中国血液净化

›› 2006, Vol. 5 ›› Issue (9): 677-679.

• 基础研究 • 上一篇    下一篇

冻存时间不同的成年动物肝细胞复苏后对重型肝炎胆红素代谢的效果比较

黄庆山 陈南清 伍通和 李 峥 石添元 詹金美   

  1. 364000 龙岩,福建省龙岩市第二医院人工肝治疗中心
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2006-09-12 发布日期:2006-09-12

  • Received:1900-01-01 Revised:1900-01-01 Online:2006-09-12 Published:2006-09-12

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摘要： 目的 实验比较在液氮中冻存时间长短不同的成年动物肝细胞(adult animal hepatocytes,AAH)复苏后对人体重型肝炎胆红素代谢的效果，探讨体外生物人工肝支持系统(extracorporeal bioartificial liver support system，EBLSS)使用冻存复苏AAH的实际意义。方法 采用HE染色镜检肝细胞形态、台盼兰染色计数肝细胞成活率、肝细胞培养并贝克曼生化分析仪检测对重型肝炎胆红素的代谢效果。实验观察液氮中冻存1周、1个月、3个月复苏后的成年小白鼠肝细胞形态、存活率和对重型肝炎胆红素的代谢效果，同时设新鲜分离的肝细胞作对照组。结果 冻存1周、1个月的肝细胞在光镜下形态无明显改变，冻存3个月的肝细胞胞质严重丢失；对照组、冻存1周、1个月、3个月的肝细胞成活率分别为91%、83%、76%、35%(与对照组比较，后三者分别P＞0.05，P＜0.01，P＜0.01)；四组肝细胞使重型肝炎总胆红素分别下降27.01%、23.02%、19.58%、2.33%，(与对照比较，后三者分别P＜0.01，P＜0.001，P＜0.001)。结论 EBLSS以使用新鲜分离的AAH为佳。

关键词: EBLSS, 肝细胞, 冻存复苏, 存活率, 胆红素

Abstract: Objective To compare the effect of adult animal hepatocytes after resuscitation with different cryopreservation time on bilirubin metabolism in severe hepatitis for investigating its practical significance on the extracorporeal bioartificial liver support system (EBLSS). Methods The morphologic changes of the adult mice hepatocytes were observed and compared after resuscitation with liquid nitrogen cryopreservation for one week, one month, three months under microscopy by HE staining, count the survival rate of hepatocytes were calcatated according to trypan blue staining and the metabolic activity of bilirubin of severe hepatitis were determined with cell culture and Beckmann auto-analyzer, the fresh-separated hepatocytes were regarded as control group. Results The cytoplasm of hepatocytes after cryopreservation resuscitation for three months lost markedly, and no change for one week and one month. The survival rates of hepatocytes in control group, cryopreservation for one week, one month, three months were 91%, 83%, 76% and 35% respectively, the total bilirubins of severe hepatitis of four groups fell 27.01%, 23.02%, 19.58% and 2.33% respectively. Conclusion The fresh-separated adult animal hepatocytes is good in EBLSS.

Key words: Hepatocyte, Cryopreservation Resuscitation, Survival rate, Bilirubin

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