胞外ATP对AlCl3诱导的细胞死亡过程中胞内H2O2和Ca2+水平的影响及其生理机制分析
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引用本文:达梦婷,石珍珍,庞海龙,贾凌云,孙 坤,冯汉青.胞外ATP对AlCl3诱导的细胞死亡过程中胞内H2O2和Ca2+水平的影响及其生理机制分析[J].西北植物学报,2019,39(6):1033~1041
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达梦婷,石珍珍,庞海龙,贾凌云,孙 坤,冯汉青* (西北师范大学 生命科学学院兰州 730070) 
基金项目:国家自然科学基金(31870246, 31560070);
中文摘要:该实验以烟草悬浮细胞 BY 2 为材料,在烟草悬浮细胞中分别加入0.05、0.10、0.15、0.20 mmol·L-1AlCl3,以等体积去离子水处理的悬浮细胞液为对照,并依据前述实验结果选择0.15 mmol·L-1 AlCl3,分别添加5 mmol·L-1 DMTU(H2O2 抑制剂)、20 μmol·L-1CaCl2、15 μmol·L-1 LaCl3(Ca2+通道抑制剂)和50 μmol·L-1 ATP设计多项处理,分析胞外ATP(eATP)对铝离子(Al3+)胁迫引起的植物细胞死亡及其胞内H2O2、Ca2+的影响,以揭示Al3+胁迫下植物调节细胞死亡的可能机制,进一步扩展对eATP功能的认知。结果显示:(1)随着 AlCl3 胁迫浓度的提高,细胞死亡水平和胞内H2O2水平上升,而胞内Ca2+和eATP水平则逐渐降低。(2)外援施加H2O2抑制剂 DMTU(二甲基硫脲)和Ca2+能够有效缓解AlCl3诱导的细胞死亡水平的上升;而Ca2+通道抑制剂LaCl3(三氯化镧)则加剧了AlCl3胁迫下的细胞死亡。(3)在AlCl3胁迫下对细胞添加外源ATP,能够缓解AlCl3胁迫下胞内H2O2水平上升和Ca2+水平下降的同时,并显著降低AlCl3胁迫导致的细胞死亡。研究表明, Al3+以剂量依赖的模式提升细胞死亡和细胞内H2O2的水平并降低胞内Ca2+和eATP水平,AlCl3诱导的细胞死亡受到H2O2和Ca2+水平变化的调节,eATP可以通过调节H2O2与Ca2+水平缓解AlCl3诱导的细胞死亡。推测Al3+胁迫可能通过抑制钙离子通道而破坏了细胞内H2O2和Ca2+之间的协同关系,外源ATP对Al3+诱导H2O2上升的缓解作用可能是由于其提升了细胞的抗氧化能力。
中文关键词:AlCl3胁迫  细胞死亡  胞外ATP(eATP)  H2O2  Ca2+
 
Extracellular ATP Alleviates the Aluminium induced Cell Death by Regulating H2O2 and Ca2+ Levels
Abstract:In this experiment, tobacco suspension cell BY 2 was used as the material, and 0.05, 0.10, 0.15, 0.20 mmol·L-1 AlCl3 was added to the tobacco suspension cells, and the suspension cell solution treatment with equal volume of deionization water was used as a control. And, according to the above experimental results, 0.15 mmol·L-1 AlCl3 was selected, 5 mmol·L-1 DMTU (H2O2 inhibitor), 20 μmol·L-1 CaCl2, 15 μmol·L-1 LaCl3 (Ca2+ channel inhibitor) and 50 μmol·L-1 ATP was used to analyze the effects of extra cellular ATP (eATP) on plant cell death and intra cellular H2O2 and Ca2+ induced by aluminum ion (Al3+) stress and to reveal plant the possible mechanism of eATP in regulating cell death under Al3+ stress, and further extend the perception of eATP function. The results showed that: (1) with the increase of AlCl3 concentration, the levels of cell death and intracellular H2O2 were increased, while the levels of intracellular Ca2+and eATP were decreased; (2) Application of DMTU (dimethylthiourea, a scavenger of H2O2) and exogenous Ca2+ effectively attenuated the AlCl3 induced cell death, while barium trichloride (LaCl3, the Ca2+ channel inhibitor) aggravated the AlCl3 induced cell death, indicating that AlCl3 induced cell death is regulated by H2O2 and Ca2+; (3) The addition of exogenous ATP effectively impeded the increase of intracellular H2O2 production and decrease of intracellular Ca2+ under AlCl3 stress, and the cell death induced by AlCl3 were also alleviated by exogenous ATP. It can be seen that aluminum induced cell death is regulated by changes in H2O2 and Ca2+ levels, and extracellular ATP affects the aluminum induced cell death by regulating H2O2 and Ca2+ levels.Theses observations indicated that the level of cell death and intracellular H2O2 were increased, while the level of intracellular Ca2+ and eATP were decreased in a dose dependent manner under Al3+ stress. It can be seen that aluminum induced cell death is regulated by changes in H2O2 and Ca2+ levels, and extracellular ATP affects the aluminum induced cell death by regulating H2O2 and Ca2+ levels. We predicated that Al3+ stress may destroy the synergistic relationship between H2O2 and Ca2+ in cells by inhibiting calcium channel, and the alleviation effect of exogenous ATP on Al3+ induced H2O2 elevation may be due to its enhanced antioxidant capacity.
keywords:AlCl3 stress  cell death  extracellular ATP  H2O2  Ca2+
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