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石油水溶性成分对甲壳动物博氏沼虾脂质分解代谢的激活作用。

Activation of lipid catabolism by the water-soluble fraction of petroleum in the crustacean Macrobrachium borellii.

作者信息

Lavarías S, Pollero R J, Heras H

机构信息

Instituto de Investigaciones Bioquímicas de La Plata (INIBIOLP), CONICET-Cátedra de Bioquímica, UNLP, Argentina.

出版信息

Aquat Toxicol. 2006 May 1;77(2):190-6. doi: 10.1016/j.aquatox.2005.12.002. Epub 2006 Jan 18.

Abstract

Little is known about the effect of the water-soluble fraction of crude oil (WSF) on lipid metabolism in invertebrates. The effect of the WSF on the triacylglycerol (TAG) mobilization, fatty acid activation and degradation was evaluated in the decapod Macrobrachium borellii, exposing adult and eggs at different stages of development for 7 days to a sublethal concentration of WSF. Using radioactive tracers, mitochondrial palmitoyl-CoA synthetase (ACS), triacylglycerol lipase (TAG-lipase) and fatty acid beta-oxidation system activities were assayed. Before studying the effect of WSF, the kinetic parameters of ACS were determined in purified mitochondria. Its optimal temperature and pH were 32 degrees C and 8.0, respectively, the apparent K(m) 2.48 micromol l(-1), and its V(max) of 1.93 nmol min(-1) mg protein(-1). These kinetic parameters differed significantly from this shrimp's microsomal isoform. After 7 days exposure to a sublethal concentration of WSF (0.6 mg/l), changes were observed in the enzymatic activity of all enzymes or enzymatic system assayed in adult midgut gland as well as in stage 5 eggs, a period of active organogenesis. An increase in the mobilization of energy stores was detected as early as stage 4, where TAG-lipase activity increased by 27% in exposed eggs. The increase was even more marked in exposed eggs at stage 5 where a three-fold rise (154%) was determined. Exposed adult shrimp also showed an augmented lipase activity by 38%. Fatty acid beta-oxidation increased by 51.0 and 35.5% in midgut gland and eggs at stage 5, respectively, but no changes were observed at less-developed stages. Mitochondrial fatty acid activation by ACS also increased in adults and stage 5 eggs by 7.4 and 52.0%, respectively. A similar response of the lipid catabolic pathways to WSF contamination in both adult and eggs, suggests that the exposure to this pollutant causes an increase in the energy needs of this shrimp. When validated by field studies, these catabolic enzymes could be employed as early pollution biomarkers.

摘要

关于原油水溶性部分(WSF)对无脊椎动物脂质代谢的影响,目前所知甚少。在十足目动物博氏沼虾中评估了WSF对三酰甘油(TAG)动员、脂肪酸活化和降解的影响,将成年个体和处于不同发育阶段的卵暴露于亚致死浓度的WSF中7天。使用放射性示踪剂,测定了线粒体棕榈酰辅酶A合成酶(ACS)、三酰甘油脂肪酶(TAG-脂肪酶)和脂肪酸β-氧化系统的活性。在研究WSF的影响之前,在纯化的线粒体中测定了ACS的动力学参数。其最适温度和pH分别为32℃和8.0,表观K(m)为2.48 μmol l(-1),V(max)为1.93 nmol min(-1) mg蛋白(-1)。这些动力学参数与该虾的微粒体同工型有显著差异。在暴露于亚致死浓度的WSF(0.6 mg/l)7天后,在成年中肠腺以及处于活跃器官发生期的第5阶段卵中,所检测的所有酶或酶系统的酶活性均出现了变化。早在第4阶段就检测到能量储存动员增加,暴露卵中的TAG-脂肪酶活性增加了27%。在第5阶段的暴露卵中增加更为明显,增加了两倍(154%)。暴露的成年虾脂肪酶活性也增加了38%。在第5阶段,中肠腺和卵中的脂肪酸β-氧化分别增加了51.0%和35.5%,但在发育程度较低的阶段未观察到变化。成年个体和第5阶段卵中线粒体脂肪酸由ACS活化的情况也分别增加了7.4%和52.0%。成年个体和卵中脂质分解途径对WSF污染的类似反应表明,暴露于这种污染物会导致该虾的能量需求增加。当通过实地研究验证后,这些分解代谢酶可作为早期污染生物标志物。

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