基于神经血管单元的硫酸锰体外神经毒性研究  

Neurotoxicity of manganese sulfate in vitro based on neurovascular unit model

作　　者：王翀[1] 赵康峰[1] 顾雯[1] 张宏伟[1] WANG Chong;ZHAO Kang-feng;GU Wen;ZHANG Hong-wei(National Institute of Environmental Health,Chinese Center for Disease Control and Prevention,Beijing 100021,China)

机构地区：[1]中国疾病预防控制中心环境与健康相关产品安全所

出　　处：《环境与健康杂志》2019年第1期1-4,共4页Journal of Environment and Health

基　　金：国家自然科学基金(81372949)

摘　　要：目的探讨硫酸锰对三种离体培养脑细胞及两种细胞模型的毒性损伤。方法体外分离培养神经胶质细胞、脑微血管内皮细胞(BMECs)及海马神经元并建立神经血管单元模型(NVU)和单层BMECs模型。分别以125、250、500、1000、2000和4000μmol/L硫酸锰进行细胞毒性试验,观察125、250、500和1000μmol/L硫酸锰对细胞内钙离子浓度影响。结果与对照组比较,离体培养的三种脑细胞的细胞活性随染毒剂量的升高而降低,差异具有统计学意义(P<0.05);两种细胞模型海马神经元细胞活性无明显变化(P>0.05)。随染毒剂量的升高,海马神经元从500μmol/L开始,神经胶质细胞和BMECs从125μmol/L开始细胞内Ca^2+浓度高于对照组,差异均具有统计学意义(P<0.05);250和1000μmol/L硫酸锰染毒组BMECs模型海马神经元Ca^2+浓度高于对照组(P<0.05),而NVU模型无明显变化。结论硫酸锰对本次体外培养的三种脑细胞具有损伤作用。锰能够激活钙离子流,可能进而激活相关的细胞转导通路。BMECs与NVU的屏障作用可以降低锰对海马神经元的细胞毒性作用,NVU的屏障作用更显著。Objective To determine the neurotoxicity of manganese sulfate to three kinds of cultured neurons and two neural models.Methods Neuroglial cells,brain microvascular endothelial cells (BMECs) and hippocampal neurons were isolated and cultured in vitro,and a neurovascular unit model (NVU) and a single-layer BMECs model were established respectively.The cytotoxicities of manganese sulfate with concentrations of 125,250,500,1 000,2 000 and 4 000 μmol/L to three kinds of brain cells and two kinds of brain cell models cultured in vitro,and the concentrations of intracellular calcium in two kinds of brain cell models cultured in vitro exposed to manganese sulfate with concentrations of 125,250,500 and 1 000 μmol/L determined respectively.Results Compared with the control group,the cell viability of the three kinds of brain cells cultured in vitro significantly decreased with the increase of exposure dose (P<0.05),the cell viability of hippocampal neurons in two brain cell models exposed to manganese sulfate revealed no significantly differences compared with the control group (P>0.05).Compared with the control group,the concentration of intracellular Ca^2+ increased with the increases of manganese sulfate concentrations, and revealed significant differences in hippocampal neurons exposed to manganese sulfate with concentration higher than 500 μmol/L,as well as in the neuroglial cells and BMECs exposed to manganese sulfate with concentration higher than 125 μmol/L (P<0.05).The concentration of intracellular Ca^2+ significantly increased in hippocampal neurons of BMECs model cultured in vitro exposed to 250 μmol/L and 1 000 μmol/L manganese sulfate (P<0.05),and revealed no significant differences in hippocampal neurons of the NVU model exposed to manganese sulfate.Conclusion Manganese sulfate may damage the three kinds of brain cells cultured in vitro.Manganese may activate calcium current and the related cell transduction pathways. The toxicity of manganese to hippocampal neurons can be inhibited by the cell barrier

关 键 词：硫酸锰 体外神经毒性 神经胶质细胞 脑微血管内皮细胞(BMECs) 海马神经元 神经血管单元(NVU) 细胞内CA^2+ 

分 类 号：R994.6[医药卫生—毒理学]