锌、NAC对镉染毒巨噬细胞的保护作用

Protective effects of zinc and NAC on cadmium-exposed macrophage

探讨镉(Cd)对家禽巨噬细胞的毒性损伤和免疫功能的影响,以及锌、N-乙酰半胱氨酸(NAC)对Cd染毒巨噬细胞的保护作用。试验建立Cd^(2+)(20,50μmol/L)染毒HD-11细胞模型,研究不同时间段(6,12 h)Cd对HD-11细胞毒性损伤,并通过向细胞培养基中补充Zn^(2+)(20μmol/L)、NAC(500μmol/L)的方式,探讨Zn^(2+)和NAC对Cd染毒巨噬细胞细胞骨架、细胞凋亡、ROS产生、线粒体膜电位(MMP)水平及免疫功能的影响。通过检测细胞金属元素结合原件(MTF-1)、金属硫蛋白(MT-1)基因转录水平,揭示Zn^(2+)和NAC对Cd染毒巨噬细胞的调控机理。结果显示,Cd染毒造成HD-11细胞骨架蛋白降解、ROS产生、MMP水平下降、细胞凋亡增加以及IL-1β、IL-8、IL-10基因表达量降低。Zn^(2+)、NAC添加能不同程度地降低Cd染毒巨噬细胞结构损伤,减少ROS产生和MMP去极化水平,增强细胞抗氧化能力和免疫功能,促进MTF-1和MT-1基因表达。结果表明,Cd染毒可以造成鸡巨噬细胞浓度—时间依赖性结构损伤和免疫功能紊乱,单独或联合添加Zn^(2+)、NAC可通过增强细胞抗氧化水平,有效抑制Cd染毒鸡巨噬细胞毒性损伤,改善细胞免疫功能。

The objective of this study is to investigate the effects of cadmium induced cytotoxicity,immune adjustment on avian macrophage,the protective effects of zinc and NAC on cadmium-exposed macrophage.In this experiment,cadmium poisoning(20,50μmol/L)cell model was established.Cadmium toxicity damage to HD-11 cells was investigated at different time periods(6,12 h).Additionally,the effects of Zn^(2+)and NAC on cytoskeleton,apoptosis rate,reactive oxygen species(ROS)content,level of mitochondrial membrane potential(MMP)of cadmium induced macrophages by supplementing the cell culture medium with Zn^(2+)(20μmol/L),NAC(500μmol/L)was investigated.The regulatory mechanisms of Zn^(2+)and NAC on cadmium-stained macrophages were revealed by detecting the transcript levels of cellular metal-element binding protomer(MTF-1)and metallothionein(MT-1)genes.The results showed that cadmium poisoning caused the degradation of HD-11 cytoskeletal proteins,ROS production,decrease of MMP level,increase of apoptosis,and decrease of IL-1β,IL-8,and IL-10 genes expression,and the addition of Zn^(2+)and NAC reduced the structural damage of cadmium-poisoned macrophages to different degrees,decreased the production of ROS and the depolarization level of MMP,and strengthened the cellular antioxidant capacity and immune function,as well as promoted the transcription levels of MTF-1 and MT-1,and the regulation of MTF-1 and MT-1 genes.and promoted MTF-1 and MT-1 gene expression.The results showed that cadmium contamination could cause concentration-time-dependent structural damage and immune dysfunction in chicken macrophages,and the addition of Zn^(2+)and NAC alone or in combination could effectively inhibit the toxicity damage of cadmiumcontaminated chicken macrophages and improve the cellular immune function by enhancing the cellular antioxidant level.