFB1之影響；其參與AFB1引發RPLC apoptosis過程之機制則有待進一步探討。
並列摘要
DNA single strand breaks and apoptosis in Aflatoxin B1 (AFB1)-induced cytotoxicities in rat peritoneal lavage cells (RPLC) were investigated. The results of trypan blue dye exclusion assay indicated that significant cytotoxic effects were obvious when RPLC were exposed to 10, 50 and 100 µg/mL AFB1 for 1 and up to 48 hours, and the effects were in dosage and time-dependent manners. These cells also exhibited significant increases in DNA single strand breaks by SCGE assay in dosage-dependent patterns. These AFB1 treatments were also capable of inducing 185-200 bp DNA ladder fragmentation after 18 hours co-incubation. Light and electron microscopic analysis confirmed that RPLC for both 12 and 24 hours could process to necrosis and apoptosis on the same RPLC sample by AFB1 treatment at 50 µg/mL. Western blot analysis of total cellular protein revealed that the RPLC did not express p53 and Bcl-2 proteins after exposed to 50 µg/mL AFB1 for 0, 6, 12 and 24 hours. In conclusion, cellular toxicities including apoptosis and necrosis in RPLC could be the consequences of nucleic DNA damages which were resulting from AFB1 exposure. The role of Bcl-2 and p53 in AFB1 toxicities remained to be determined in the future studies.
延伸閱讀
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