汞盐诱导烟草悬浮细胞产生细胞编程死亡

doi:10.3864/j.issn.0578-1752.2012.20.004

摘要/Abstract

摘要： 【目的】阐明氯化汞处理烟草悬浮细胞产生的细胞死亡具有细胞编程死亡（programmed cell death，PCD）的特征和分子机制。【方法】采用形态和生物化学方法观察氯化汞处理细胞后的变化，并采用药理学方法，通过添加抑制剂和抗氧化剂阐明氯化汞引发的信号转导途径。【结果】氯化汞诱导产生的细胞死亡具有PCD特征，包括染色质浓缩、细胞核的TUNEL检测呈阳性和DNA ladder的形成。氯化汞诱导烟草悬浮细胞过氧化氢积累，过氧化氢酶和抗坏血酸等抗氧化剂能降低细胞死亡率。蛋白激酶抑制剂、磷脂酶C和磷脂酶D的抑制剂能显著地降低氯化汞引起的细胞死亡和过氧化氢积累。【结论】氯化汞引起的细胞死亡是一种PCD，蛋白激酶和磷脂信号介导的过氧化氢积累参与细胞死亡过程。

关键词: 烟草, 氯化汞, 细胞编程死亡

Abstract: 【Objective】 The objective of this study is to investigate the characteristics of cell death and molecular mechanisms induced by HgCl2. 【Method】 HgCl2-induced cell death was studied in suspension-cultured tobacco cells via neutral red staining, a set of cellular morphological and biochemical characteristics was elucidated in the occurrence of cell death, and pharmacological method was used to explore the mechanisms activated by HgCl2. 【Result】 Within 24 and 48 h, HgCl2 treatment induced cell death in a concentration-dependent manner. Tobacco cells treated with HgCl2 showed typical features of PCD such as chromatin condensation, TUNEL-positive nuclei and DNA ladder formation. HgCl2-induced cell death was accompanied by an increasing production of H2O2. Consistently, the addition of the antioxidants greatly reduced cell death. Pretreatment with protein kinase inhibitor staurosporin, phospholipase C (PLC) inhibitors U73122 and neomycin, and phospholipase D (PLD) inhibitor 1-butanol significantly diminished HgCl2-induced cell death and H2O2 accumulation. 【Conclusion】 HgCl2 induces PCD exhibiting apoptotic-like features. The cell death process requires increased H2O2 production regulated by activation of protein phosphorylation and phospholipid signaling pathways.

Key words: tobacco, HgCl2, programmed cell death

柳金伟, 焦娇, 王蕾, 王莲莲, 褚小静, 梁元存. 汞盐诱导烟草悬浮细胞产生细胞编程死亡[J]. 中国农业科学, 2012, 45(20): 4149-4155.

LIU Jin-Wei, JIAO Jiao, WANG Lei, WANG Lian-Lian, CHU Xiao-Jing, LIANG Yuan-Cun. HgCl2-Induced Programmed Cell Death in Tobacco Cells[J]. Scientia Agricultura Sinica, 2012, 45(20): 4149-4155.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.chinaagrisci.com/CN/10.3864/j.issn.0578-1752.2012.20.004

https://www.chinaagrisci.com/CN/Y2012/V45/I20/4149

参考文献

[1]Zahir F, Rizwi S J, Haq S K, Khan R H. Low dose mercury toxicity and human health. Environmental Toxicology and Pharmacology, 2005, 20: 351-360.

[2]陆安详, 孙江, 王纪华, 董文光, 韩平, 张国光, 王开义, 潘立刚. 北京农田土壤重金属年际变化及其特征分析. 中国农业科学, 2011, 44(18): 3778-3789.

Lu A X, Sun J, Wang J H, Dong W G, Han P, Zhang G G, Wang K Y, Pan L G. Annual variability and characteristics analysis of heavy metals in agricultural soil of Beijing. Scientia Agricultura Sinica, 2011, 44(18): 3778-3789. (in Chinese)

[3]Siegel S M, Puerner N J, Speitel T W. Release of volatile mercury from vascular plants. Physiologia Plantarum, 1974, 32: 174-176.

[4]Bernier M, Popovic R, Carpentier R. Mercury inhibition at the donor side of photosystem II is reversed by chloride. FEBS Letters, 1993, 321(1): 19-23.

[5]Johansson I, Karlsson M, Johanson U, Larsson C, Kjellbom P. The role of aquaporins in cellular and whole plant water balance. Biochimica et Biophysica Acta, 2000, 1465: 324-342.

[6]Cho U H, Park J O. Mercury-induced oxidative stress in tomato seedlings. Plant Science, 2000, 156: 1-9.

[7]孙斌, 李多川, 慈小燕, 郭润芳, 王颖. 小麦叶片β-1,3-葡聚糖酶的诱导、纯化与抗菌活性. 植物生理与分子生物学报, 2004, 30(4): 399-404.

Sun B, Li D C, Ci X Y, Guo R F, Wang Y. Induction, purification and antifungal activity of β-1, 3-glucanase from wheat leaves. Journal of Plant Physiology and Molecular Biology, 2004, 30(4): 399-404. (in Chinese)

[8]Steller H. Mechanisms and genes of cellular suicide. Science, 1995, 267: 1445-1449.

[9]Jones A M. Programmed cell death in development and defense. Plant Physiology, 2001, 125: 94-97.

[10]Zong W X, Thompson C B. Necrotic death as a cell fate. Genes & Development, 2006, 20: 1-15.

[11]Overmyer K, Brosché M, Kangasjärvi J. Reactive oxygen species and hormonal control of cell death. Trends in Plant Science, 2003, 8: 335-342.

[12]Zawoznik M S, Groppa M D, Tomaro M L, Benavides M P. Endogenous salicylic acid potentiates cadmium-induced oxidative stress in Arabidopsis thaliana. Plant Science, 2007, 173: 190-197.

[13]Yakimova E T, Kapchina-Toteva V M, Woltering E J. Signal transduction events in aluminum-induced cell death in tomato suspension cells. Journal of Plant Physiology, 2007, 164: 702-708.

[14]Yakimova E T, Kapchina-Toteva V M, Laarhoven L J, Harren F M, Woltering E J. Involvement of ethylene and lipid signaling in cadmium-induced programmed cell death in tomato suspension cells. Plant Physiology and Biochemistry, 2006, 44: 581-589.

[15]De Michele R, Vurro E, Rigo C, Costa A, Elviri L, Di Valentin M, Careri M, Zottini M, Sanitá di Toppi L, Lo Schiavo F. Nitric oxide is involved in cadmium-induced programmed cell death in Arabidopsis suspension cultures. Plant Physiology, 2009, 150: 217-228.

[16]刘菲, 刘辉, 梁元存, 刘爱新, 魏芳芳, 林琎. 核黄素诱导烟草悬浮细胞酚类物质和木质素的积累. 中国农业科学，2009, 42(9): 3230-3235.

Liu F, Liu H, Liang Y C, Liu A X, Wei F F, Lin J. Induction of phenolic compounds and lignin in cultured tobacco cells treated with riboflavin. Scientia Agricultura Sinica, 2009, 42(9): 3230-3235. (in Chinese)

[17]Binet M N, Humbert C, Lecourieux D, Vantard M, Pugin A. Disruption of microtubular cytoskeleton induced by cryptogein, an elicitor of hypersensitive response in tobacco cells. Plant Physiology, 2001, 125: 564-572.

[18]Bellincampi D, Dipierro N, Salvi G, Cervone F, De Lorenzo G. Extracellular H2O2 induced by oligogalacturonides is not involved in the inhibition of the auxine-regulated rolB gene expression in tobacco leaf explants. Plant Physiology, 2000, 122: 1379-1385.

[19]Kressel M, Groscurth P. Distinction of apoptotic and necrotic cell death by in situ labelling of fragmented DNA. Cell and Tissue Research, 1994, 278: 549-556.

[20]Yun L J, Chen W L. SA and ROS are involved in methyl salicylate- induced programmed cell death in Arabidopsis thaliana. Plant Cell Reports, 2011, 30: 1231-1239.

[21]Sharma S S, Dietz K. The relationship between metal toxicity and cellular redox imbalance. Trends in Plant Science, 2008, 14: 1360-1385.

[22]Mittler R. Oxidative stress, antioxidants and stress tolerance. Trends in Plant Science, 2002, 7: 405-410.

[23]Johnson S M, Doherty S J, Croy R R D. Biphasic superoxide generation in potato tubers. A self amplifying response to stress. Plant Physiology, 2003, 131: 1440-1449.

[24]Iannone M F, Rosales E P, Groppa M D, Benavides M P. Reactive oxygen species formation and cell death in catalase-deficient tobacco leaf disks exposed to cadmium. Protoplasma, 2010, 245: 15-27.

[25]Terres M A, Dangl J L, Jones J D G. Arabidopsis gp91phox homologues AtrbohD and AtrbohF are required for accumulation of reactive oxygen intermediates in the plant defense response. Proceedings of the National Academy of Sciences of the United States of America, 2002, 99(1): 517-522.

[26]Chou T S, Chao Y Y, Kao C H. Involvement of hydrogen peroxide in heat shock-and cadmium-induced expression of ascorbate peroxidase and glutathione reductase in leaves of rice seedlings. Journal of Plant Physiology, 2012, 169: 478-486.

[27]Schützendübel A, Schwanz P, Teichmann T, Gross K, Langenfeld-Heyser R, Godbold D L, Polle A. Cadmium-induced changes in antioxidative systems, hydrogen peroxide content, and differentiation in Scots pine roots. Plant Physiology, 2001, 127: 887-898.

[28]Viard M P, Martin F, Pugin A, Ricci P, Blein J P. Protein phosphorylation is induced in tobacco cells by the elicitor cryptogein. Plant Physiology, 1994, 104: 1245-1249.