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培养的鲤鱼肝细胞中δ9-去饱和酶的等温诱导。

An isothermal induction of delta 9-desaturase in cultured carp hepatocytes.

作者信息

Macartney A I, Tiku P E, Cossins A R

机构信息

Department of Environmental and Evolutionary Biology, University of Liverpool, UK.

出版信息

Biochim Biophys Acta. 1996 Aug 16;1302(3):207-16. doi: 10.1016/0005-2760(96)00063-x.

Abstract

Cold exposure of carp leads to the induced activity of the hepatic delta 9-desaturase (Schünke, M. and Wodtke, E. (1983) Biochim. Biophys. Acta 734, 70-75). We have investigated the controlled expression of this enzyme using isolated carp hepatocytes. Culture at 30 degrees C, of cells isolated from 30 degrees C-acclimated carp. resulted in an 8-13-fold increase in desaturase-specific activity over 4 days, whilst another enzyme of intermediary metabolism, glucose-6-phosphatase, decreased by more than 60%. This desaturase induction was associated with a loss of intracellular lipid vesicles and with increases in the levels of oleic acid of membrane phosphoglycerides and corresponding decreases in 22:6(n - 3). Supplementation of cultures with oleic acid and with polyunsaturated fatty acids did not cause any reduction in the desaturase induction. The level of immunodetectable desaturase protein increased during culture at 30 degrees C and a desaturase mRNA was detected after 2 days of culture by Northern analysis. These results suggest that in vitro culture leads to an increased synthesis of desaturase protein by means of activated gene transcription. Significantly, transfer of cultures of 30 degrees C-acclimated hepatocytes to 10 degrees C resulted in a smaller induction of desaturase activity; thus cold transfer of cells in itself did not induce hepatocyte desaturase activity as does whole animal cooling. This suggests either that cold induction of desaturase activity in vivo involves systemic control or that the conditions imposed by culture prevent cold induction.

摘要

鲤鱼受到冷刺激会导致肝脏Δ9-去饱和酶活性增强(舒恩克,M.和沃特克,E.(1983年)《生物化学与生物物理学报》734卷,第70 - 75页)。我们利用分离出的鲤鱼肝细胞研究了这种酶的调控表达。将从适应30℃环境的鲤鱼分离出的细胞在30℃下培养,4天内去饱和酶特异性活性增加了8 - 13倍,而中间代谢的另一种酶葡萄糖-6-磷酸酶则下降了60%以上。这种去饱和酶的诱导与细胞内脂质小泡的减少以及膜磷酸甘油酯中油酸水平的增加和22:6(n - 3)相应减少有关。向培养物中添加油酸和多不饱和脂肪酸并没有导致去饱和酶诱导的任何降低。在30℃培养期间,免疫可检测的去饱和酶蛋白水平增加,并且通过Northern分析在培养2天后检测到了去饱和酶mRNA。这些结果表明,体外培养通过激活基因转录导致去饱和酶蛋白合成增加。值得注意的是,将适应30℃的肝细胞培养物转移到10℃会导致去饱和酶活性的诱导作用较小;因此,细胞的冷转移本身并不会像整个动物降温那样诱导肝细胞去饱和酶活性。这表明要么体内去饱和酶活性的冷诱导涉及全身控制,要么培养所施加的条件阻止了冷诱导。

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