Department of Environmental Science, Baylor University, Waco, TX 76706, USA.
Department of Environmental Science, Baylor University, Waco, TX 76706, USA.
Comp Biochem Physiol C Toxicol Pharmacol. 2018 Apr;206-207:32-40. doi: 10.1016/j.cbpc.2018.02.006. Epub 2018 Feb 27.
The use of fish cell cultures has proven to be an effective tool in the study of environmental and aquatic toxicology. Valuable information can be obtained from comparisons between cell lines from different species and organs. In the present study, specific chemicals were used and biomarkers (e.g. 7-Ethoxyresorufin-O-deethylase (EROD) activity and reactive oxygen species (ROS)) were measured to assess the metabolic capabilities and cytotoxicity of the fish hepatic cell lines Hepa-E1 and RTH-149, and the fish gill cell lines RTgill-W1 and G1B. These cell lines were exposed to β-naphthoflavone (BNF) and benzo[a]pyrene (BaP), the pharmaceutical tamoxifen (TMX), and the organic peroxide tert-butylhydroperoxide (tBHP). Cytotoxicity in gill cell lines was significantly higher than in hepatic cells, with BNF and TMX being the most toxic compounds. CYP1-like associated activity, measured through EROD activity, was only detected in hepatic cells; Hepa-E1 cells showed the highest activity after exposure to both BNF and BaP. Significantly higher levels of CYP3A-like activity were also observed in Hepa-E1 cells exposed to TMX, while gill cell lines presented the lowest levels. Measurements of ROS and antioxidant enzymes indicated that peroxide levels were higher in gill cell lines in general. However, levels of superoxide were significantly higher in RTH-149 cells, where no distinctive increase of superoxide-related antioxidants was observed. The present study demonstrates the importance of selecting adequate cell lines in measuring specific metabolic parameters and provides strong evidence for the fish hepatocarcinoma Hepa-E1 cells to be an excellent alternative in assessing metabolism of xenobiotics, and in expanding the applicability of fish cell lines for in vitro studies.
鱼类细胞培养已被证明是研究环境和水生毒理学的有效工具。通过比较不同物种和器官的细胞系，可以获得有价值的信息。在本研究中，使用了特定的化学物质，并测量了生物标志物（例如 7-乙氧基香豆素-O-脱乙基酶（EROD）活性和活性氧物种（ROS）），以评估鱼类肝细胞系 Hepa-E1 和 RTH-149 以及鱼类鳃细胞系 RTgill-W1 和 G1B 的代谢能力和细胞毒性。这些细胞系暴露于β-萘黄酮（BNF）和苯并[a]芘（BaP）、药物他莫昔芬（TMX）和有机过氧化物叔丁基过氧化物（tBHP）。鳃细胞系的细胞毒性明显高于肝细胞系，BNF 和 TMX 是最具毒性的化合物。通过 EROD 活性测量的 CYP1 样相关活性仅在肝细胞系中检测到；暴露于 BNF 和 BaP 后，Hepa-E1 细胞表现出最高的活性。在暴露于 TMX 的 Hepa-E1 细胞中还观察到 CYP3A 样活性显著升高，而鳃细胞系则表现出最低的活性。ROS 和抗氧化酶的测量表明，一般来说，鳃细胞系中的过氧化物水平较高。然而，在 RTH-149 细胞中，超氧化物水平显著升高，但没有观察到与超氧化物相关的抗氧化剂明显增加。本研究表明，在测量特定代谢参数时选择适当的细胞系非常重要，并为使用鱼类肝癌细胞系 Hepa-E1 评估外源物质代谢提供了有力证据，同时也为鱼类细胞系在体外研究中的应用提供了更广泛的可能性。
