摘要
目的设计一种新型混合型生物人工肝(HBAL)并通过对模拟肝衰竭血清的净化作用评价其疗效,探讨其临床应用的可行性。方法选用中国人肝细胞株-1(CL-1)作为肝细胞供体,在微重力环境下,肝细胞微载体共培养方法培养至第5天,细胞总量约4.0×109个,细胞密度约为4.0×1010/L,在无菌环境下灌入自制生物反应器中,制成生物部分,非生物部分采用血液灌流+胆红素吸附,生物部分和非生物部分通过两套聚乙烯胶管构成一个封闭的环路,监测模拟肝衰竭血清中未结合胆红素、鹅去氧胆酸,胆酸、血氨经过HBAL循环后浓度的变化以及生物反应器中肝细胞功能、形态及肝细胞活性的变化。结果整个治疗过程中循环前24h内,模拟肝衰竭血清中未结合胆红素(UBD)、鹅去氧胆酸(CDCD),胆酸(CA)和血氨(AA)的浓度分别从(335.3±6.0)、(395.0±5.6)、(155.7±4.5)、(39.0±2.6)μmol/L下降至(106.0±10.9)、(131.8±28.7)、(42.2±7.3)、(3.5±1.0)μmol/L,与0h比较差异有统计学意义(P=0.041),24h后浓度下降趋于平稳;循环48h后,CL-1细胞功能发生变化,生物反应器中模拟肝衰竭血清中的谷丙转氨酶(ALT)、谷草转氨酶(AST)、乳酸脱氢酶(LDH)分别从(25.9±4.2)IU/L、(22.0±3.6)IU/L、(0.28±0.09)μmol/L升高至(31.0±2.6)IU/L、(31.6±8.0)IU/L、(0.41±0.12)μmol/L,与0h比较差异有统计学意义(P=0.027);并且整个反应器中肝细胞的数量和活力在循环48h后也呈现显著性下降(P=0.044)。结论(1)构建了一种新型灌流型生物反应器,该生物反应器中的CL-1细胞能够保持较高的活性和良好的功能。(2)新型混合生物型人肝可以显著降低模拟肝衰竭血清中的未结合胆红素、鹅去氧胆酸,胆酸、血氨的浓度,具有清除这些毒性物质的功能,提示其具有明显的肝功能支持作用。
Objective To design a new type of hybrid bioartificial liver and evaluated its efficacy in vitro,explore the feasibility of its clinical application.Methods CL-1 human hepatocytes were cultured on microcarries to fiith day,when cells is about 4.0 × 109 and cell density of about 4.0 × 1010/L,CL-1 cells cultured on microcarriers and self-made bioreactor constitute biological part of the hybird biological artificial liver,Abiotic part includes blood perfusion and bilirubin adsorption,blood pump employed as circulation driver,which were composed of a hybrid bioartificial liver,the changes of concentrations of indirect bilintbin,chenodeoxycholic acid,bile acid,blood ammonia,aspartate aminotransferase (AST),alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) were observed under the condition of in vitro circulation.Meanwhile,CL-1 cell function,morphology and the cell activity were also observed.Results After 24 h of the in vitro circulatioan,the concentration of indirect bilirubin (UBD) and chenodeoxycholic acid (CDCD),cholic acid (CA) and of ammonia (AA) respectively decreased from (335.3±6.0),(395.0±5.6),(155.7±4.5),(39.0±2.6) μmol/L to (106.0±10.9),(131.8± 28.7),(42.2 ± 7.3),(3.5 ± 1.0) μmol/L,have statistical significance compared with 0 (P =0.041),48 h later the degree of ALT,AST and LDH significantly increased from (25.9 ±4.2) IU/L,(22.0 ± 3.6) IU/L,(0.28 ± 0.09) μmol/L to (31.0 ± 2.6) IU/L,(31.6 ± 8.0) IU/L,(0.41 ± 0.12) μmoL/L,meanwhile the number of CL-1 cells and its vitality were significant decline (P =0.027,0.044).Conclusion We therefore conclude that (1) In the newly hybrid bioartificial liver (HBAL) system,CL-1 cells can keep its viability and function in vitro,and (2) the HBAL appears to be effective in purified the simulation serum of liver failure,with improvement non-conjugated bilirubin,chenodeoxycholic acid,cholic acid and ammonium chloride,this HBAL system seems to be promising.
作者
尹清华
邵梅
高毅
高森
郭颖
Yin QingHua;Shao Mei;Gao Yi;Gao Sen;Guo ling(Department of General Surgery,the First Hospital of Changsha,Changsha 410000,China;Department of General Surgery,Penglai Hospital Affiliated to Taishan Medical University,Penglai 264000, China;Cardio-Thoracic Surgery,Zhujiang Hospital of Southern Medical University,Guangzhou 510000,China;Department of General Surgery,the Second People's Hospital of Hefei,Hefei 230000,China)
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2018年第12期2256-2260,共5页
Chinese Journal of Experimental Surgery
基金
国家高技术发展计划(863计划)(2006AA02A141).
