不同寡营养条件下饥饿期粪肠球菌基因转录水平的体外研究
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摘要
目的:在体外模拟根管致病压力条件下,探究不同寡营养条件对饥饿期粪肠球菌基因转录水平的影响。方法:将粪肠球菌标准菌株(ATCC 29212)诱导至饥饿期后培养于不同寡营养条件下(0%、0.05%、0.15%和0.25%葡萄糖浓度的培养液),1周后对其基因转录水平及代谢通路进行分析。结果:转录组结果显示,与0.25%相比,0.15%葡萄糖浓度基因转录水平没有明显变化,0.05%及0%葡萄糖浓度基因转录水平变化明显,以0%葡萄糖浓度变化最为显著,在GO功能上,显著富集于代谢进程及催化活性,在KEGG功能上并无显著差异,排名前3的代谢通路有ABC转运器、磷酸转移酶系统、氨基酸生物合成。结论:寡营养条件下,粪肠球菌基因转录水平的变化可能和其在恶劣生存条件下仍能生存和致病有关,在本研究基础上将进一步探究粪肠球菌相关功能基因和调控通路在致病过程中的机制。
Objective:This study aims to investigate the in vitro effects of different oligotrophic conditions on the transcriptional level of Enterococcus faecalis(E.faecalis)during starvation phase under simulated root canal pressure. Methods:The E.faecalis(ATCC29212)was induced to starvation phase and cultured under different oligotrophic conditions(TSB with 0%,0.05%,0.15%,and 0.25%glucose),gene transcription levels and metabolic pathways were measured after one week. Results:The results showed that there was no significant change in the gene transcription level in the 0.15% glucose group compared with the 0.25% group. The gene transcription level in the 0.05% glucose group and particular in the 0% glucose group changed significantly. In the 0% glucose group,GO function mostly involved in metabolic processes and catalytic activity,and there was no significant difference in KEGG pathway.The top three metabolic pathways were ABC transporter,PTS system and amino acid biosynthesis. Conclusion:The changes in gene transcription level of E.faecalis under oligotrophic conditions may be related to the survival and pathogenicity of E.faecalis under starvation conditions. We will further explore the mechanism of related functional genes and regulatory pathways of E.faecalis in the pathogenesis of the disease on the basis of this study.
Objective:This study aims to investigate the in vitro effects of different oligotrophic conditions on the transcriptional level of Enterococcus faecalis(E.faecalis)during starvation phase under simulated root canal pressure. Methods:The E.faecalis(ATCC29212)was induced to starvation phase and cultured under different oligotrophic conditions(TSB with 0%,0.05%,0.15%,and 0.25%glucose),gene transcription levels and metabolic pathways were measured after one week. Results:The results showed that there was no significant change in the gene transcription level in the 0.15% glucose group compared with the 0.25% group. The gene transcription level in the 0.05% glucose group and particular in the 0% glucose group changed significantly. In the 0% glucose group,GO function mostly involved in metabolic processes and catalytic activity,and there was no significant difference in KEGG pathway.The top three metabolic pathways were ABC transporter,PTS system and amino acid biosynthesis. Conclusion:The changes in gene transcription level of E.faecalis under oligotrophic conditions may be related to the survival and pathogenicity of E.faecalis under starvation conditions. We will further explore the mechanism of related functional genes and regulatory pathways of E.faecalis in the pathogenesis of the disease on the basis of this study.
引文
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[2]Jr SJ,R??as IN. Polymerase chain reaction-based analysis of microorganisms associated with failed endodontic treatment[J]. Oral Surg Oral Med Oral Pathol Oral Radiol Endod,2004,97(1):85
[3]Zhang C,Du J,Peng Z. Correlation between enterococcus faecalis and persistent intraradicular infection compared with primary intraradicular infection:A systematic review[J]. J Endod,2015,41(8):1207-1213
[4]Leron K,Mor S,Shaul B,et al. Phage therapy against Enterococcus faecalisin dental root canals[J]. J Oral Microbiol,2016,8(1):32157
[5]Heim S,Lleo MDM,Bonato B,et al. The viable but nonculturable state and starvation are different stress responses of enterococcus faecalis,as determined by proteome analysis[J]. J Bacteriol,2002,184(23):6739-6745
[6]冉淑君,鄂佳,朱彩莲,等.不同理化条件对粪肠球菌生长周期及生物膜形成能力的影响[J].中华口腔医学杂志,2013,48(9):529-534
[7]Ran S,Jiang W,Zhu C,et al. Exploration of the mechanisms of biofilm formation by Enterococcus faecalis in glucose starvation environments[J]. Aust Dent J,2015,60(2):143-153
[8]周如玉,平逸帆,张元,等.口腔综合治疗台水路系统中假单胞菌属污染状况的检测[J].南京医科大学学报(自然科学版),2017,37(4):452-457
[9]Aguirre-Santoro J,Michelangeli FA,Stevenson DW. Molecular phylogenetics of the Ronnbergia Alliance(Bromeliaceae,Bromelioideae)and insights into their morphological evolution[J]. Mol Phylogenet Evol,2016,100(1):1-20
[10]O’brien S,Hodgson DJ,Buckling A. The interplay between microevolution and community structure in microbial populations[J]. Curr Opin Biotechnol,2013,24(4):821-825
[11]Langmead B,Trapnell C,Pop M,et al. Ultrafast and memory-efficient alignment of short DNA sequences to the human genome[J]. Genome Biol,2009,10(3):R25
[12]Camacho C,Coulouris G,Avagyan V. BLAST plus:Architecture and applications[J]. BMC Bioinformatics,2009,10(1):421
[13]Trapnell C,Williams BA,Pertea G,et al. Transcript assembly and quantification by RNA-Seq reveals unannotated transcripts and isoform switching during cell differentiation[J]. Nat Biotechnol,2010,28(5):511-515
[14]Gao Y,Jiang X,Lin D,et al. The Starvation resistance and biofilm formation of enterococcus faecalis in coexistence with Candida albicans,Streptococcus gordonii,Actinomyces viscosus,or Lactobacillus acidophilus[J]. J Endod,2016,42(8):1233-1238
[15]Pillai SK,Sakoulas G,Eliopoulos GM,et al. Effects of glucose on fsr-mediated biofilm formation in Enterococcus faecalis[J]. J Infect Dis,2004,190(5):967-970
[16]刘红艳,韦曦,凌均棨.饥饿状态粪肠球菌生物膜胞外多糖的合成能力[J].中华口腔医学研究杂志(电子版),2012,6(5):16-20
[17]Cascorbi I,Haenisch S. Pharmacogenetics of ATP-binding cassette transporters and clinical implications[J]. Methods Mol Biol,2010,596:95-121
[18]Jr MHS,Reizer J. The bacterial phosphotransferase system:new frontiers 30 years later[J]. Mol Microbiol,2010,13(5):755-764
[19]Morschh?¤User J. The development of fluconazole resistance in Candida albicans-an example of microevolution ofafungalpathogen[J].JMicrobiol,2016,54(3):192-201
[20]Stavros B,Gili ZS,Eran E. Use of metatranscriptomics in microbiome research[J]. Bioinform Biol Insights,2016,10(10):19-25
[21]Choi SC. On the study of microbial transcriptomes using second-and third-generation sequencing technologies[J].J Microbiol,2016,54(8):527-536
[22]Aseev LV,Koledinskaya LS,Boni IV. Regulation of ribosomal protein operons rplM-rpsI,rpmB-rpmG,rplU-rpmA at the transcriptional and translational levels[J]. J Bacteriol,2016,198(18):2494-2502
[23]Ehrnthaler M,Scharff LB,Fleischmann TT,et al. Synthetic lethality in the tobacco plastid ribosome and its rescue at elevated growth temperatures[J]. Plant Cell,2014,26(2):765-776
[24]Hennig S,Ziebuhr W. Characterization of the Transposase encoded by IS256,the prototype of a major family of bacterial insertion sequence elements[J]. J Bacteriol,2010,192(16):4153-4163
[25]Perez M,Callesenríquez M,Del RB,et al. IS256 abolishes gelatinase activity and biofilm formation in a mutant of the nosocomial pathogen Enterococcus faecalis V583[J].Can J Microbiol,2015,61(7):517-519