摘要
目的 观察低浓度含Mn^(2+)化合物和自噬抑制药物氯喹长时间共同作用对神经细胞株PC12细胞的毒性作用及机制。方法 取对数生长期PC12细胞,分别给予0(对照)、5、10、25、50、100、200μM氯化锰,5、10、25、50、100μM氯喹;染毒24 h。采用噻唑蓝(MTT)法检测氯化锰和氯喹对细胞存活率的影响。并联合给药,观察两种化合物共同作用24、48、72 h对细胞存活率的影响。进一步检测线粒体呼吸功能,探讨氯化锰和氯喹可能的毒性作用机理。结果 与空白对照组比较,氯化锰和氯喹单独给药均对细胞增殖活力具有抑制作用,其效应表现为给药剂量不断增加,而细胞存活率逐渐降低。实验选取对细胞增殖活力无显著影响的氯化锰和氯喹浓度分别为40μM和2.5μM;在此浓度下,与空白对照组相比,氯喹单独给药24、48和72 h,其细胞存活率和线粒体呼吸功能没有明显改变;氯化锰单独给药作用72 h,其线粒体呼吸功能未受影响;但氯化锰和氯喹联合给药共同作用72 h后,细胞的存活率和线粒体呼吸功能均显著降低(P<0.05)。结论 低浓度的氯化锰和氯喹长时间共同作用对PC12细胞具有明显毒性,其毒性机制可能与其对线粒体功能损伤有关。
Objective To observe the toxicity of low concentration Mn~(2+) compound combined with autophagy inhibitor chloroquine on nerve cell line PC12 cells for long-term and its mechanism.Methods PC12 cells at logarithmic growth stage were treated with 0(control),5,10,25,50,100,and 200 μM manganous chloride,and 5,10,25,50,and 100 μM chloroquine for 24 h,respectively.The effect of manganous chloride and chloroquine on cell viability was detected by MTT assay.The combined effect of the two compounds on cell viability was determined at 24,48 and 72 h,respectively.The mitochondrial respiratory function was further examined to explore the possible toxicity mechanism of manganous chloride and chloroquine.Results Compared with the control group,manganous chloride and chloroquine alone had inhibitory effect on cells survival in a concentration-dependent manner.Manganous chloride and chloroquine at concentrations of 40 μM and 2.5 μM,respectively,had no significant effect on cell survival.Compared with the control group,administration of 2.5 μM chloroquine alone for 24,48 and 72 h did not significantly change cell survival and mitochondrial respiratory function.Treatment of cells with manganous chloride alone at the concentration of 40 μM for 72 h did affect mitochondrial respiratory function.However,the cell survival and mitochondrial respiratory function in the combined administration of manganous chloride and chloroquine for 72 h were significant decreased(P<0.05).Conclusion The long-term combination of low-concentration manganous chloride and chloroquine produced an additive cytotoxicity on PC12 cells,and the toxicity mechanism may be related to the damage of mitochondrial function.
作者
付小龙
余子敬
杨永刚
FU Xiaolong;YU Zijing;YANG Yonggang(Key Laboratory of Basic Pharmacology of Ministry of Education and Joint International Research Laboratory of Ethnomedicine of Ministry of Education,Zunyi Medical University,Zunyi,Guizhou 563006,China;Taihe Hospital,Affiliated Hospital of Hubei University of Medicine,Shiyan,Hubei 442000,China)
出处
《公共卫生与预防医学》
2024年第4期16-19,共4页
Journal of Public Health and Preventive Medicine
基金
贵州省科技厅项目(黔科合基础-ZK[2024]一般274)
遵义市科技局联合基金项目(遵市科合HZ字[2022]422号)。
关键词
氯化锰
氯喹
环境和食品暴露
神经毒性
线粒体功能
MnCl2
Chloroquine
Environmental and food exposure
Neurotoxicity
Mitochondrial function
