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硫酸锌对酵母细胞的毒性作用
吴丽华1, 陈燕飞1, 仪慧兰,等2
1.太原师范学院 生物系;2.山西大学 生命科学学院
摘要:
【目的】研究硫酸锌对酵母细胞的毒性作用,为揭示高浓度锌的毒性作用机理提供依据。【方法】以模式生物酵母细胞为材料,在YPD培养基中添加终浓度为0,0.5,1,3,5和10 mmol/L的硫酸锌进行毒性试验。缓解组为抗氧化剂AsA、NO清除剂c-PTIO和Ca2+螯合剂EGTA分别与0.5或5 mmol/L硫酸锌同时作用。培养一定时间后,采用分光光度法、美蓝染色法和流式细胞术,研究硫酸锌对酵母细胞生长、细胞死亡率及胞内ROS、NO和Ca2+水平的影响。【结果】低浓度硫酸锌(0.5 mmol/L)促进酵母细胞生长,而高浓度硫酸锌(1~10 mmol/L)抑制酵母细胞生长,诱导细胞死亡,且细胞死亡率随着硫酸锌浓度升高和胁迫时间延长而逐渐升高。酵母细胞经5 mmol/L硫酸锌胁迫12或24 h后,胞内ROS、NO和Ca2+水平均显著高于对照组;但加入对应抗氧化剂AsA、NO清除剂c-PTIO和Ca2+螯合剂EGTA后,酵母细胞死亡率均显著低于硫酸锌单独处理组。【结论】高浓度硫酸锌可诱导酵母细胞死亡,在此过程中ROS、NO和Ca2+水平显著提高,从而造成毒性。
关键词:  硫酸锌  酵母细胞  毒性机理  细胞死亡
DOI:
分类号:
基金项目:国家自然科学基金项目(21307087);山西省应用基础研究计划青年科技研究基金项目(201601D021104);太原师范学院大学生创新项目(CXCY1971)
Cytotoxicity of zinc sulfate on Saccharomyces cerevisiae
WU Lihua,CHEN Yanfei,YI Huilan,et al
Abstract:
【Objective】In order to investigate the cellular mechanisms involved in high concentrations zinc-caused cytotocixity,the toxic effects of zinc sulfate on Saccharomyces cerevisiae was studied.【Method】Yeast cells were incubated in fresh yeast extract peptone dextrose (YPD) media containing zinc sulfate at concentrations of 0,0.5,1,3,5 and 10 mmol/L.In combination treatments,selected antagonists including ascorbic acid (AsA),nitric oxide (NO) scavenger 2-(4-carboxyphenyl)-4,4,5,5-teramethylimidazoline-1-oxyl-3-oxide (c-PTIO) and calcium chelator ethylene glycolbis-(2-aminoethyl)-tetraacetic acid (EGTA) were added into YPD media in the presence of 0.5 or 5 mmol/L zinc sulfate.After certain time,yeast cell growth was measured using a spectrophotometer,the cell death rate was determined by methylene blue staining,and the levels of intracellular reactive oxygen species (ROS),nitric oxide (NO) and calcium (Ca2+) were analyzed using a fluorescence activated cell sorter (FACS) Calibur at low flow rate.【Result】Zinc at low concentration of 0.5 mmol/L promoted cell growth,while zinc at high concentrations (1-10 mmol/L) inhibited cell growth and induced cell death.The cell mortality increased with the increase of zinc concentration and extension of stress time.The levels of intracellular ROS,NO and Ca2+ increased significantly in the yeast cells exposed to 5 mmol/L zinc for 12 or 24 h compared with the control.Antagonists AsA,c-PTIO,and EGTA significantly alleviated yeast cell death induced by zinc sulfate.【Conclusion】Cell death induced by high concentrations of zinc sulfate was associated with the increase of intercellular ROS,NO and Ca2+ levels.
Key words:  zinc sulfate  Saccharomyces cerevisiae cell  cytotoxicity mechanism  cell death

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