Department of Toxicology, School of Public Health, Guangxi Medical University, 22 Shuang-Yong Rd., Nanning, 530021, Guangxi, China.
Key Laboratory of Prevention and Control of Highly Prevalent Diseases in Guangxi Colleges and Universities, Medical University of Guangxi, Nanning, 530021, China.
Biol Trace Elem Res. 2024 May;202(5):2241-2252. doi: 10.1007/s12011-023-03792-z. Epub 2023 Jul 27.
The aim of study was to address the effects of manganese and iron, alone and in combination, on apoptosis of BV2 microglia cells, and to determine if combined exposure to these metals augments their individual toxicity. We used a murine microglial BV2 cell line. Cell cytotoxicity was analyzed by propidium iodide (PI) exclusion assay. Cell ROS production was analyzed by 2', 7'-dichlorofluorescin diacetate (DCFH-DA) probe staining. Pro-inflammatory cytokine production was monitored by ELISA. Cell apoptosis was analyzed by PE Annexin V/7-AAD staining. Mitochondrial membrane integrity was analyzed by flow cytometry. We used immunoblotting to analyze the effect of manganese, iron alone, or their combined exposure on the activation of caspase9, P53, Bax, and Bcl2 apoptosis signaling pathways. Caspase3 activity was determined using a Colorimetric. Manganese, iron, and their combined exposure for 24 h induced the activation of BV2 microglia cells and increased ROS production and the expression of the inflammatory cytokines, IL-1β and TNF-α. And we also found that the apoptosis rate increased, mitochondrial membrane potential decreased, apoptosis-related proteins caspase9, P53, Bax, and Bcl2 expression increased, and caspase3 activity increased. Furthermore, we found that combined manganese-iron cytotoxicity was lower than that induced by manganese exposure alone. Manganese, iron alone, or their combination exposure can induce apoptosis in glial cells. Iron can reduce the toxicity of manganese, and there is an antagonistic effect between manganese and iron.
本研究旨在探讨锰和铁单独及联合作用对 BV2 小胶质细胞凋亡的影响，并确定这两种金属联合暴露是否会增强其各自的毒性。我们使用了一种鼠源小胶质细胞 BV2 细胞系。用碘化丙啶（PI）排除法分析细胞细胞毒性。通过 2'，7'-二氯荧光素二乙酸酯（DCFH-DA）探针染色分析细胞 ROS 产生。通过 ELISA 监测促炎细胞因子的产生。通过 PE Annexin V/7-AAD 染色分析细胞凋亡。通过流式细胞术分析线粒体膜完整性。我们用免疫印迹分析锰、铁单独或联合暴露对 caspase9、P53、Bax 和 Bcl2 凋亡信号通路激活的影响。用比色法测定 caspase3 活性。锰、铁及其联合暴露 24 h 可诱导 BV2 小胶质细胞激活，增加 ROS 产生和促炎细胞因子 IL-1β和 TNF-α的表达。我们还发现凋亡率增加，线粒体膜电位降低，凋亡相关蛋白 caspase9、P53、Bax 和 Bcl2 表达增加，caspase3 活性增加。此外，我们发现锰铁联合毒性低于单独锰暴露诱导的毒性。锰、铁单独或联合暴露均可诱导神经胶质细胞凋亡。铁可以降低锰的毒性，锰和铁之间存在拮抗作用。
