Wu Chen-Xuan, Zou Qi, Zhu Zheng-Yan, Gao Ying-Tang, Wang Yi-Jun
Department of Hepatobiliary, The Third Center of Affiliated Hospital of Tianjin Medical University, Key Laboratory of Artificial Cells of Tianjin, Tianjin 300170, China.
World J Gastroenterol. 2009 Mar 28;15(12):1506-11. doi: 10.3748/wjg.15.1506.
To evaluate the effect of intrahepatic trans-plantation of hepatic oval cells (HOC) on fulminant hepatic failure (FHF) in rats.
HOC obtained from rats were labeled with green fluocescent protein (GFP) or 5, 6-carboxyfluorescein diacetate succinmidyl ester (CFDA-SE). Cell fluorescence was observed under fluorescent microscope at 6, 24, 48 and 72 h after labeling. CFDA-SE labeled HOC (5 x 10(6) cells each rat) were injected into livers of rats with FHF induced by D-galactosamine. Serum albumin (ALB), alanine aminotransferase (ALT), aspartate aminotransferase (AST) and total bilirubin (TBil) levels were measured at different time points. Liver function of rats was examined on days 3, 7, 14 and 21 after HOC transplantation.
The positive rate of GFP and CFDA-SE labeled HOC was 10% and 90%, respectively, with no significant change in cell viabilities. The survival rate was higher in HOC transplantation group than in control group, especially 48 (9/15 vs 6/15) and 72 h (9/15 vs 4/15) after HOC transplantation. The serum ALT, AST and TBil levels were decreased while the serum Alb level was increased after HOC transplantation. Fluorescence became faded and diffused in liver tissues, suggesting that proliferation and differentiation occur in transplanted HOC.
CFDA-SE is superior to GFP in labeling HOC, although fluorescence intensity is decreased progressively with cell division. HOC transplantation can improve the liver function and increase the survival rate of recipients.
评估肝卵圆细胞(HOC)肝内移植对大鼠暴发性肝衰竭(FHF)的影响。
从大鼠获取的HOC用绿色荧光蛋白(GFP)或5,6-羧基荧光素二乙酸琥珀酰亚胺酯(CFDA-SE)标记。标记后6、24、48和72小时在荧光显微镜下观察细胞荧光。将CFDA-SE标记的HOC(每只大鼠5×10⁶个细胞)注入用D-半乳糖胺诱导FHF的大鼠肝脏。在不同时间点测量血清白蛋白(ALB)、丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)和总胆红素(TBil)水平。在HOC移植后第3、7、14和21天检查大鼠肝功能。
GFP和CFDA-SE标记的HOC阳性率分别为10%和90%,细胞活力无显著变化。HOC移植组的存活率高于对照组,尤其是在HOC移植后48小时(9/15比6/15)和72小时(9/15比4/15)。HOC移植后血清ALT、AST和TBil水平降低,而血清Alb水平升高。荧光在肝组织中变淡并扩散,表明移植的HOC发生了增殖和分化。
CFDA-SE在标记HOC方面优于GFP,尽管荧光强度随细胞分裂逐渐降低。HOC移植可改善肝功能并提高受体的存活率。