An in vivo method for using 5-bromo-2′-deoxyuridine (BrdU) as a marker of chemically-induced hepatocellular proliferation in the Japanese medaka (Oryzias latipes)
An in vivo method for using 5-bromo-2′-deoxyuridine (BrdU) as a marker of chemically-induced hepatocellular proliferation in the Japanese medaka (Oryzias latipes)
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DOI：
10.1080/019262301316905354
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发表时间：
2001-05-01
影响因子：
1.5
通讯作者：
Gardner, HS
中科院分区：
文献类型：
--
作者：
Brennan, LM;Boncavage-Hennessey, EM;Gardner, HS
以日本青花鱼(Oryzias Latipe)为材料，建立了一种体内方法来评估非哺乳类动物模型中的肝细胞增殖。分别于染毒后7天、17天、24天、94天观察青竹细胞在10 mg/L二乙基亚硝胺(DEN)作用48小时后的增殖反应。青山鼠暴露于50或75 mg/L的5-溴-2‘-脱氧尿嘧啶核苷(BrdU)水中72小时后，处死动物，进行免疫组织化学染色。BrdU标记的肝细胞的增殖指数通过计数和面积测量进行量化和比较。在前3个时间点，100 mg/L DEN处理组的肝细胞增殖显著高于对照组和10 mg/L处理组。在DEN暴露26周后评估其肝癌致癌性。与其他鱼相比，100 mg/L DEN处理组的肝细胞肿瘤发生率显著增加(p<；0.0001)。在水族箱水中加入BrdU，可有效地标记青蛙S期肝细胞，并在暴露于致癌浓度的DEN后7天检测到肝细胞增殖的增加。此外，新的面积测量方法的增殖指数与计数指数(R(2)大于或等于0.92)一样精确。
Japanese medaka fish (Oryzias latipes) were used to develop an in vivo method to assess hepatocellular proliferation in a nonmammalian model. Proliferative responses were assessed in medaka at 7, 17, 24, and 94 days after a 48-hour exposure to 10 or 100 mg/L diethylnitrosamine (DEN). Subgroups of medaka were exposed to 50 or 75 mg/L of 5-bromo-2'-deoxyuridine (BrdU) in water for 72 hours, sacrificed, and then processed for immunohistochemical staining. Proliferative indices of BrdU-labeled hepatocytes were quantified and compared using both count and area measurements. There was a significant increase (p < 0.05) in hepatocellular proliferation in the 100 mg/L DEN-treated fish as compared to controls and 10 mg/L DEN-treated fish for the first 3 time points. Hepatocarcinogenicity was evaluated 26 weeks post-DEN exposure. There was a significant increase (p < 0.0001) in hepatocellular neoplasms in 100 mg/L DEN-treated fish compared to other fish. Effective BrdU-labeling of S-phase hepatocytes in medaka was achieved by adding BrdU to the aquarium water, and an increase in hepatocellular proliferation using this method was detected 7 days after exposure to a carcinogenic concentration of DEN. Additionally, the new method of area measurement indices of proliferation were as precise as count indices (R(2)greater than or equal to0.92).