De Smet K, Brüning T, Blaszkewicz M, Bolt H M, Vercruysse A, Rogiers V
Department of Toxicology, Vrije Universiteit Brussels, Belgium.
Arch Toxicol. 2000 Dec;74(10):587-92. doi: 10.1007/s002040000176.
The collagen gel sandwich culture of hepatocytes has been proposed as one of the most suitable culture models available for biotransformation studies of xenobiotics. It is a complex model which imitates the cascade of enzymatic events of in vivo biotransformation and allows investigation of biological endpoints under realistic conditions. The biotransformation of trichloroethylene (TRI) has been studied in this model using rat hepatocytes. Headspace gas chromatographic measurements revealed that hepatocytes, cultured for 4 days in this in vitro system, metabolised TRI into the major oxidative metabolites trichloroacetic acid (TCA) and trichloroethanol (TCE). Cultured hepatocytes were exposed either to TRI, or to TCA and TCE. Endpoints studied were albumin secretion and the cytochrome P450 (CYP)-dependent enzymatic activities ethoxyresorufin O-deethylase (EROD), pentoxyresorufin O-depentylase (PROD) and N-nitrosodimethylamine demethylase (NDMA). The results show that both the parent compound and its metabolites exert specific effects on different CYP-dependent mono-oxygenase activities, as seen in vivo. It is suggested that collagen gel sandwich cultures represent a useful in vitro model for the investigation of metabolism-linked toxicity studies.
肝细胞胶原凝胶夹心培养已被认为是用于异生物质生物转化研究的最合适的培养模型之一。它是一个复杂的模型，可模拟体内生物转化的酶促事件级联反应，并能在实际条件下研究生物学终点。已在该模型中使用大鼠肝细胞研究了三氯乙烯（TRI）的生物转化。顶空气相色谱测量结果显示，在这个体外系统中培养4天的肝细胞将TRI代谢为主要的氧化代谢产物三氯乙酸（TCA）和三氯乙醇（TCE）。将培养的肝细胞暴露于TRI、TCA或TCE中。所研究的终点包括白蛋白分泌以及细胞色素P450（CYP）依赖性酶活性乙氧异吩恶唑酮O-脱乙基酶（EROD）、戊氧异吩恶唑酮O-脱戊基酶（PROD）和N-亚硝基二甲胺脱甲基酶（NDMA）。结果表明，母体化合物及其代谢产物对不同的CYP依赖性单加氧酶活性均有特定影响，这与在体内观察到的情况一致。有人提出，胶原凝胶夹心培养是用于研究代谢相关毒性研究的一种有用的体外模型。
