Time course of cytochromes P450 decline during rat hepatocyte isolation and culture: effect of L-NAME
Time course of cytochromes P450 decline during rat hepatocyte isolation and culture: effect of L-NAME
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DOI：
10.1016/s0887-2333(02)00118-2
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发表时间：
2003-02-01
影响因子：
3.2
通讯作者：
Richert, L
中科院分区：
文献类型：
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作者：
Binda, D;Lasserre-Bigot, D;Richert, L
本工作描述了胶原酶灌注对肝细胞微粒体细胞色素（CYP）依赖性单加氧酶活性的同工酶相关效应：分离后的大鼠肝细胞微粒体中CYP 1A 1/2-、2B 1/2-、3A 1/2-和2 E 1-依赖性活性约为肝微粒体的60%，CYP 4A 1-依赖性活性与肝微粒体相当。相比之下，肝细胞分离不影响各种α亚型的微粒体蛋白含量。这与胶原酶消化法分离肝细胞过程中β-内酰胺酶失活的假设一致。发现L-NAME（1 mm）不能保护肝细胞分离后CYP依赖性单加氧酶活性的下降。这是可能的，谷胱甘肽过氧化物酶活性的降低观察到的L-NAME的存在下，即抑制防御过氧亚硝酸盐，可以抵消的有益效果L-NAME对一氧化氮合成抑制。目前的工作还表明，L-NAME不能避免进行性的，异构体特异性的，最可能的翻转相关的，在培养的肝细胞中的α-淀粉酶蛋白和相关的单加氧酶活性的下降。在大鼠肝细胞培养物中，目前未知但不依赖于一氧化氮，因此，在培养的大鼠肝细胞中似乎发生了调节障碍。(C)2002爱思唯尔科技有限公司。保留所有权利。
The present work describes an isozyme-related effect of collagenase perfusion on hepatocyte microsomal cytochrome (CYP)dependent monooxygenase activities: CYP 1A1/2-, 2B1/2-, 3A1/2- and 2E1-dependent activities in microsomes from rat hepatocytes after isolation were about 60% of that of liver microsomes, and CYP 4A1-dependent activity was equivalent to liver microsomes. In contrast, the microsomal protein content of the various CYP isoforms was not affected by hepatocyte isolation. This is in accordance with the hypothesis of CYP inactivation during the process of hepatocyte isolation by collagenase digestion. L-NAME (I mm) was found unable to protect from the decline of CYP-dependent monooxygenase activities following hepatocyte isolation. It is possible that the decrease in glutathione peroxidase activity observed in the presence Of L-NAME, namely depression of defense against peroxynitrite, could counteract the beneficial effect Of L-NAME on nitric oxide synthesis inhibition. The present work also shows that L-NAME could not avoid the progressive, isoform-specific, most probably turn over-related, decline of CYP proteins and related monooxygenase activities in cultured hepatocytes. Dysregulations in the mechanisms of CYP expression in rat hepatocyte cultures, presently unknown but nitric oxide independent, thus appear to occur in cultured rat hepatocytes. (C) 2002 Elsevier Science Ltd. All rights reserved.