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我们通过制备实验性亚急性肝功能衰竭模型,运用免疫组化技术,检测再生肝细胞表面增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)表达分布状态,旨在对肝细胞再生程度作一动态评价。 1 材料和方法 1.1 材料 亚急性肝功衰竭模型的制备,参照Ohnishi et al方法并加以改进,Wistar (?)大白鼠(中国医科大学第二临床学院实验动物室提供)110只,体质量200g,先取50只,分5组,每组10只,选用内毒素同型物FS-112(购自Ono Phamaceutical Co Japan),以30mg/kg(溶于50mL/L乙二醇,50mL/L乙醇,50g/L葡萄糖)经尾静脉注入,2d后行70％部分肝切除术,分别在术后12,24,36,48,72h进行心脏采血,3000r/min离心10min,分离血清,-20℃保存待检,之后立即开腹留取剩余肝
By preparing experimental sub-acute liver failure model and using immunohistochemical technique to detect the expression and distribution of proliferating cell nuclear antigen (PCNA) on the surface of regenerating hepatocytes, we aimed to make a dynamic evaluation of the degree of hepatocyte regeneration. 1 Materials and methods 1.1 Materials Sub-acute hepatic failure model was prepared according to Ohnishi et al method and improved, Wistar (?) Rats (Experimental Animal Room, Second Clinical College, China Medical University) 110, body weight 200g, Fifty rabbits were randomly divided into five groups with 10 mice in each group. Endotoxin homogenate FS-112 (purchased from Ono Pharmaceutical Co., Japan) was used at a concentration of 30 mg / kg in 50 mL / L ethylene glycol, 50 mL / / L glucose) was injected through the tail vein, 2 days after 70% partial hepatectomy, respectively, after 12,24,36,48,72 h cardiac blood sampling, 3000r / min centrifugation 10min, serum was isolated and stored at -20 ℃ Check immediately after opening the remaining liver