In Vitro Toxicity of Copper, Cadmium, and Chromium to Isolated Hepatocytes from Carp, Cyprinus carpio L.
In Vitro Toxicity of Copper, Cadmium, and Chromium to Isolated Hepatocytes from Carp, Cyprinus carpio L.
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DOI：
10.1007/s00128-005-0802-0
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发表时间：
2005-10
影响因子：
2.7
通讯作者：
M. Nawaz;C. Manzl;G. Krumschnabel
中科院分区：
文献类型：
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作者：
M. Nawaz;C. Manzl;G. Krumschnabel
材料与方法本实验采用急性毒性实验方法，研究了Cu、Cd和Cr（VI）对细胞的毒性作用，并对与金属诱导的细胞死亡有关的几个参数进行了研究，包括ROS的形成、脂质过氧化和细胞内游离Ca ~（2+）浓度。此外，通过观察细胞骨架的两种主要蛋白质，肌动蛋白和微管，研究了对细胞完整性的毒性作用。牛血清白蛋白（BSA）、胎牛血清（FCS）和胶原酶（VIII型）购自Sigma。Alexa荧光鬼笔环肽488、二氯荧光素二乙酸酯（DCF-DA）和Fura 2-AM来自Molecular Probes。小鼠抗B-微管蛋白单克隆抗体得自Chernicon，TRITC标记的抗小鼠抗体得自Dako。CytoTox-ONETM测定购自Promega。所有其他化学品均从当地供应商处获得。Cu、Cd和Cr（VI）由Merck（德国）公司提供，并以在蒸馏水中制备的储备溶液中的CuCl 2 - 2 H 2 O、CdCl 2和KZC 2 O 7形式施用。
MATERIALS AND METHODSThe acute cytotoxicity of Cu, Cd and Cr (VI) was evaluated and several parameters known to be involved in metal-induced cell death were investigated, including formation of ROS, lipid peroxidation, and intracellular free Ca2+(Ca2+ i). Furthermore, toxic effects exerted on cellular integrity were investigated by visualisation of two major proteins of the cytoskeleton, actin filaments and microtubules.Bovine serum albumin (BSA), fetal calf serum (FCS), and collagenase (Type VIII) were purchased from Sigma. Alexa fluor phalloidin 488, dichlorofluorescein diacetate (DCF-DA), and Fura 2-AM were from Molecular Probes. Mouse anti B-tubulin monoclonal antibody was obtained from Chernicon and TRITC-labelled anti-mouse antibody was from Dako. CytoTox-ONETM assay was purchased from Promega. All other chemicals were obtained from local suppliers. Cu, Cd and Cr (VI) were fi'orn Merck (Germany) and were applied as CuCl;> _-2H; O, CdCl2 and KZCQO7 from stock solution prepared in distilled water.