Jiangxi Provincial Key Laboratory for Animal Health, Institute of Animal Population Health, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang 330045, PR China.
School of Information Technology, Jiangxi University of Finance and Economics, Nanchang 330032, Jiangxi, PR China.
Ecotoxicol Environ Saf. 2021 Jan 15;208:111528. doi: 10.1016/j.ecoenv.2020.111528. Epub 2020 Nov 3.
Excess molybdenum (Mo) is harmful to the body, and the kidney is the vital target organ for Mo exposure. This study focused on the impacts of excess Mo on pyroptosis and the relationship between pyroptosis and apoptosis in kidney.
The duck renal tubular epithelial cells were treated with (NH)MoO·4HO (0, 480, 720 and 960 μM Mo), N-acetyl-L-cysteine (NAC) (100 μM), Z-YVAD-fluoromethylketone (YVAD) (10 μM) and the combination of Mo and NAC or YVAD for 12 h. The LDH release and IL-1β, IL-18 contents of cell supernatant were detected by LDH and ELISA kits. The MMP and ROS level were measured using MMP and ROS kits by flow cytometry. The apoptotic rate of cell was detected by AO/EB counterstaining. Pyroptosis and apoptosis-related factors mRNA and protein levels were assayed by real-time qPCR and western blot, respectively.
Excessive Mo markedly increased LDH, IL-18, IL-1β releases and induced overproduction of ROS, pyroptosis-related factors mRNA and protein levels. NAC and YVAD dramatically decreased pyroptosis induced by Mo. Simultaneously, YVAD significantly changed apoptosis-related factors mRNA and protein levels, and reduced cell apoptotic rate.
Excessive Mo exposure can induce pyroptosis by the ROS/NLRP3/Caspase-1 pathway in duck renal tubular epithelial cells, and restraining pyroptosis of Caspase-1 dependence might weaken excess Mo-induced apoptosis. The study provides theoretical basis for excess Mo exposure nephrotoxic researches on waterfowl and the interplay between pyroptosis and apoptosis highlights a new sight into the mechanism of Mo-induced nephrotoxicity.
过量的钼（Mo）对人体有害，肾脏是钼暴露的重要靶器官。本研究主要关注过量 Mo 对细胞焦亡的影响以及细胞焦亡与凋亡之间的关系。
用（NH 4 ） 2 MoO 4 ·4H 2 O（0、480、720 和 960 μM Mo）、N-乙酰-L-半胱氨酸（NAC）（100 μM）、Z-YVAD-氟甲基酮（YVAD）（10 μM）和 Mo 与 NAC 或 YVAD 的混合物处理鸭肾小管上皮细胞 12 h。通过 LDH 和 ELISA 试剂盒检测细胞上清液中 LDH 的释放和白细胞介素-1β（IL-1β）、白细胞介素-18（IL-18）的含量。通过流式细胞术用 MMP 和 ROS 试剂盒测定 MMP 和 ROS 水平。通过吖啶橙/溴化乙锭（AO/EB）复染法检测细胞的凋亡率。通过实时 qPCR 和 Western blot 分别检测细胞焦亡和凋亡相关因子的 mRNA 和蛋白水平。
过量 Mo 显著增加 LDH、IL-18 和 IL-1β的释放，并诱导 ROS 过度产生，增加细胞焦亡相关因子的 mRNA 和蛋白水平。NAC 和 YVAD 显著降低了 Mo 诱导的细胞焦亡。同时，YVAD 显著改变了凋亡相关因子的 mRNA 和蛋白水平，降低了细胞凋亡率。
过量 Mo 暴露可通过 ROS/NLRP3/Caspase-1 途径诱导鸭肾小管上皮细胞发生细胞焦亡，抑制 Caspase-1 依赖性细胞焦亡可能会减弱过量 Mo 诱导的凋亡。该研究为水禽过量 Mo 暴露肾毒性研究提供了理论依据，同时细胞焦亡与凋亡之间的相互作用为 Mo 诱导肾毒性的机制提供了新的视角。
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