College of Veterinary Medicine, Northeast Agricultural University, Harbin, 150030, PR China.
College of Veterinary Medicine, Northeast Agricultural University, Harbin, 150030, PR China.
Fish Shellfish Immunol. 2023 Aug;139:108905. doi: 10.1016/j.fsi.2023.108905. Epub 2023 Jun 20.
A lack of the trace element zinc (Zn) in freshwater environments causes slow growth and malnutrition and affects the normal biological functions of organisms. In this study, a Zn deficiency model of grass carp hepatocytes was established with TPEN. Acetylcysteine (NAC) was used as an inhibitor. TPEN was added to L8824 cell culture medium, and LDH, AST, ALT, and AKP activities were enhanced in a Zn-deficient environment, leading to abnormal hepatopancreas function. Fluorescence microscopy showed an increase in ROS levels, and antioxidant enzyme activity assays revealed that SOD, CAT, GSH-PX, and T-AOC activities were decreased, indicating oxidative stress caused by Zn deficiency. The RT‒PCR results showed that the mRNA expression of GRP78, PERK, EIF2α, ATF4, and Chop was increased due to the addition of TPEN. Calcium kits showed increased Ca levels. The RT‒PCR results showed that the mRNA expression levels of Caspase-12, Caspase-9, Caspase-3, and PARP apoptotic were increased due to the addition of TPEN. RT‒PCR and ELISA showed that the expression levels of interleukin-1β (IL-1β), interleukin-8 (IL-8), tumour necrosis factor (TNF-α), and inducible nitric oxide synthase (iNOS) were increased. This led to the conclusion that Zn deficiency in the freshwater environment caused inflammation and apoptosis in hepatocytes in grass carp. For the first time, apoptosis caused by endoplasmic reticulum stress in grass carp hepatocytes due to Zn deficiency was studied in the context of Ca. The present study provided some insight into the adverse effects of Zn deficiency in freshwater environments on fish.
在淡水环境中缺乏微量元素锌（Zn）会导致生长缓慢和营养不良，并影响生物体的正常生物功能。在这项研究中，使用 TPEN 建立了草鱼肝细胞的缺锌模型。使用乙酰半胱氨酸（NAC）作为抑制剂。将 TPEN 添加到 L8824 细胞培养基中，在缺锌环境下会增强 LDH、AST、ALT 和 AKP 的活性，导致肝胰腺功能异常。荧光显微镜显示 ROS 水平升高，抗氧化酶活性测定显示 SOD、CAT、GSH-PX 和 T-AOC 活性降低，表明缺锌引起氧化应激。RT-PCR 结果显示，由于添加了 TPEN，GRP78、PERK、EIF2α、ATF4 和 Chop 的 mRNA 表达增加。钙试剂盒显示 Ca 水平增加。RT-PCR 结果显示，由于添加了 TPEN，Caspase-12、Caspase-9、Caspase-3 和 PARP 凋亡的 mRNA 表达水平增加。RT-PCR 和 ELISA 显示白细胞介素-1β（IL-1β）、白细胞介素-8（IL-8）、肿瘤坏死因子（TNF-α）和诱导型一氧化氮合酶（iNOS）的表达水平增加。这得出结论，淡水环境中缺锌会导致草鱼肝细胞发生炎症和凋亡。首次研究了 Zn 缺乏导致 Ca 草鱼肝细胞内质网应激引起的细胞凋亡。本研究为 Zn 缺乏对淡水环境中鱼类的不良影响提供了一些见解。
