大鼠肝卵圆细胞增殖模型的建立与优化

目的 建立大鼠肝卵圆细胞增殖模型，并观察2-乙酰氨基芴（2-acetaminofluorene,AAF)剂量与模型动物肝卵圆细胞增殖程度间的关系。方法 选择体重150g左右的雄性Wistar大鼠，通过胃管灌喂AAF，每日1次，连续4d，第5日行三分之二肝切除（手术当日不灌喂AAF），第6日继续灌喂，连续1周。AAF剂量分别按2.5、5、10、20mg/kg给予，对照组给予生理盐水灌喂。各组手术后复隔2-3d分别取3只大鼠肝组织作常规组织学观察及免疫组织化学染色。结果 对照组大鼠肝组织内未见到肝卵圆细胞，2.5mg/kg剂量组及5mg/kg剂量组仅见少量肝卵圆细胞增生，而10、15、20mg/kg三个剂量组肝组织内均见明显的肝卵圆细胞增殖反应；肝卵圆细胞胞浆细胞角蛋白19（CK19）、OV6及波形蛋白染色均呈阳性，胞核增殖细胞抗源染色呈阳性。结论 AAF剂量为10-20mg/kg时可获得满意的大鼠肝卵圆细胞增殖模型。

Establishment and optimization of rat models for hepatic oval cells proliferation

ve To establish a rat model for hepatic oval cell proliferation and to observe the relationshipbetween 2-acetaminofluorene (AAF) dosage and oval cell proliferation in the rat liver. Methods Male Wistar rats weighingl50 g received daily oral gavage of AAF for 4 days before operation and up to 7 days after operation. Two-thirds hepatec-tomy was performed on the 5th day and the gavage was not performed on the day of operation. AFF was given with thedosage of 2.5 mg/kg, 5 mg/kg, l0 mg/kg, l5 mg/kg, and 20 mg/kg body weight.Animals in control group were given saline.Three rats from each group were killed every 2~3 days after hepatectomy and liver slices were fixed and processed for rou-tine histology and immunohistochemistry. Results Hepatic oval cells were not observed in the liver of controls and only afew were detected in the liver of 2.5 mg/kg and 5 mg/kg groups. However, obvious oval cell proliferation was seen in theliver of l0 mg/kg, l5 mg/kg, and 20 mg/kg groups. Hepatic oval cells were stained positive for cytokeratin l9, OV6,vimentin and proliferating cell nuclear antigen (PCNA). Conclusions Satisfactory rat models for hepatic oval cell prolif-eration can be obtained using our scheme when AAF is dosed at 10~20 mg/kg body weight.