Time-dependent production of nitric oxide by rat hepatocyte suspensions.
Time-dependent production of nitric oxide by rat hepatocyte suspensions.
复制标题
大鼠肝细胞悬浮液产生的一氧化氮的时间依赖性。
DOI：
10.1016/s0006-2952(99)00053-2
复制
发表时间：
1999
影响因子：
5.8
通讯作者：
Fariss,MW
中科院分区：
文献类型：
--
作者：
Nicholls-Grzemski,FA;Tirmenstein,MA;Fariss,MW
通过胶原酶灌流制备的分离的肝细胞悬浮液在8小时的孵育过程中将高水平的亚硝酸盐释放到细胞外培养液中。释放是时间依赖的，增加首先发生在4小时，并持续整个潜伏期的剩余时间。一氧化氮合酶抑制剂氨基胍和N-硝基-L-精氨酸甲酯(L-NAME)抑制了亚硝酸盐的生成，表明亚硝酸盐来源于一氧化氮合酶活性产生的一氧化氮(NO)。亚硝酸盐的产生与细菌或库普弗细胞污染无关。蛋白质合成抑制剂放线菌胺和转录抑制剂放线菌素D也能抑制实质肝细胞亚硝酸盐的产生。钙非依赖性一氧化氮合酶活性随着孵育时间的增加而增加，这种增加与观察到的亚硝酸盐产生的增加是一致的。我们的结果表明，在分离肝细胞后，一氧化氮合酶被诱导，这种诱导导致了高水平的NO的形成。
Isolated hepatocyte suspensions prepared by collagenase perfusion released high levels of nitrite into the extracellular medium during an 8-hr incubation. The release was time dependent, with increases first occurring by 4 hr and continuing throughout the remainder of the incubation period. Nitrite production was inhibited by the nitric oxide synthase (NOS) inhibitors aminoguanidine and NG-nitro-l-arginine methyl ester (l-NAME), indicating that the nitrite is derived from nitric oxide (NO) production from NOS activity. Nitrite production was not related to bacterial or Kupffer cell contamination. The protein synthesis inhibitor cycloheximide and the transcription inhibitor actinomycin D also prevented nitrite production by parenchymal hepatocytes. Calcium-independent NOS enzyme activity increased with incubation times, and this increase coincided with the observed increases in nitrite production. Our results suggest that NOS is induced following the isolation of hepatocytes, and this induction results in the formation of high levels of NO.