Culture of rat hepatocytes in collagen sandwich system and its cytochrome P450 activity measurement / 中国药理学通报

ABSTRACT

AIMTo establish a collagen sandwich system (CSS) wit h self-prepared collagen for rat hepatocytes culture[FQ(14*2。46,X-WZ]in vitro, and study the effect of CYP3A substrates on cyt o chrome P450 activities of hepatocyte cultured by CSS. METHODSS terilized type Ⅰ collagen was prepared from rat tail tendons. The morphology of hepatocyte cultured by collagen coated (CC) and CSS method was investigated and compared. Activities of ALT, AST, and LDH in aliquots of extracellular medium w ere measured by biochemical assay. CYP1A, CYP2E1 and CYP3A activities of hepatoc ytes were also determined. RESULTSBy CSS method, the shape of hepatocytes was maintained, and the structure of cells was integrated after cult ivated for 6 d. However, in CC method after 3 d, hepatocytes dropped from the co llagen matrix, and the activities of ALT, AST and LDH in aliquots were increased . As compared with control group, in CSS cultivated hepatocytes, the activities of CYP3A increased 33%, and CYP2E1 decreased 45% by nimodipine, and CYP3A was in duced 1 94-fold (P