State Key Laboratory for Diagnosis and Treatment of Infectious Disease, First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China.
Hepatobiliary Pancreat Dis Int. 2010 Dec;9(6):611-4.
The bioartificial liver is anticipated to be a promising alternative choice for patients with liver failure. Toxic substances which accumulate in the patients' plasma exert deleterious effects on hepatocytes in the bioreactor, and potentially reduce the efficacy of bioartificial liver devices. This study was designed to investigate the effects of plasma from patients with acute on chronic liver failure (AoCLF) on immortalized human hepatocytes in terms of cytochrome P450 gene expression, drug metabolism activity and detoxification capability.
Immortalized human hepatocytes (HepLi-2 cells) were cultured in medium containing fetal calf serum or human plasma from three patients with AoCLF. The cytochrome P450 (CYP3A5, CYP2E1, CYP3A4) expression, drug metabolism activity and detoxification capability of HepLi-2 cells were assessed by RT-PCR, lidocaine clearance and ammonia elimination assay.
After incubation in medium containing AoCLF plasma for 24 hours, the cytochrome P450 mRNA expression of HepLi-2 cells was not significantly decreased compared with control culture. Ammonia elimination and lidocaine clearance assay showed that the ability of ammonia removal and drug metabolism remained stable.
Immortalized human hepatocytes can be exposed to AoCLF plasma for at least 24 hours with no significant reduction in the function of cytochrome P450. HepLi-2 cells appear to be effective in metabolism and detoxification and can be potentially used in the development of bioartificial liver.
生物人工肝被期望成为肝功能衰竭患者的一种有前途的替代选择。积聚在患者血浆中的毒性物质对生物反应器中的肝细胞产生有害影响，并可能降低生物人工肝设备的疗效。本研究旨在研究急性肝衰竭（AoCLF）患者血浆对永生化人肝细胞的细胞色素 P450 基因表达、药物代谢活性和解毒能力的影响。
永生化人肝细胞（HepLi-2 细胞）在含有胎牛血清或来自 3 名 AoCLF 患者的人血浆的培养基中培养。通过 RT-PCR、利多卡因清除率和氨消除测定评估 HepLi-2 细胞的细胞色素 P450（CYP3A5、CYP2E1、CYP3A4）表达、药物代谢活性和解毒能力。
在含有 AoCLF 血浆的培养基中孵育 24 小时后，HepLi-2 细胞的细胞色素 P450 mRNA 表达与对照培养相比没有明显降低。氨消除和利多卡因清除试验表明，氨去除和药物代谢的能力保持稳定。
永生化人肝细胞可以暴露于 AoCLF 血浆中至少 24 小时，而细胞色素 P450 的功能没有明显降低。HepLi-2 细胞在代谢和解毒方面似乎有效，并且可潜在用于生物人工肝的开发。
