Vandenberghe Y, Ratanasavanh D, Glaise D, Guillouzo A
INSERM, Unité de Recherches Hépatologiques U 49, Hôpital de Pontchaillou, Rennes, France.
In Vitro Cell Dev Biol. 1988 Apr;24(4):281-8. doi: 10.1007/BF02628828.
Glutathione S-transferase (GST) activity was measured in adult rat hepatocytes during either pure culture or coculture with another rat liver cell type in various media. Addition of nicotinamide, selenium, or dimethylsulfoxide, deprivation of cyst(e)ine and the use of two complex media were tested. Whatever the conditions used, after a constant decrease during the first 24 h, GST remained active over the whole culture period (1-2 wk). However, various patterns were observed: GST activity either remained relatively stable to approximately 50% of the initial value or showed a moderate or strong increase. The highest values were found in pure hepatocyte cultures maintained in the presence of nicotinamide or dimethylsulfoxide. Similar changes were observed using 1-chloro-2,4-dinitrobenzene or 1,2-dichloro-4-nitrobenzene as substrates for GST. Addition of 10(-4) M indomethacin resulted in 37 to 60% inhibition of enzyme activity. Thus, these results demonstrate that GST remained expressed during culture but its levels markedly varied depending on the medium composition and type and age of culture.
在不同培养基中，对成年大鼠肝细胞进行纯培养或与另一种大鼠肝细胞类型共培养时，测定了谷胱甘肽S-转移酶（GST）的活性。测试了添加烟酰胺、硒或二甲基亚砜、剥夺胱氨酸以及使用两种复合培养基的情况。无论使用何种条件，在最初的24小时内GST活性持续下降后，在整个培养期（1 - 2周）内仍保持活性。然而，观察到了各种模式：GST活性要么相对稳定在初始值的约50%，要么呈现中度或强烈增加。在添加烟酰胺或二甲基亚砜的纯肝细胞培养物中发现了最高值。使用1-氯-2,4-二硝基苯或1,2-二氯-4-硝基苯作为GST的底物时，观察到了类似的变化。添加10^(-4) M吲哚美辛导致酶活性受到37%至60%的抑制。因此，这些结果表明，GST在培养过程中仍有表达，但其水平根据培养基组成、培养类型和培养时间而显著变化。
