表达人肝再生增强因子肝细胞系生物人工肝治疗安全性的动物实验初步研究

目的：评价表达人肝再生增强因子基因的HepG2细胞系的细胞培养上清及细胞裂解物的小鼠急性毒性和近期致瘤性。方法：SPF级昆明种小鼠18只，随机分为空白对照组、细胞培养上清组、细胞裂解物组，每组小鼠各6只，腹腔分别接种空白培养液、细胞培养上清、细胞裂解物0．5ml。连续14天，每天观察记录动物毒性反应，14d后宰杀小鼠，取血测血生化指标。及观察病理改变。结果：各组小鼠均存活。除对照组1例小鼠，细胞培养上清组1例小鼠，细胞裂解物组2例小鼠次日活动稍减少外，均未见异常反应。血液生化检测ALT、AST、AFP、TBIL无明显异常，且各组间无差别。普通光镜下各组动物肝脏病理切片染色均未见明显异常。结论：目的细胞系细胞培养上清、细胞裂解物对实验用昆明小鼠无明确毒副作用及短期致瘤性。可能提供一种安全的可用于生物人工肝新的细胞来源。

Study on Safety of HepG2 Cell Line Expressing Augmenter of Liver Regeneration for Bioartificial Liver in Mice

Objective： To evaluate the acute toxicity and short-term tumorigenicity of mice intraperitoneal injection with the culture supernatants or lysates from HepG2 cell line expressed the augmenter of liver regeneration gene. Methods： 18 SPF grade-Kunming mice were randomly divided into 3 groups including control, cell culture supernatant and cell lysate.6 mice in each group were injected intraperitoneally with culture medium, cell culture supernatant, and cell lysate of 0.5ml, respectively. The behaviors of mice in each group were observed each day for two weeks. After two weeks, the mice were killed to measure blood biochemical index and pathological change. Results： All mice were survived throughout the experiment and no abnormal behaviors observed in the mice except for 4 mice （1 in control group, 1 in culture supernatant group and 2 in lysate group）, whose activity slightly decreased. The levels of ALT, AST, AFP, and TBIL were normal and there was no significant difference among three groups. There was no obvious pathological change of liver section from three groups observed by light microscope. Conclusion： There are no acute toxicity and short-term tumorigenicity in experimental SPF-Kunming mice intraperitoneal injection with the culture supematants or lysates from HepG2 cell line expressed the augmenter of liver regeneration gene. The cell line may provide a potential safe cell source for bioartificial liver.