Department of Pharmacology and Toxicology, Faculty of Veterinary Medicine, Universidad Complutense de Madrid, 28040, Madrid, Spain.
Environ Res. 2019 Oct;177:108579. doi: 10.1016/j.envres.2019.108579. Epub 2019 Jul 12.
This study was designed to assess oxidative stress induction in human neuroblastoma SH-SY5Y cells in response to cyfluthrin exposure. Cell viability MTT assay was carried out to assess cyfluthrin cytotoxicity; IC and IC values for cyfluthrin were calculated to be 4.81 ± 0.92 μM and 19.39 ± 3.44 μM, respectively. Cyfluthrin induced a significant increase in ROS generation, lipid peroxides measured as malondialdehyde (MDA) and nitric oxide (NO) production and a significant decrease in NQO1 activity. The antioxidant activity of melatonin (MEL), Trolox, N-acetylcysteine (NAC) and Sylibin against cyfluthrin-induced oxidative stress was examined. Cyfluthrin increased significantly gene expressions of apoptosis, proinflammation and oxidative stress (Bax, Bcl-2, Casp-3, BNIP3, AKT1, p53, APAF1, NFκB1, TNFα and Nrf2) mediators. In the most genes, the mRNA levels induced by cyfluthrin were partially reduced by MEL (1 μM). Cyfluthrin effects on gene expression profiling of oxidative stress pathway by Real-Time PCR array analysis showed that of the 84 genes examined, (fold change > 1.5) changes in mRNA levels were detected in 31 genes: 13 upregulated and 18 down-regulated. A fold change >3.0 fold was observed on upregulated CYBB, DUOX1, DUOX2, AOX1, BNIP3, HSPA1A, NOS2, and NQO1 genes. The greater fold change reversion (2.5 fold) by MEL (1 μM) was observed on cyfluthrin-upregulated CYBB, AOX1, BNIP3 and NOS2 genes. These results demonstrated that oxidative stress is a key element in cyfluthrin induced neurotoxicity as well as MEL may play a role in reducing cyfluthrin-induced oxidative stress.
本研究旨在评估氯氟氰菊酯暴露对人神经母细胞瘤 SH-SY5Y 细胞氧化应激的诱导作用。通过 MTT 法检测细胞活力来评估氯氟氰菊酯的细胞毒性；计算得出氯氟氰菊酯的 IC 和 IC 值分别为 4.81±0.92 μM 和 19.39±3.44 μM。氯氟氰菊酯诱导 ROS 生成、丙二醛（MDA）和一氧化氮（NO）生成显著增加，NQO1 活性显著降低。研究了褪黑素（MEL）、Trolox、N-乙酰半胱氨酸（NAC）和水飞蓟宾对氯氟氰菊酯诱导的氧化应激的抗氧化活性。氯氟氰菊酯显著增加了凋亡、促炎和氧化应激基因表达（Bax、Bcl-2、Casp-3、BNIP3、AKT1、p53、APAF1、NFκB1、TNFα和 Nrf2）介质。在大多数基因中，MEL（1 μM）部分降低了氯氟氰菊酯诱导的 mRNA 水平。氯氟氰菊酯对实时 PCR 阵列分析氧化应激途径基因表达谱的影响表明，在所研究的 84 个基因中，有 31 个基因的 mRNA 水平发生了变化（fold change >1.5）：13 个上调，18 个下调。上调基因有 CYBB、DUOX1、DUOX2、AOX1、BNIP3、HSPA1A、NOS2 和 NQO1，fold change >3.0 倍。MEL（1 μM）对氯氟氰菊酯上调的 CYBB、AOX1、BNIP3 和 NOS2 基因的逆转倍数更高（2.5 倍）。这些结果表明，氧化应激是氯氟氰菊酯诱导神经毒性的关键因素，而 MEL 可能在减轻氯氟氰菊酯诱导的氧化应激中发挥作用。
