模拟微重力对金黄色葡萄球菌基因表达及表型的影响
|
摘要
目的研究金黄色葡萄球菌在低剪切力模拟微重力系统(low-shear modeled microgravity,LSMMG)条件下连续培养14d的表型变化及基因表达改变。方法利用旋转细胞培养系统模拟微重力环境对金黄色葡萄球菌连续传代培养14d,对菌株进行增殖速率、耐酸性和生物膜形成的测定,评估金黄色葡萄球菌的表型变化。利用转录组测序检测模拟微重力条件下差异表达的基因,与表型作比对。结果模拟微重力导致金黄色葡萄球菌增殖速率降低,耐酸性增强,生物膜形成能力增强,耐碱性下调;共有172个显著差异表达基因(P<0.05),其中20个上调,152个下调。36个营养代谢相关差异表达基因中32个表达下降,1个与耐碱相关基因表达下调。结论模拟微重力引起金黄色葡萄球菌表型及相应的基因表达变化,其中生物膜形成能力的增强可能对航天飞行造成潜在威胁。
Objective To detect the phenotypic changes and gene changes of Staphylococcus aureus under low-shear modeled microgravity(LSMMG)conditions.Methods Continuous culture of Staphylococcus aureus was performed using a rotating cell culture system to simulate the microgravity environment.The proliferation rate,acid resistance and biofilm formation of the strains were measured.The phenotypic changes of Staphylococcus aureus were evaluated.Transcriptome sequencing was used to detect the differentially expressed genes under simulated microgravity conditions and then they were compared with the phenotypes.Results Simulated microgravity resulted in a decrease in the proliferation rate of Staphylococcus aureus,an increase in the acid resistance,an enhancement in biofilm formation,and a down-regulation of alkalinity.There were a total of 172 differentially expressed genes in the simulated microgravity group(P<0.05,of which 20 were up-regulated and 152 were down-regulated),there were 36 differentially expressed genes related to nutrition metabolism,32 of which decreased in expression,and 1 was associated with decreased expression of alkali-resistant genes.Conclusion Simulated microgravity can cause the phenotype and corresponding genetic changes in Staphylococcus aureus,and the enhancement of biofilm formation may pose a potential threat to space flight.
Objective To detect the phenotypic changes and gene changes of Staphylococcus aureus under low-shear modeled microgravity(LSMMG)conditions.Methods Continuous culture of Staphylococcus aureus was performed using a rotating cell culture system to simulate the microgravity environment.The proliferation rate,acid resistance and biofilm formation of the strains were measured.The phenotypic changes of Staphylococcus aureus were evaluated.Transcriptome sequencing was used to detect the differentially expressed genes under simulated microgravity conditions and then they were compared with the phenotypes.Results Simulated microgravity resulted in a decrease in the proliferation rate of Staphylococcus aureus,an increase in the acid resistance,an enhancement in biofilm formation,and a down-regulation of alkalinity.There were a total of 172 differentially expressed genes in the simulated microgravity group(P<0.05,of which 20 were up-regulated and 152 were down-regulated),there were 36 differentially expressed genes related to nutrition metabolism,32 of which decreased in expression,and 1 was associated with decreased expression of alkali-resistant genes.Conclusion Simulated microgravity can cause the phenotype and corresponding genetic changes in Staphylococcus aureus,and the enhancement of biofilm formation may pose a potential threat to space flight.
引文
[1]Stewart LH,Trunkey D,Rebagliati GS.Emergency medicine in space[J].Journal of Emergency Medicine,2007,32(1):45.
[2]Rosenzweig JA,Ahmed S,John Eunson J,et al.Lowshear force associated with modeled microgravity and spaceflight does not similarly impact the virulence of notable bacterial pathogens[J].Applied Microbiology&Biotechnology,2014,98(21):8797-8807.
[3]Novikova ND.Review of the knowledge of microbial contamination of the Russian manned spacecraft[J].Microbial Ecology,2004,47(2):127-132.
[4]Novikova N,De BP,Poddubko S,et al.Survey of environmental biocontamination on board the International Space Station[J].Research in Microbiology,2006,157(1):5-12.
[5]Song B,Leff LG.Identification and characterization of bacterial isolates from the Mir space station[J].Microbiological Research,2005,160(2):111-117.
[6]Klaus DM,Howard HN.Antibiotic efficacy and microbial virulence during space flight[J].Trends in Biotechnology,2006,24(3):131-136.
[7]Horneck G,Klaus DM,Mancinelli RL.Space microbiology[J].Microbiology&Molecular Biology Reviews Mmbr,2010,74(1):121-156.
[8]Schwarz RP,Goodwin TJ,Wolf DA.Cell culture for three-dimensional modeling in rotating-wall vessels:An application of simulated microgravity[J].Journal of Tissue Culture Methods,1992,14(2):51-57.
[9]容丹,王佳平,王海立,等.模拟失重对大肠杆菌K12基因表达及表型的影响[J].微生物学通报,2017,44(5):1038-1046.Rong D,Wang JP,Wang HL,et al.Effect of simulated weightlessness on the expression and phenotype of Escherichia coli K12gene[J].Microbiology,2017,44(5):1038-1046.
[10]王海立.模拟微重力环境中肺炎克雷伯菌重要生理表型变化及其潜在机制研究[D].北京:中国人民解放军军事医学科学院,2016.Wang HL.Study on the physiological phenotypic changes of Klebsiella pneumoniae in simulated microgravity environment and its potential mechanism[D].Beijing:Chinese Academy of Military Medical Sciences,2016.
[11]尹焕才,薛小平,杨慧,等.模拟微重力环境下细菌生物学效应的初步研究[J].航天医学与医学工程,2009,22(5):341-346.Yin HC,Xue XP,Yang H,et al.Preliminary study on bacterial biological effects under simulated microgravity environment[J].Space Medicine&Medical Engineering,2009,22(5):341-346.
[12]Castro SL,Nelmangonzalez M,Nickerson CA,et al.Induction of attachment-independent biofilm formation and repression of Hfq expression by low-fluid-shear culture of Staphylococcus aureus[J].Applied&Environmental Microbiology,2011,77(18):6368-6378.
[13]Tixador R,Richoilley G,Gasset G,et al.Preliminary results of Cytos 2experiment[J].Acta Astronautica,1985,12(2):131-134.
[14]Hamilton-Miller JM,Shah S.Disorganization of cell division of methicillin-resistant Staphylococcus aureus by a component of tea(Camellia sinensis):A study by electron microscopy[J].Fems Microbiology Letters,1999,176(2):463.
[15]Sieradzki K,Pinho MG,Tomasz A.Inactivated pbp4in Highly glycopeptide-resistant laboratory mutants of Staphylococcus aureus[J].Journal of Biological Chemistry,1999,274(27):18942-18926.
[16]Sieradzki K,Tomasz A.Alterations of cell wall structure and metabolism accompany reduced susceptibility to vancomycin in an isogenic series of clinical isolates of Staphylococcus aureus[J].Journal of Bacteriology,2003,185(24):7103-7110.
[17]Kim W,Tengra FK,Young Z,et al.Spaceflight promotes biofilm formation by Pseudomonas aeruginosa[J].PloSOne,2013,8(4):e62437.
[18]Mauclaire L,Egli M.Effect of simulated microgravity on growth and production of exopolymeric substances of Micrococcus luteus space and earth isolates[J].Fems Immunology&Medical Microbiology,2010,59(3):350-356.
[19]Lynch SV,Mukundakrishnan K,Benoit MR,et al.Escherichia coli biofilms formed under low-shear modeled microgravity in a ground-based system[J].Applied&Environmental Microbiology,2006,72(12):7701-7710.
[20]Wilson JW,Nickerson CA.Microarray analysis identifies Salmonella genes belonging to the low-shear modeled microgravity regulon[J].Proceedings of the National Academy of Sciences,2002,99(21):13807-13812.
[21]Wilson JW,Ott CM,Quick L,et al.Media ion composition controls regulatory and virulence response of Salmonella in spaceflight[J].PloS One,2008,3(12):e3923.
[22]Rosado H,Doyle M,Hinds J,et al.Low-shear modelled microgravity alters expression of virulence determinants of Staphylococcus aureus[J].Acta Astronautica,2010,66(3-4):408-413.
[2]Rosenzweig JA,Ahmed S,John Eunson J,et al.Lowshear force associated with modeled microgravity and spaceflight does not similarly impact the virulence of notable bacterial pathogens[J].Applied Microbiology&Biotechnology,2014,98(21):8797-8807.
[3]Novikova ND.Review of the knowledge of microbial contamination of the Russian manned spacecraft[J].Microbial Ecology,2004,47(2):127-132.
[4]Novikova N,De BP,Poddubko S,et al.Survey of environmental biocontamination on board the International Space Station[J].Research in Microbiology,2006,157(1):5-12.
[5]Song B,Leff LG.Identification and characterization of bacterial isolates from the Mir space station[J].Microbiological Research,2005,160(2):111-117.
[6]Klaus DM,Howard HN.Antibiotic efficacy and microbial virulence during space flight[J].Trends in Biotechnology,2006,24(3):131-136.
[7]Horneck G,Klaus DM,Mancinelli RL.Space microbiology[J].Microbiology&Molecular Biology Reviews Mmbr,2010,74(1):121-156.
[8]Schwarz RP,Goodwin TJ,Wolf DA.Cell culture for three-dimensional modeling in rotating-wall vessels:An application of simulated microgravity[J].Journal of Tissue Culture Methods,1992,14(2):51-57.
[9]容丹,王佳平,王海立,等.模拟失重对大肠杆菌K12基因表达及表型的影响[J].微生物学通报,2017,44(5):1038-1046.Rong D,Wang JP,Wang HL,et al.Effect of simulated weightlessness on the expression and phenotype of Escherichia coli K12gene[J].Microbiology,2017,44(5):1038-1046.
[10]王海立.模拟微重力环境中肺炎克雷伯菌重要生理表型变化及其潜在机制研究[D].北京:中国人民解放军军事医学科学院,2016.Wang HL.Study on the physiological phenotypic changes of Klebsiella pneumoniae in simulated microgravity environment and its potential mechanism[D].Beijing:Chinese Academy of Military Medical Sciences,2016.
[11]尹焕才,薛小平,杨慧,等.模拟微重力环境下细菌生物学效应的初步研究[J].航天医学与医学工程,2009,22(5):341-346.Yin HC,Xue XP,Yang H,et al.Preliminary study on bacterial biological effects under simulated microgravity environment[J].Space Medicine&Medical Engineering,2009,22(5):341-346.
[12]Castro SL,Nelmangonzalez M,Nickerson CA,et al.Induction of attachment-independent biofilm formation and repression of Hfq expression by low-fluid-shear culture of Staphylococcus aureus[J].Applied&Environmental Microbiology,2011,77(18):6368-6378.
[13]Tixador R,Richoilley G,Gasset G,et al.Preliminary results of Cytos 2experiment[J].Acta Astronautica,1985,12(2):131-134.
[14]Hamilton-Miller JM,Shah S.Disorganization of cell division of methicillin-resistant Staphylococcus aureus by a component of tea(Camellia sinensis):A study by electron microscopy[J].Fems Microbiology Letters,1999,176(2):463.
[15]Sieradzki K,Pinho MG,Tomasz A.Inactivated pbp4in Highly glycopeptide-resistant laboratory mutants of Staphylococcus aureus[J].Journal of Biological Chemistry,1999,274(27):18942-18926.
[16]Sieradzki K,Tomasz A.Alterations of cell wall structure and metabolism accompany reduced susceptibility to vancomycin in an isogenic series of clinical isolates of Staphylococcus aureus[J].Journal of Bacteriology,2003,185(24):7103-7110.
[17]Kim W,Tengra FK,Young Z,et al.Spaceflight promotes biofilm formation by Pseudomonas aeruginosa[J].PloSOne,2013,8(4):e62437.
[18]Mauclaire L,Egli M.Effect of simulated microgravity on growth and production of exopolymeric substances of Micrococcus luteus space and earth isolates[J].Fems Immunology&Medical Microbiology,2010,59(3):350-356.
[19]Lynch SV,Mukundakrishnan K,Benoit MR,et al.Escherichia coli biofilms formed under low-shear modeled microgravity in a ground-based system[J].Applied&Environmental Microbiology,2006,72(12):7701-7710.
[20]Wilson JW,Nickerson CA.Microarray analysis identifies Salmonella genes belonging to the low-shear modeled microgravity regulon[J].Proceedings of the National Academy of Sciences,2002,99(21):13807-13812.
[21]Wilson JW,Ott CM,Quick L,et al.Media ion composition controls regulatory and virulence response of Salmonella in spaceflight[J].PloS One,2008,3(12):e3923.
[22]Rosado H,Doyle M,Hinds J,et al.Low-shear modelled microgravity alters expression of virulence determinants of Staphylococcus aureus[J].Acta Astronautica,2010,66(3-4):408-413.