College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, PR China.
College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, PR China; Key Laboratory of the Provincial Education, Department of Heilongjiang for Common Animal Disease Prevention and Treatment, PR China.
Pestic Biochem Physiol. 2023 Nov;196:105625. doi: 10.1016/j.pestbp.2023.105625. Epub 2023 Sep 22.
Cypermethrin (CYP, IUPAC name: [cyano-(3-phenoxyphenyl)methyl] 3-(2,2-dichloroethenyl)-2,2-dimethylcyclopropane-1-carboxylate) is a pyrethroid insecticide that poses a threat to the health of humans and aquatic animals due to its widespread use and environmental contamination. However, the mechanism of CYP on apoptosis, autophagy and inflammation in hepatocytes of carp (Cyprinus carpio) is unknown. We hypothesized that CYP caused damage to hepatocytes through the endoplasmic reticulum stress (ERS) pathway, CCK-8 was used to detect the toxic effects of different doses of CYP on hepatocytes, and finally low (L, 10 μM), medium (M, 40 μM), and high (H, 80 μM) doses of CYP was selected to construct the model. ROS staining, oxidative stress-related indices (MDA, CAT, T-AOC, SOD), AO/EB staining, MDC staining, and the expression levels of related genes were detected using qRT-PCR and western blot. Our results showed that CYP exposure resulted in an increase in ROS production, an increase in MDA content, and a decrease in the activity of CAT, SOD, and T-AOC in hepatocytes; the proportion of apoptotic, necrotic, and autophagic cells increased significantly in a dose-dependent manner. We also found that CYP exposure increased the expression levels of endoplasmic reticulum-related genes (GRP78, PERK, IRE-1, ATF-6 and CHOP), apoptosis (Bcl-2, Bax, Caspase-3, Caspase-9 and Cyt-c) and autophagy-related genes (LC3b, Beclin1 and P62) also showed dose-dependent changes, and the expression levels of inflammation-related genes (NF-κB, TNF-α, IL-1β, IL-6) were also significantly elevated. Thus, we demonstrated that CYP exposure caused apoptosis, autophagy and inflammation in hepatocytes via ERS-ROS-NF-κB axis. This research contributes to our understanding of the molecular mechanisms underlying CYP-induced damage in hepatocytes of carp (Cyprinus carpio).
氯菊酯（CYP，IUPAC 名：[氰基-(3-苯氧基苯基)甲基]3-(2,2-二氯乙烯基)-2,2-二甲基环丙烷-1-羧酸酯）是一种拟除虫菊酯类杀虫剂，由于其广泛使用和环境污染，对人类和水生动物的健康构成威胁。然而，氯菊酯对鲤鱼（Cyprinus carpio）肝细胞凋亡、自噬和炎症的作用机制尚不清楚。我们假设 CYP 通过内质网应激（ERS）途径对肝细胞造成损伤，用 CCK-8 检测不同剂量 CYP 对肝细胞的毒性作用，最后选择低（L，10 μM）、中（M，40 μM）和高（H，80 μM）剂量 CYP 构建模型。用 ROS 染色、氧化应激相关指标（MDA、CAT、T-AOC、SOD）、AO/EB 染色、MDC 染色和 qRT-PCR、western blot 检测相关基因的表达水平。结果表明，CYP 暴露导致肝细胞中 ROS 产生增加，MDA 含量增加，CAT、SOD 和 T-AOC 活性降低；凋亡、坏死和自噬细胞的比例明显增加，呈剂量依赖性。我们还发现，CYP 暴露增加了内质网相关基因（GRP78、PERK、IRE-1、ATF-6 和 CHOP）、凋亡（Bcl-2、Bax、Caspase-3、Caspase-9 和 Cyt-c）和自噬相关基因（LC3b、Beclin1 和 P62）的表达水平，也呈剂量依赖性变化，炎症相关基因（NF-κB、TNF-α、IL-1β、IL-6）的表达水平也明显升高。因此，我们证明 CYP 通过 ERS-ROS-NF-κB 轴导致肝细胞凋亡、自噬和炎症。这项研究有助于我们理解 CYP 对鲤鱼（Cyprinus carpio）肝细胞损伤的分子机制。
