《植物生理学报》 2021, 57(4): 838-846

细胞外ATP参与调节抗霉素A诱导的植物细胞死亡

王建荷¹，王娟娟¹，庞海龙¹，贾凌云¹，孙坤¹，张继^1,2，冯汉青^1,*

¹西北师范大学生命科学学院, 兰州730070；²西北师范大学新农村发展研究院, 兰州730070

通信作者：冯汉青；E-mail: fenghanq@nwnu.edu.cn

摘　要：

选用烟草悬浮细胞(Nicotiana tabacum ‘Bright Yellow-2’)为试验材料, 研究细胞外ATP (extracellular ATP , eATP)对抗霉素A (antimycin A , AA)所诱导的细胞死亡的影响及可能的调节机制。结果表明, AA可引起细胞死亡和细胞活性氧(reactive oxygen species, ROS)水平的上升, 外源施加ROS清除剂DMTU (N,N′-dimethylthiourea)可缓解AA诱导的细胞死亡水平的上升。另外, AA导致细胞eATP水平的下降; 而用外源ATP缓解AA诱导的eATP下降的同时, AA诱导的细胞死亡和ROS水平的上升也被缓解。进一步试验发现, Ca²⁺对AA诱导的细胞死亡和ROS水平的影响与eATP的行为相似, 且AA处理下的Ca²⁺水平受到了eATP的调控; 更为重要的是, 质膜Ca²⁺通道抑制剂或Ca²⁺螯合剂均可消除eATP对AA诱导的细胞死亡上升的缓解作用。上述观察表明, eATP能够调节AA诱导的细胞死亡, 这种作用很可能联系着eATP对ROS的影响且依赖于Ca²⁺作为其下游信号。

关键词：细胞外ATP; 抗霉素A; 细胞死亡; ROS; Ca2+

收稿：2021-01-22   修定：2021-03-04

资助：国家自然科学基金(31870246和31560070)、甘肃省重点研发计划(18YF1NA051)、甘肃省引导科技创新发展专项资金(2019ZX- 05)、甘肃省高等学校科研项目(2015A-007)、甘肃省高校基本科研业务费和西北师范大学青年创新团队项目。

Extracellular ATP is involved in the regulation of antimycin A-induced plant cell death

WANG Jianhe¹, WANG Juanjuan¹, PANG Hailong¹, JIA Lingyun¹, SUN Kun¹, ZHANG Ji1^1,2, FENG Hanqing^1,*

¹College of Life Sciences, Northwest Normal University, Lanzhou 730070, China; ²New Rural Development Research Institute, Northwest Normal University, Lanzhou 730070, China

Corresponding author: FENG Hanqing; E-mail: fenghanq@nwnu.edu.cn

Abstract:

Tobacco suspension cells (Nicotiana tabacum ‘Bright Yellow-2’) were used as experimental materials to study the effects and possible adjustment mechanisms of extracellular ATP (eATP) on cell death induced by antimycin A (AA). The results showed that AA could cause an increase in cell death and reactive oxygen species (ROS) levels, and the application of ROS scavenger DMTU (N,N′-dimethylthiourea)
could effectively alleviate the increase of AA-induced cell death. In addition, AA induced a decrease in cell eATP levels. Exogenous ATP was used to alleviate AA-induced eATP decrease, meanwhile AA-induced cell death and increase in ROS levels were also alleviated. Further experiments found that the effect of Ca²⁺ on AA-induced cell death and ROS levels was similar to the behavior of eATP, and eATP could regulate the Ca²⁺ levels under AA treatment. More importantly, plasma membrane Ca²⁺ channel inhibitor or Ca²⁺ chelator could eliminate the alleviating effect of eATP on the increase in cell death induced by AA. The above observations indicate that eATP can regulate AA-induced cell death, and this regulation is probably related to the effect of eATP on ROS and depends on Ca2+ as its downstream signal.

Key words: extracellular ATP; antimycin A; cell death; ROS; Ca2+