College of Veterinary Medicine, Northeast Agricultural University, Harbin, 150030, Peoples R China.
College of Life Sciences, Northeast Agricultural University, Harbin, 150030, Peoples R China.
Fish Shellfish Immunol. 2024 Nov;154:109944. doi: 10.1016/j.fsi.2024.109944. Epub 2024 Oct 5.
Cypermethrin is a toxic pesticide that has infiltrated water bodies due to its widespread use. This contamination has led to detrimental effects on the immune organs of aquatic species, including fish. The natural fat-soluble orange-red carotenoid, astaxanthin (MAT), derived from microalgae, possesses anti-inflammatory, antioxidant, and immunomodulatory properties. To elucidate the mechanism of CY induced damage to carp liver cells and assess the potential protective effects of MAT, we established a carp hepatocyte model exposed to CY and/or MAT. Hepatocytes from carp (Cyprinus carpio) were treated with either 8 μM CY or 60 μM MAT for 24 h. Upon exposure CY, a significant increase in reactive oxygen species (ROS) was observed alongside a diminution in the activities of key antioxidant enzymes, superoxide dismutase (SOD) and catalase (CAT), suggesting an impairment of cellular antioxidant capacity. Subsequently, acridine orange/ethidium bromide (AO/EB) staining and flow cytometry analysis revealed that hepatocytes exposed to CY exhibited a higher incidence of necroptosis, associated with an elevated mitochondrial Ca concentration, which contributed to cellular dysfunction. Furthermore, exposure to CY also activated the ROS-NF-κB-RIPK3/MLKL signaling pathway, increasing the levels of necroptosis-related regulatory factors (RIP1, RIP3, and MLKL) in hepatocytes and the expression of inflammatory genes (IL-6, IFN-γ, IL-4, IL-1β, and TNF-α), which led to immune dysfunction in hepatocytes. The immunotoxic effects induced by CY were mitigated by MAT treatment, suggesting its potential in alleviating the aforementioned changes caused by CY. Overall, the data suggested that MAT therapy could enhance hepatocyte defenses against CY-induced necroptosis and inflammatory responses by regulating mitochondrial Ca homeostasis and inhibiting the ROS-NF-κB-RIPK3/MLKL signaling cascade. This study elucidated the potential benefits of employing MAT to protect farmed fish from agrobiological hazards during CY exposure, underscoring the practical applications of MAT in aquaculture.
氯菊酯是一种毒性农药，由于其广泛使用而渗透到水体中。这种污染对鱼类等水生物种的免疫器官造成了有害影响。天然脂溶性橙红色类胡萝卜素虾青素（MAT）来源于微藻，具有抗炎、抗氧化和免疫调节作用。为了阐明 CY 诱导鲤鱼肝细胞损伤的机制，并评估 MAT 的潜在保护作用，我们建立了暴露于 CY 和/或 MAT 的鲤鱼肝细胞模型。用 8μM CY 或 60μM MAT 处理鲤鱼（Cyprinus carpio）肝细胞 24 小时。暴露于 CY 后，观察到活性氧（ROS）显著增加，关键抗氧化酶超氧化物歧化酶（SOD）和过氧化氢酶（CAT）的活性降低，表明细胞抗氧化能力受损。随后，吖啶橙/溴化乙锭（AO/EB）染色和流式细胞术分析显示，暴露于 CY 的肝细胞发生坏死性凋亡的发生率更高，与线粒体 Ca 浓度升高有关，这导致细胞功能障碍。此外，暴露于 CY 还激活了 ROS-NF-κB-RIPK3/MLKL 信号通路，增加了肝细胞中坏死性凋亡相关调节因子（RIP1、RIP3 和 MLKL）的水平以及炎症基因（IL-6、IFN-γ、IL-4、IL-1β 和 TNF-α）的表达，导致肝细胞免疫功能障碍。MAT 处理减轻了 CY 引起的免疫毒性作用，表明其在缓解 CY 引起的上述变化方面具有潜力。总的来说，数据表明，MAT 治疗通过调节线粒体 Ca 稳态和抑制 ROS-NF-κB-RIPK3/MLKL 信号通路，增强了肝细胞对 CY 诱导的坏死性凋亡和炎症反应的防御能力。这项研究阐明了在 CY 暴露期间使用 MAT 来保护养殖鱼类免受农业生物危害的潜在益处，强调了 MAT 在水产养殖中的实际应用。
