闭合型双循环生物人工肝支持系统治疗犬急性肝功能衰竭实验研究

Experimental study on closing bicirculating bioartificial liver support system for treatment of acute hepatic failure in canines

目的探讨和评价闭合型双循环生物人工肝支持系统（CBC-BALSS）在治疗犬急性肝功能衰竭模型过程中的稳定性、安全性和有效性。方法建立犬急性肝功能衰竭模型（门腔分流联合胆总管离断），采用CBC-BALSS进行支持治疗。20只模型犬分为两组：CBC-BALSS治疗组（n=11）；无肝细胞CBC-BALSS对照组（n=9）。治疗时限6h。检测实验犬血氨、生化全套、凝血因子（Factor Ⅶ）、支／芳氨基酸（BCAA／AAA）、单乙基甘氨酸二甲苯胺（monoethylglycinexylidide，MEGX）和细胞循环路生化全套、肝细胞密度和数量。结果CBC—BALSS细胞回路细胞悬液总体积200ml，肝细胞的总数1×10”个、密度5×10^7／ml、活率98％左右。治疗中16只犬的生命体征平稳，在治疗30min内均出现一过性低血压；2只转流开始15min出现过敏反应；1只转流中因上消化道出血死亡；1只因穿刺部位出血死亡。模型治疗前血氨、ALT、TBil／DBil、白蛋白、Factor Ⅶ和BCAA／AAA分别达150mmol／L、400U／L、80／55mmol／L、35g／L、20％和1．6；CBC-BALSS治疗6h后，血氨、TBil／DBil下降均显著低于对照组；ALT存在下降趋势且在第6小时差异有统计学意义；白蛋白、Factor Ⅶ和BCAA／AAA在所有时段、组间差异均无统计学意义。在治疗1h和2h，MEGX差异有统计学意义，治疗组MEGX比对照组提前2h达最高点。治疗15～30min后，双循环路压力至115mm Hg趋于平稳，且在±5mm Hg波动。在治疗过程中，治疗组细胞循环路ALT显著性升高；组间细胞循环路TBil／DBil变化差异无统计学意义，而两组在各时间点均显著性升高；白蛋白变化无统计学意义。结论CBC-BMLSS治疗犬急性肝功能衰竭过程中，安全、有效、稳定且代谢支持作用明显。

Objective To study and evaluate the stability, safety and efficacy of closing bicirculating bioartificial liver support system （CBC-BALSS） in treatment of acute hepatic failure in canines. Methods Twenty canine models with acute hepatic failure were set up through end-side portocaval shunt with common bile duct transaction and then treated with CBC-BALSS （treatment group, n = 11） or no hepatocyte CBC-BALSS （control group, n =9） for six hours. Serum ammonia, biochemical functions, factor Ⅶ, BCAA/AAA and monoethylglycinexylidide （MEGX） in serum of subjects and density, viability and number of hepatocytes in cell circulation were detected in treatment group. Results The total volume of hepatocyte solution was 200 ml, with number, density and viability of hepatocyte for 1 × 10^10 , 5 × 10^7/ml and 98% respectively. During operation, stable clinical vital signs but transient hypotension within 30 minutes were seen in 16 canines including two with allergic reaction after 15 minutes （one per group）, one death due to upper gastrointestinal hemorrhage and one death because of hemorrhage at puncture spot in CBC-BALSS group. Before treatment, serum ammonia, ALT, total/direct bilirubin, factor Ⅶ and BCAA/AAA were 150 mmol/L, 400 U/L, 80/55 mmol/L, 35 g/L, 20% and 1.6, respectively. After treatment for six hours, serum ammonia and total/direct bilirubin decreased more significantly in treatment group than those in control group. ALT decreased significantly and showed statistical difference at the sixth hour. But there was no statistical difference in factor Ⅶ, BCAA/AAA and ALP between two groups at all time points. There was statistical difference in MEGX at the first and second hours, with climax of treatment group arrived earlier two hours. Bicirculating pressure was elevated to 115 mm Hg gradually, fluctuating at ± 5 mm Hg. ALT increased significantly in treatment group. No statistical difference was found two groups upon Tbil/Dbil that increased at all time points. Changes of ALT had no statistical difference. Conclusion ICBC-BALSS is efficacious, safe and stable for treating acute hepatic failure in canines and exerts marked metabolic function.