Cai ZhaoLong, Wang HongRu, Han Fuxin, Wang JingJing, Lv Hongli, Gao Xue-Jiao, Guo Meng-Yao
College of Veterinary Medicine, Northeast Agricultural University, Harbin, 150030, People's Republic of China.
Biol Trace Elem Res. 2025 Jun 24. doi: 10.1007/s12011-025-04715-w.
Zinc deficiency is closely related to oxidative stress, inflammation, and programmed cell death. In this study, mouse models with normal zinc (Con), zinc deficiency (L-Zn), and high zinc (H-Zn) and an in vitro model of AML-12 hepatocytes were established to systematically explore the effects of zinc deficiency on hepatic oxidative stress, inflammation, and programmed cell death. In vivo experiments showed that zinc deficiency significantly increased the levels of reactive oxygen species (ROS) and malondialdehyde (MDA) in the liver (P < 0.05), inhibited the activities of antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH), and induced hepatocyte edema, inflammatory infiltration, and an increase in the number of TUNEL-positive apoptotic cells. Using quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot, it was found that zinc deficiency activated the NF-κB pathway (increased expression of p-IκB α and p-NF κB p65), significantly increased the expression of pro-inflammatory factors (TNF-α, IL-6, IL-1β, IL-18), and decreased the anti-inflammatory factor IL-10. In addition, zinc deficiency upregulated apoptosis-related genes (Cyt-C, Bax, Bcl-2, Caspase-3/9), necroptosis marker indicators (RIPK1, RIPK3, MLKL), and key molecules of pyroptosis (NLRP3, ASC, GSDMD, Caspase-1), indicating the programmed cell death is activated in a number of ways. In vitro experiments further verified the above experimental results. Flow cytometry showed that the apoptosis rate of AML-12 cells in the L-Zn group (17.20%) was significantly higher than that in the Con group (4.75%) and the H-Zn group (2.55%). The experiment concluded that zinc supplementation could effectively alleviate oxidative damage, inhibit the inflammatory pathway, and reduce the expression of programmed cell death-related factors. This study confirms that zinc deficiency activates programmed cell death through a multimodal pattern and provides a theoretical basis for studies related to zinc intake imbalance leading to liver injury.
锌缺乏与氧化应激、炎症和程序性细胞死亡密切相关。在本研究中，建立了正常锌（Con）、锌缺乏（L-Zn）和高锌（H-Zn）的小鼠模型以及AML-12肝细胞的体外模型，以系统地探讨锌缺乏对肝脏氧化应激、炎症和程序性细胞死亡的影响。体内实验表明，锌缺乏显著增加了肝脏中活性氧（ROS）和丙二醛（MDA）的水平（P < 0.05），抑制了超氧化物歧化酶（SOD）、过氧化氢酶（CAT）和谷胱甘肽（GSH）等抗氧化酶的活性，并诱导肝细胞水肿、炎症浸润以及TUNEL阳性凋亡细胞数量增加。通过定量实时聚合酶链反应（qRT-PCR）和蛋白质免疫印迹法发现，锌缺乏激活了NF-κB通路（p-IκBα和p-NFκB p65表达增加），显著增加了促炎因子（TNF-α、IL-6、IL-1β、IL-18）的表达，并降低了抗炎因子IL-10。此外，锌缺乏上调了凋亡相关基因（Cyt-C、Bax、Bcl-2、Caspase-3/9）、坏死性凋亡标志物指标（RIPK1、RIPK3、MLKL）和焦亡关键分子（NLRP3、ASC、GSDMD、Caspase-1），表明程序性细胞死亡通过多种方式被激活。体外实验进一步验证了上述实验结果。流式细胞术显示，L-Zn组AML-12细胞的凋亡率（17.20%）显著高于Con组（4.75%）和H-Zn组（2.55%）。实验得出结论，补充锌可以有效减轻氧化损伤，抑制炎症通路，并降低程序性细胞死亡相关因子的表达。本研究证实锌缺乏通过多模式激活程序性细胞死亡，为锌摄入失衡导致肝损伤的相关研究提供了理论依据。
