高原体外生物型人工肝装置的构建与初步评价

Establishment and preliminary evaluation of bioartificial liver device at high altitude region

目的在高原环境条件下构建体外生物型人工肝装置,通过在线运行初步评价其安全性和稳定性,为应用于治疗急性肝衰竭实验研究提供依据。方法采用已适应本地环境的健康长白仔猪2头,雌雄不限,体质量分别为29.5 kg和31.0 kg。建立经颈外静脉至下腔静脉/颈外静脉插管组合血路,以驱动泵、血流管路、辅助加热器及含有培养肝细胞的生物反应器构建生物型人工肝装置,于体外血液循环中持续转流8 h,观察实验前后动物的一般状态、心率、血常规、尿常规及血液生物化学指标改变,有无出血倾向发生,血流管路凝血情况及培养肝细胞活性的变化。结果分离所得肝细胞量为2.72×109,肝细胞纯度为93%,肝细胞活率为96%。体外血液循环实验后,动物未出现明显不良反应,心率无显著改变,血常规、尿常规及血液生物化学指标均正常,未见出血倾向;血流管路凝血不明显;反应器内培养肝细胞活率,实验前后分别为(87.33±2.08)%和(84.67±3.79)%,差异无统计学意义(P>0.05)。结论该方法构建的高原体外生物型人工肝装置较为稳定、可靠,动物耐受性好。

Objective To establish a bioartificial liver device（BALD） and evaluate its safety and stability at high altitude region,and provide the basis for experimental research on the treatment of acute liver failure.Methods Two local healthy pigs were enrolled,with body weight of 29.5 kg and 31.0 kg.The combination trail via external vein to inferior vena cava/external vein were constructed,and BALD was composed of driving pumps,blood flow line,auxiliary heater and biological reactor containing pig hepatocytes.The extracorporeal circulation was performed at a blood current velocity of 400 mL/min for 8 hours,and the general condition,heart rate（HR） and bleeding tendency of the experimental pig,coagulation in the cannula and activity of the hepatocytes in biological reactor were observed.Results The number of the isolated liver cells was 2.72 × 109,with a purity of 93 % and viability of 96 %.No obvious adverse reaction,significant HR change,and bleeding tendency were observed in the experimental animals.Blood coagulation in the cannula was not significant.The viability of the hepatocytes after experiment was not significantly reduced as compared with that before the experiment,which was（84.67 ± 3.79） % and（87.33 ± 2.08） %,respectively（P 0.05）.Conclusion It is demonstrated that the BALD has good stability and safety for pig at high altitude.