TECA-I型生物人工肝支持系统治疗急性肝衰竭犬的实验研究

Experimental study of type TECAI bioartificial liver suppert system to treatment of acute liver failure in a canine model

目的 :评价经改进的 TECA I型生物人工肝脏支持系统 (bioartificial liver supportsystem ,BAL SS)治疗醋氨酚诱发急性肝衰竭 (acute liver failure,AL F)犬的有效性和安全性。方法 :采用多次皮下注射醋氨酚的方法建立 AL F模型犬。分离中国实验用小型猪肝细胞并培养于 BAL SS中 ,对 AL F犬进行 6小时的治疗 ,观察治疗前后犬生理、生化和组织学的变化 ,与常规药物治疗组和对照组进行比较。结果 :注射醋氨酚 48小时后 ,可建立 AL F犬模型 ,模型成功率为 6 3.16 %。应用我们改进的酶消化法 ,平均从每只小型猪的肝脏可得到(0 .8～ 3.0 )× 10 1 0 个肝细胞且存活率较高。TECA I型 BAL SS对 AL F犬治疗后 ,血氨、丙氨酸转氨酶、天冬氨酸转氨酶和碱性磷酸酶等生化指标明显下降 ,支链氨基酸 /芳香族氨基酸比值显著上升 ,其肝脏的病理改变得以修复 ,治疗后犬可长期存活 (>30日 )。药物治疗组和对照组的 AL F犬病情持续恶化 ,并于治疗后 14.83～6 0 .0 0小时内死亡。结论 :此型 BAL SS能够迅速、安全、有效地暂时替代药物诱发的 AL F犬的肝脏功能 ,其疗效明显优于常规药物治疗 ,BAL SS可望成为救治 AL F的新方法。

Objective:To assess the efficacy and safety of reformed type TECAI bioartificial liver support system(BALSS) on treatment of acute liver failure(ALF)in a canine model induced by acetaminophen. Methods:The ALF canine model by multiple subcutaneous injections of acetaminophen were replicated. The primary hepatocytes were separated from the swine and cultured within the BALSS. The ALF model was treated with BALSS for six hours, the changes in physiological,biochemical and histological parameters were observed before and after the treatment ,and compared them with that after drugs treatment and control groups. Results: The ALF canine model was developed 48 hours after administrate of acetaminophen, the successful rate of model was 63 16%. Using our modified enzymatic digestion method, the yield of hepatocytes was (0 83 0)×10 10 from per swine with high viability. Type TECAI BALSS treatment resulted in beneficial effects with decrease in plasma NH 3, alanine aminotransferase(ALT), aspartate aminotransferase(AST), alkaline phosphatase(ALP) levels decreased,and the ratio of branchchain amino acid to aromatic amino acid( BCAA/AAA ) increased. The pathological changes of the liver were repaired,and the animals with ALF could live longer than 30 days. The conditions of ALF models after the coventional treatment and control groups deteriorated, and they died within 14 8360 00 hours. Conclusions:This type of BALSS is rapid,safe and efficacious in providing liver support in the ALF canine model.The efficacy of BALSS is superior to the routine drugs treatment,and it would hopefully be a new method in the treatment of ALF.