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舒张压对鲤鱼鳃和肝脏组织的组织学及热休克蛋白70表达的影响

Impact of DBP on histology and expression of HSP 70 in gill and liver tissue of Cyprinus carpio.

作者信息

Agus Hizlan H, Erkmen Belda, Sümer Sibel, Sepici-Dinçel Aylin, Erkoç Figen

机构信息

Department of Biology, Hacettepe University, Beytepe, 06800, Ankara, Turkey,

出版信息

Mol Biol Rep. 2015 Sep;42(9):1409-17. doi: 10.1007/s11033-015-3920-8. Epub 2015 Aug 27.

Abstract

Di-n-butyl phthalate (DBP) widely used plasticizer in the plastic industry, affects regulation of the endocrine system and causes toxicity in animals. In the present study, the aim was to study the toxic effects/damages of DBP exposure using Hsp70 levels and histopathological changes in Carp liver and gill. Hsp70 expression levels were assessed as specific biomarker of in vivo ecotoxicological stress. Carp (Cyprinus carpio) were exposed to sub-lethal concentration of DBP (di-n-butyl phthalate, 1 mg/L) for 4, 24 and 96 h. Gill and liver tissues were evaluated histopathologically and RNA quantifications for Hsp70 expression levels were carried out using a two-step real-time RT-PCR. In liver, a rapid but non-significant increase in mRNA levels in the first 4 h was observed. mRNA levels significantly increased up to 2-3 fold after 24 and 96 h (p < 0.05). However, irregular mRNA level changes were also recorded: Gill specific and time-dependent regulation of Hsp70 expression were 4-5 fold inhibition after 4 and 24 h (p < 0.05), then increased up to 4 fold after 96 h (p < 0.05). Histopathological findings support altered transcription results as: Epithelial lifting, hyperplasia, fusion of secondary lamellae, telangiectasis, passive hyperemia and hydropic degeneration. Significant alterations of Hsp70 levels were likely due to a tissue specific response against chemical stress, cellular damage and lesions due to DBP. Carp was found to be a suitable experimental model for toxicology, and Hsp70 mRNA levels are reliable, specific biomarkers.

摘要

邻苯二甲酸二丁酯(DBP)是塑料工业中广泛使用的增塑剂,会影响内分泌系统的调节并对动物产生毒性。在本研究中,目的是利用鲤鱼肝脏和鳃中的热休克蛋白70(Hsp70)水平及组织病理学变化来研究DBP暴露的毒性作用/损害。Hsp70表达水平被评估为体内生态毒理学应激的特异性生物标志物。将鲤鱼(Cyprinus carpio)暴露于亚致死浓度的DBP(邻苯二甲酸二丁酯,1 mg/L)中4、24和96小时。对鳃和肝脏组织进行组织病理学评估,并使用两步实时逆转录聚合酶链反应(RT-PCR)对Hsp70表达水平进行RNA定量分析。在肝脏中,观察到在最初4小时内mRNA水平迅速但无显著增加。在24和96小时后,mRNA水平显著增加至2 - 3倍(p < 0.05)。然而,也记录到了不规则的mRNA水平变化:鳃中Hsp70表达的特异性和时间依赖性调节在4和24小时后为4 - 5倍抑制(p < 0.05),然后在96小时后增加至4倍(p < 0.05)。组织病理学结果支持转录结果的改变,表现为:上皮隆起、增生、次生鳃小片融合、毛细血管扩张、被动性充血和水样变性。Hsp70水平的显著改变可能是由于针对化学应激、细胞损伤和DBP引起的病变的组织特异性反应。鲤鱼被发现是毒理学的合适实验模型,并且Hsp70 mRNA水平是可靠的特异性生物标志物。

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