HOG1 MAPK参与调控亚砷酸钠诱导的酵母细胞凋亡
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摘要
【目的】本研究探讨了HOG1 MAPK在亚砷酸钠诱导酵母细胞凋亡中的作用。【方法】以酵母野生株BY4741及其HOG1突变株(ΔHOG1)为材料,研究了亚砷酸钠对酵母细胞生长、相对存活率和氧化损伤的影响,并采用流式细胞术检测了亚砷酸钠胁迫下酵母细胞凋亡率、ROS水平和线粒体膜电位的变化。【结果】亚砷酸钠可抑制酵母细胞生长,诱导细胞凋亡。在相同处理组中,ΔHOG1对亚砷酸钠更为敏感,表现为细胞存活率降低,凋亡率升高。在亚砷酸钠胁迫过程中,ΔHOG1胞内ROS水平和MDA含量显著高于野生株BY4741,而线粒体膜电位显著低于野生株。【结论】HOG1 MAPK可能通过影响胞内ROS水平和线粒体膜电位的变化调控亚砷酸钠诱导的酵母细胞凋亡。
[Objective] In this study, we studied the role of HOG1 MAPK in sodium arsenite-induced apoptosis in yeast cells. [Methods] Yeast wild-type(BY4741) and HOG1 mutant(ΔHOG1) strains were used to study the effects of sodium arsenite on the growth and relative survival rate and oxidative damages of yeast cells. Further, the apoptotic rate, intracellular reactive oxygen species(ROS) level, and mitochondrial membrane potential of the yeast cells under sodium arsenite-induced stress were determined by flow cytometry. [Results] Sodium arsenite inhibited the growth of yeast cells and induced their apoptosis. Compared to the wild-type strain in the same treatment group, ΔHOG1 strain showed higher sensitivity to sodium arsenite with a lower cell survival rate and higher apoptotic rate. Under sodium arsenite-induced stress, ΔHOG1 strain showed significantly higher intracellular ROS and malondialdehyde(MDA) levels than the wild-type BY4741 strain. On the contrary, the mitochondrial membrane potential of ΔHOG1 strain was significantly lower than that of the wild-type BY4741 strain.[Conclusion] These results indicated that HOG1 MAPK gene was involved in the regulation of sodium arsenite-induced apoptosis by affecting intracellular ROS level and changing ?ψM in yeast cells.
[Objective] In this study, we studied the role of HOG1 MAPK in sodium arsenite-induced apoptosis in yeast cells. [Methods] Yeast wild-type(BY4741) and HOG1 mutant(ΔHOG1) strains were used to study the effects of sodium arsenite on the growth and relative survival rate and oxidative damages of yeast cells. Further, the apoptotic rate, intracellular reactive oxygen species(ROS) level, and mitochondrial membrane potential of the yeast cells under sodium arsenite-induced stress were determined by flow cytometry. [Results] Sodium arsenite inhibited the growth of yeast cells and induced their apoptosis. Compared to the wild-type strain in the same treatment group, ΔHOG1 strain showed higher sensitivity to sodium arsenite with a lower cell survival rate and higher apoptotic rate. Under sodium arsenite-induced stress, ΔHOG1 strain showed significantly higher intracellular ROS and malondialdehyde(MDA) levels than the wild-type BY4741 strain. On the contrary, the mitochondrial membrane potential of ΔHOG1 strain was significantly lower than that of the wild-type BY4741 strain.[Conclusion] These results indicated that HOG1 MAPK gene was involved in the regulation of sodium arsenite-induced apoptosis by affecting intracellular ROS level and changing ?ψM in yeast cells.
引文
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[4]Azizur Rahman M,Hasegawa H,Mahfuzur Rahman M,Mazid Miah MA,Tasmin A.Arsenic accumulation in rice(Oryza sativa L.):human exposure through food chain.Ecotoxicology and Environmental Safety,2008,69(2):317-324.
[5]Chayapong J,Madhyastha H,Madhyastha R,Nurrahmah QI,Nakajima Y,Choijookhuu N,Hishikawa Y,Maruyama M.Arsenic trioxide induces ROS activity and DNA damage,leading to G0/G1 extension in skin fibroblasts through the ATM-ATR-associated Chk pathway.Environmental Science and Pollution Research,2017,24(6):5316-5325.
[6]Jomova K,Jenisova Z,Feszterova M,Baros S,Liska J,Hudecova D,Rhodes CJ,Valko M.Arsenic:toxicity,oxidative stress and human disease.Journal of Applied Toxicology,2011,31(2):95-107.
[7]Guo N,Peng ZL.MG132,a proteasome inhibitor,induces apoptosis in tumor cells.Asia-Pacific Journal of Clinical Oncology,2013,9(1):6-11.
[8]Kang YH,Lee SJ.The role of p38 MAPK and JNK in arsenic trioxide-induced mitochondrial cell death in human cervical cancer cells.Journal of Cellular Physiology,2008,217(1):23-33.
[9]O’Rourke SM,Herskowitz I,O’Shea EK.Yeast go the whole HOG for the hyperosmotic response.Trends in Genetics,2002,18(8):405-412.
[10]Azad GK,Singh V,Thakare MJ,Baranwal S,Tomar RS.Mitogen-activated protein kinase Hog1 is activated in response to curcumin exposure in the budding yeast Saccharomyces cerevisiae.BMC Microbiology,2014,14:317.
[11]Winkler A,Arkind C,Mattison CP,Burkholder A,Knoche K,Ota I.Heat stress activates the yeast high-osmolarity glycerol mitogen-activated protein kinase pathway,and protein tyrosine phosphatases are essential under heat stress.Eukaryotic Cell,2002,1(2):163-173.
[12]Panadero J,Pallotti C,Rodríguez-Vargas S,Randez-Gil F,Prieto JA.A downshift in temperature activates the high osmolarity glycerol(HOG)pathway,which determines freeze tolerance in Saccharomyces cerevisiae.Journal of Biological Chemistry,2006,281(8):4638-4645.
[13]Lawrence CL,Botting CH,Antrobus R,Coote PJ.Evidence of a new role for the high-osmolarity glycerol mitogen-activated protein kinase pathway in yeast:regulating adaptation to citric acid stress.Molecular and Cellular Biology,2004,24(8):3307-3323.
[14]Thorsen M,Di YJ,T?ngemo C,Morillas M,Ahmadpour D,Van der Does C,Wagner A,Johansson E,Boman J,Posas F,Wysocki R,Tamás MJ.The MAPK Hog1p modulates Fps1p-dependent arsenite uptake and tolerance in yeast.Molecular Biology of the Cell,2006,17(10):4400-4410.
[15]Wu LH,Yi HL,Zhang HF.Involvement of NO in sodium arsenite-induced yeast cell death.Acta Scientiae Circumstantiae,2012,32(10):2612-2616.(in Chinese)吴丽华,仪慧兰,张虎芳.NO参与亚砷酸钠诱导酵母细胞死亡的调控.环境科学学报,2012,32(10):2612-2616.
[16]Xiong B,Zhang LL,Xu HH,Yang Y,Jiang LH.Cadmium induces the activation of cell wall integrity pathway in budding yeast.Chemico-Biological Interactions,2015,240:316-323.
[17]Wu LH,Chen YF,Chen P,Yi HL.Sodium arsenite exposure affects the levels of antioxidant enzymes and lipid peroxidation in Saccharomyces cerevisiae.Asian Journal of Ecotoxicology,2016,11(3):302-307.(in Chinese)吴丽华,陈燕飞,陈鹏,仪慧兰.亚砷酸钠对酵母细胞抗氧化酶活性和脂质过氧化的影响.生态毒理学报,2016,11(3):302-307.
[18]潘军航.酿酒酵母耐铝机制中可能包含抗氧化作用.浙江大学硕士学位论文,2005.
[19]Campbell JS,Seger R,Graves JD,Graves LM,Jensen AM,Krebs EG.The MAP kinase cascade.Recent Progress in Hormone Research,1995,50:131-159.
[20]Bu SZ,Huang Q,Jiang YM,Min HB,Hou Y,Guo ZY,Wei JF,Wang JW,Ni X,Zheng SS.P38 mitogen-activated protein kinases is required for counteraction of 2-methoxyestradiol to estradiol-stimulated cell proliferation and induction of apoptosis in ovarian carcinoma cells via phosphorylation Bcl-2.Apoptosis,2006,11(3):413-425.
[21]Liao ZX,Wang F,Cao DX,Liu CL,Li YJ.Effect of p38MAPK pathway on isoflurane-induced neuronal apoptosis in hippocampus of neonatal rat.Chinese Pharmacological Bulletin,2014,30(12):1661-1666.(in Chinese)廖朝霞,王飞,曹德雄,柳垂亮,李玉娟.p38 MAPK信号通路在异氟醚诱导新生大鼠海马神经细胞凋亡的作用.中国药理学通报,2014,30(12):1661-1666.
[22]Liu AH,Gong XW,Wei J,Ming XY,Wang DA,Deng P,Luo SQ,Jiang Y.Effects of p38 MAPK gene knockout on arsenite-induced cell apoptosis.Chinese Journal of Pathophysiology,2008,24(5):892-895.(in Chinese)刘爱华,龚小卫,魏洁,明小燕,王达安,邓鹏,罗深秋,姜勇.p38 MAPK基因敲除对亚砷酸盐诱导细胞凋亡的影响.中国病理生理杂志,2008,24(5):892-895.
[23]Wu XC,Hu SJ,Qian KX.HOG-MAPK pathway in yeast.Chinese Journal of Cell Biology,2005,27(3):247-252.(in Chinese)吴雪昌,胡森杰,钱凯先.酵母HOG-MAPK途径.细胞生物学杂志,2005,27(3):247-252.
[24]Du L,Yu Y,Chen JS,Liu Y,Xia YJ,Chen Q,Liu XJ.Arsenic induces caspase-and mitochondria-mediated apoptosis in Saccharomyces cerevisiae.FEMS Yeast Research,2007,7(6):860-865.
[25]Zheng K,Pan JW,Ye L,Fu Y,Peng HZ,Wan BY,Gu Q,Bian HW,Han N,Wang JH,Kang B,Pan JH,Shao HH,Wang WZ,Zhu MY.Programmed cell death-involved aluminum toxicity in yeast alleviated by antiapoptotic members with decreased calcium signals.Plant Physiology,2007,143(1):38-49.
[26]Lee CH,Yu HS.Role of mitochondria,ROS,and DNAdamage in arsenic induced carcinogenesis.Frontiers in Bioscience,2016,8:312-320.
[27]Wang GY,Zhang T,Sun W,Wang HS,Yin F,Wang ZY,Zuo DQ,Sun MX,Zhou ZF,Lin BH,Xu J,Hua YQ,Li HQ,Cai ZD.Arsenic sulfide induces apoptosis and autophagy through the activation of ROS/JNK and suppression of Akt/m TORsignaling pathways in osteosarcoma.Free Radical Biology and Medicine,2017,106:24-37.
[28]Wu LH,Yi HL,Yi M.Assessment of arsenic toxicity using Allium/Vicia root tip micronucleus assays.Journal of Hazardous Materials,2010,176(1/3):952-956.
[29]Wu LH,Yi HL,Zhang HF.Reactive oxygen species and Ca2+are involved in sodium arsenite-induced cell killing in yeast cells.FEMS Microbiology Letters,2013,343(1):57-63.
[30]Chen YW,Yang CY,Huang CF,Hung DZ,Leung YM,Liu SH.Heavy metals,islet function and diabetes development.Islets,2009,1(3):169-176.
[31]Jomova K,Valko M.Advances in metal-induced oxidative stress and human disease.Toxicology,2011,283(2/3):65-87.
[32]Jomova K,Vondrakova D,Lawson M,Valko M.Metals,oxidative stress and neurodegenerative disorders.Molecular and Cellular Biochemistry,2010,345(1/2):91-104.
[33]Lu TH,Tseng TJ,Su CC,Tang FC,Yen CC,Liu YY,Yang CY,Wu CC,Chen KL,Huang DZ,Chen YW.Arsenic induces reactive oxygen species-caused neuronal cell apoptosis through JNK/ERK-mediated mitochondria-dependent and GRP 78/CHOP-regulated pathways.Toxicology Letters,2014,224(1):130-140.
[34]李雅楠.MAPK/线粒体凋亡途径在绵羊肺炎支原体致支气管上皮细胞氧化损伤中的作用机制研究.宁夏大学博士学位论文,2017.
[2]Liu SX,Athar M,Lippai I,Waldren C,Hei TK.Induction of oxyradicals by arsenic:implication for mechanism of genotoxicity.Proceedings of the National Academy of Sciences of the United States of America,2001,98(4):1643-1648.
[3]Waxman S,Anderson KC.History of the development of arsenic derivatives in cancer therapy.The Oncologist,2001,6(S2):3-10.
[4]Azizur Rahman M,Hasegawa H,Mahfuzur Rahman M,Mazid Miah MA,Tasmin A.Arsenic accumulation in rice(Oryza sativa L.):human exposure through food chain.Ecotoxicology and Environmental Safety,2008,69(2):317-324.
[5]Chayapong J,Madhyastha H,Madhyastha R,Nurrahmah QI,Nakajima Y,Choijookhuu N,Hishikawa Y,Maruyama M.Arsenic trioxide induces ROS activity and DNA damage,leading to G0/G1 extension in skin fibroblasts through the ATM-ATR-associated Chk pathway.Environmental Science and Pollution Research,2017,24(6):5316-5325.
[6]Jomova K,Jenisova Z,Feszterova M,Baros S,Liska J,Hudecova D,Rhodes CJ,Valko M.Arsenic:toxicity,oxidative stress and human disease.Journal of Applied Toxicology,2011,31(2):95-107.
[7]Guo N,Peng ZL.MG132,a proteasome inhibitor,induces apoptosis in tumor cells.Asia-Pacific Journal of Clinical Oncology,2013,9(1):6-11.
[8]Kang YH,Lee SJ.The role of p38 MAPK and JNK in arsenic trioxide-induced mitochondrial cell death in human cervical cancer cells.Journal of Cellular Physiology,2008,217(1):23-33.
[9]O’Rourke SM,Herskowitz I,O’Shea EK.Yeast go the whole HOG for the hyperosmotic response.Trends in Genetics,2002,18(8):405-412.
[10]Azad GK,Singh V,Thakare MJ,Baranwal S,Tomar RS.Mitogen-activated protein kinase Hog1 is activated in response to curcumin exposure in the budding yeast Saccharomyces cerevisiae.BMC Microbiology,2014,14:317.
[11]Winkler A,Arkind C,Mattison CP,Burkholder A,Knoche K,Ota I.Heat stress activates the yeast high-osmolarity glycerol mitogen-activated protein kinase pathway,and protein tyrosine phosphatases are essential under heat stress.Eukaryotic Cell,2002,1(2):163-173.
[12]Panadero J,Pallotti C,Rodríguez-Vargas S,Randez-Gil F,Prieto JA.A downshift in temperature activates the high osmolarity glycerol(HOG)pathway,which determines freeze tolerance in Saccharomyces cerevisiae.Journal of Biological Chemistry,2006,281(8):4638-4645.
[13]Lawrence CL,Botting CH,Antrobus R,Coote PJ.Evidence of a new role for the high-osmolarity glycerol mitogen-activated protein kinase pathway in yeast:regulating adaptation to citric acid stress.Molecular and Cellular Biology,2004,24(8):3307-3323.
[14]Thorsen M,Di YJ,T?ngemo C,Morillas M,Ahmadpour D,Van der Does C,Wagner A,Johansson E,Boman J,Posas F,Wysocki R,Tamás MJ.The MAPK Hog1p modulates Fps1p-dependent arsenite uptake and tolerance in yeast.Molecular Biology of the Cell,2006,17(10):4400-4410.
[15]Wu LH,Yi HL,Zhang HF.Involvement of NO in sodium arsenite-induced yeast cell death.Acta Scientiae Circumstantiae,2012,32(10):2612-2616.(in Chinese)吴丽华,仪慧兰,张虎芳.NO参与亚砷酸钠诱导酵母细胞死亡的调控.环境科学学报,2012,32(10):2612-2616.
[16]Xiong B,Zhang LL,Xu HH,Yang Y,Jiang LH.Cadmium induces the activation of cell wall integrity pathway in budding yeast.Chemico-Biological Interactions,2015,240:316-323.
[17]Wu LH,Chen YF,Chen P,Yi HL.Sodium arsenite exposure affects the levels of antioxidant enzymes and lipid peroxidation in Saccharomyces cerevisiae.Asian Journal of Ecotoxicology,2016,11(3):302-307.(in Chinese)吴丽华,陈燕飞,陈鹏,仪慧兰.亚砷酸钠对酵母细胞抗氧化酶活性和脂质过氧化的影响.生态毒理学报,2016,11(3):302-307.
[18]潘军航.酿酒酵母耐铝机制中可能包含抗氧化作用.浙江大学硕士学位论文,2005.
[19]Campbell JS,Seger R,Graves JD,Graves LM,Jensen AM,Krebs EG.The MAP kinase cascade.Recent Progress in Hormone Research,1995,50:131-159.
[20]Bu SZ,Huang Q,Jiang YM,Min HB,Hou Y,Guo ZY,Wei JF,Wang JW,Ni X,Zheng SS.P38 mitogen-activated protein kinases is required for counteraction of 2-methoxyestradiol to estradiol-stimulated cell proliferation and induction of apoptosis in ovarian carcinoma cells via phosphorylation Bcl-2.Apoptosis,2006,11(3):413-425.
[21]Liao ZX,Wang F,Cao DX,Liu CL,Li YJ.Effect of p38MAPK pathway on isoflurane-induced neuronal apoptosis in hippocampus of neonatal rat.Chinese Pharmacological Bulletin,2014,30(12):1661-1666.(in Chinese)廖朝霞,王飞,曹德雄,柳垂亮,李玉娟.p38 MAPK信号通路在异氟醚诱导新生大鼠海马神经细胞凋亡的作用.中国药理学通报,2014,30(12):1661-1666.
[22]Liu AH,Gong XW,Wei J,Ming XY,Wang DA,Deng P,Luo SQ,Jiang Y.Effects of p38 MAPK gene knockout on arsenite-induced cell apoptosis.Chinese Journal of Pathophysiology,2008,24(5):892-895.(in Chinese)刘爱华,龚小卫,魏洁,明小燕,王达安,邓鹏,罗深秋,姜勇.p38 MAPK基因敲除对亚砷酸盐诱导细胞凋亡的影响.中国病理生理杂志,2008,24(5):892-895.
[23]Wu XC,Hu SJ,Qian KX.HOG-MAPK pathway in yeast.Chinese Journal of Cell Biology,2005,27(3):247-252.(in Chinese)吴雪昌,胡森杰,钱凯先.酵母HOG-MAPK途径.细胞生物学杂志,2005,27(3):247-252.
[24]Du L,Yu Y,Chen JS,Liu Y,Xia YJ,Chen Q,Liu XJ.Arsenic induces caspase-and mitochondria-mediated apoptosis in Saccharomyces cerevisiae.FEMS Yeast Research,2007,7(6):860-865.
[25]Zheng K,Pan JW,Ye L,Fu Y,Peng HZ,Wan BY,Gu Q,Bian HW,Han N,Wang JH,Kang B,Pan JH,Shao HH,Wang WZ,Zhu MY.Programmed cell death-involved aluminum toxicity in yeast alleviated by antiapoptotic members with decreased calcium signals.Plant Physiology,2007,143(1):38-49.
[26]Lee CH,Yu HS.Role of mitochondria,ROS,and DNAdamage in arsenic induced carcinogenesis.Frontiers in Bioscience,2016,8:312-320.
[27]Wang GY,Zhang T,Sun W,Wang HS,Yin F,Wang ZY,Zuo DQ,Sun MX,Zhou ZF,Lin BH,Xu J,Hua YQ,Li HQ,Cai ZD.Arsenic sulfide induces apoptosis and autophagy through the activation of ROS/JNK and suppression of Akt/m TORsignaling pathways in osteosarcoma.Free Radical Biology and Medicine,2017,106:24-37.
[28]Wu LH,Yi HL,Yi M.Assessment of arsenic toxicity using Allium/Vicia root tip micronucleus assays.Journal of Hazardous Materials,2010,176(1/3):952-956.
[29]Wu LH,Yi HL,Zhang HF.Reactive oxygen species and Ca2+are involved in sodium arsenite-induced cell killing in yeast cells.FEMS Microbiology Letters,2013,343(1):57-63.
[30]Chen YW,Yang CY,Huang CF,Hung DZ,Leung YM,Liu SH.Heavy metals,islet function and diabetes development.Islets,2009,1(3):169-176.
[31]Jomova K,Valko M.Advances in metal-induced oxidative stress and human disease.Toxicology,2011,283(2/3):65-87.
[32]Jomova K,Vondrakova D,Lawson M,Valko M.Metals,oxidative stress and neurodegenerative disorders.Molecular and Cellular Biochemistry,2010,345(1/2):91-104.
[33]Lu TH,Tseng TJ,Su CC,Tang FC,Yen CC,Liu YY,Yang CY,Wu CC,Chen KL,Huang DZ,Chen YW.Arsenic induces reactive oxygen species-caused neuronal cell apoptosis through JNK/ERK-mediated mitochondria-dependent and GRP 78/CHOP-regulated pathways.Toxicology Letters,2014,224(1):130-140.
[34]李雅楠.MAPK/线粒体凋亡途径在绵羊肺炎支原体致支气管上皮细胞氧化损伤中的作用机制研究.宁夏大学博士学位论文,2017.