大鼠小肝细胞移植对急性肝损伤的影响

目的 探讨小肝细胞(small heptocytes,SHCs)移植对四氯化碳(carbon tetrachloride,CCl4)+2/3肝切(partial hepatectomy,PH)所致急性肝损伤的影响.方法 选择健康SD大鼠20只,饲喂含二乙基亚硝胺(DEN)饲料建立SD大鼠SHCs增殖模型.分离、培养、鉴定(电镜、免疫细胞化学)原代SHCs.SHCs行PKH26膜荧光染色.将实验用同一近交系SD大鼠30只随机平均分成空白对照组(B)、损伤造模组(M)和SHCs移植组(T).B组腹腔注射生理盐水.M组、T组腹腔注射CCk.24 h后戊巴比妥腹腔麻醉,消毒开腹.M、T组行2/3肝切,B、M组门静脉注入生理盐水,T组门静脉注入经膜荧光标记SHCs悬液.4周后.采血测血清谷丙转氨酶(ALT)、谷草转氨酶(AST),并取病理标本做冰冻切片荧光显微镜下观察及HE染色普通光镜下观察.结果 电镜下观察可见SHCs表面少量短而小的微绒毛突起,核浆比例高.呈现未分化细胞的形态.免疫细胞化学观察SHCs胞质AFP、CK-19染色呈棕黄色阳性信号.SHCs移植可以显著降低CCl4+2/3PH致肝损伤血清ALT、AST水平,可以明显改善CCl4+2/3PH致肝损伤肝脏病理改变.T组冰冻切片荧光显微镜下可以观察到由SHCs分化的肝细胞形成的肝板结构.结论 SHCs移植对CCl4+2/3PH所致急性肝损伤有一定治疗作用.

Effect of rat small hepatacyte transplantation on acute liver injury

Objective To study the effect of SHCs transplantation on liver injury induced by carbon tetrachloride (CCl4) and 2/3partial hepatectomy (PH). Methods Twenty healthy SD rats were selected to be fed with the bait vessel which contained diethylinitrosamine (DEN) to establish the model of SHCs prolif-eration. After the SHCs were isolated, cultivated and identified, they were stained with PKH26 membrane fluorchrome. The same line 30 SD rats were randomly divided into the blank control group (B), CCl4+ 2/3PH injured group(M) and SHCs transplantation group (T). Rats of group B were intraperitoneally injected with normal saline and those in group M and group T were intraperitoneally injected with CCl4. Twenty-four hours later, all rats were anaesthetized by intraperitoneal injection of butaylone. Then disinfection and opera-tion were done. 2/3PH was done in group M and T. Then the rats in B group and M group were injected normal saline via portal vein, and those in T group were injected with SHCs suspension. Four week later, the level of serous alanine aminotransferase(ALT)and glutamic oxalacetic transaminase (AST) were tested, and liver pathological changes observed. Frozen sections of T group were observed under fluorescent micro-scope. Result Under electron microscope, the manipulus short microvilli on the SHCs' surface and high karyoplasmic ratio were observed as the presentation of dedifferentiation. Immunocytochemistry of alpha fe-toprotein (AFP) and cytokeratin19 (CK-19) staining presented the color of buffy as positive. SHC trans-plantation lowered the levels of serous ALT and AST and significantly improved the degree of injury of path-ologieal section. Under fluorescence microscope, hepatic cell which came from the SHCs constructing hepatic plate were observed. Conclusion Transplantation of SHCs has therapeutic effect on acute liver injury in-duced by CCl4+2/3PH.