摘要
目的探索重金属复合暴露对人神经母细胞瘤(SK-N-SH)细胞的毒性效应。方法将SK-N-SH细胞分别暴露于空白培养基(对照)和氯化锰(125、250、500、750、1000、1500、2000、4000、6000μmol/L),乙酸铅(500、750、1000、1500、2000、2500、3000μmol/L),氯化镉(5、10、15、20、30、40、60、80μmol/L)单一及二元、三元复合溶液24、48、96 h后,检测细胞存活率,计算锰、铅、镉的药物抑制浓度(IC)。其中,二元、三元复合溶液的浓度均为相应金属IC10、IC20、IC30、IC50的等毒效应比混合,分别用毒性单位法、混合毒性指数法对毒性效应进行评价。结果与对照组相比,不同浓度锰、铅、镉单独染毒不同时间SK-N-SH细胞的存活率均较低,除500μmol/L铅染毒24、96 h组和5μmol/L镉染毒24、48 h组外,差异均有统计学意义(P<0.05);随着锰、铅、镉单独染毒浓度的升高和染毒时间的延长,SK-N-SH细胞的存活率均呈下降趋势。三种金属对SK-N-SH的细胞急性毒性依次为镉(IC50:38.728μmol/L)>锰(IC50:442.739μmol/L)>铅(IC50:1624.850μmol/L)。同一金属不同染毒时间的IC50排序均为96 h<48 h<24 h。锰和镉复合暴露对SK-N-SH细胞的联合毒性在24 h均表现为拮抗作用,在48 h均表现为协同作用;96 h在毒效应为IC10、IC20时均表现为部分相加作用,在IC30、IC50时均表现为协同作用。镉和铅对SK-N-SH细胞的联合毒性在24 h毒效应为IC10、IC20、IC30时均表现为拮抗作用,在IC50时表现为部分相加作用;在48 h毒效应为IC10时表现为拮抗作用,在IC20、IC30、IC50时均表现为部分相加作用;在96 h时均表现为部分相加作用。铅和锰对SK-N-SH细胞的联合毒性在24、96 h毒效应为IC50时分别表现为简单相加、协同作用,其余均表现为部分相加作用。锰、铅、镉对SK-N-SH细胞的联合作用在24、96 h时均表现为部分相加作用;48 h时在IC10、IC20、IC30均表现为协同作用,在IC50时表现为部分相加作用。结论三种金属对SK-N-SH细胞的毒性均表现为随暴露时间的延长而增强的趋势。金属复合暴露呈现复杂的毒性效应,毒性效应与特定的金属组合、暴露时间、暴露浓度密切相关。
Objective To explore the toxicity of joint exposure to heavy metals on human neuroblastoma cells(SK-N-SH).Methods SK-N-SH cells were respectively exposed to blank medium(control),manganese chloride(125,250,500,750,1000,1500,2000,4000 and 6000μmol/L),lead acetate(500,750,1000,1500,2000,2500 and 3000μmol/L)and cadmium chloride(5,10,15,20,30,40,60 and 80μmol/L),with alone,binary and ternary metal mixtures for 24,48 and 96 h,then the inhibitory concentrations(IC)were calculated according to cell viability.The concentrations of binary and ternary composite solutions were set based on the levels of IC10,IC20,IC30 and IC50 for the corresponding metals,toxicity patterns of metal mixtures were evaluated by the toxicity unit and the mixed toxicity index.Results Compared with the control group,the cell viability of SK-N-SH cells alone exposed to three metals with different duration significantly decreased(P<0.05),except for 500μmol/L lead for 24 and 96 h,5μmol/L cadmium for 24 and 48 h;The cell viability decreased with the increase of exposure time and dose.The acute toxicity baesd on half inhibitory concentration(IC50)of the three metals ranked as cadmium(IC50:38.728μmol/L)>manganese(IC50:442.739μmol/L)>lead(IC50:1624.850μmol/L).The orders of IC50 ranked as 96 h<48 h<24 h for the same metal.The toxicity manner of manganese and cadmium co-exposure showed as antagonism effect in 24 h and synergy effect in 48 h;In 96 h,partial additive effect was showed in IC10,IC20 and synergy effect in IC30,IC50.In 24 h,the joint toxicity of cadmium and lead on SK-N-SH cells showed antagonistic effect in IC10,IC20,IC30 and partial additive effect in IC50;In 48 h,the toxic effect showed antagonistic effect in IC10 and partial additive effect in IC20,IC30,IC50,the toxic effect all showed partial additive effect in 96 h.The joint toxicity of lead and manganese on SK-N-SH cells showed simple additive and synergistic effect in IC50 in 24 and 96 h respectively,and partial additive effect in the rest treatments.Partially additive effect was observed in ternary metals(cadmium,lead and manganese)exposure for 24 and 96 h;In 48 h,the toxic effect showed synergistic effect in IC10,IC20,IC30 and partially additive effect in IC50.Conclusion Cytotoxicity of the three individual metals gets serious with time extension.Complex toxic patterns are presented in metal mixtures exposure,which are contributed to metal species,concentration and their exposure time.
作者
徐怡璐
吴彬
石倩倩
韦兰成
敖青青
孟玲
韦雪静
李习艺
唐深
王新航
陆彩玲
XU Yi-lu;WU Bin;SHI Qian-qian;WEI Lan-cheng;AO Qing-qing;MENG Ling;WEI Xue-jing;LI Xi-yi;TANG Shen;WANG Xin-hang;LU Cai-ling(School of Public Health,Guangxi Medical University,Nanning,Guangxi 530021,China;不详)
出处
《环境与健康杂志》
CAS
北大核心
2019年第8期679-684,共6页
Journal of Environment and Health
基金
国家自然科学基金(81760576,21567006,81460506,81860585).
