产膜表皮葡萄球菌LuxS/AI-2型密度感应系统AI-2的活性及苦参碱对其的影响
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摘要
【目的】研究产膜表皮葡萄球菌生物被膜菌和液相浮游菌LuxS/AI-2型密度感应系统中AI-2信号分子的活性及苦参碱对其的影响。【方法】绘制产膜表皮葡萄球菌ATCC35984液相浮游菌及生物被膜菌的生长曲线;利用哈维氏弧菌BB170(Vibrio harveyi BB170)作为报告菌株检测表皮葡萄球菌AI-2信号分子的活性;用荧光定量PCR法检测luxS基因的转录水平;用苦参碱对产膜表皮葡萄球菌ATCC35984进行处理,检测其AI-2信号分子活性的变化情况。【结果】在产膜表皮葡萄球菌液相浮游菌生长过程中,AI-2信号分子与luxS基因的相对表达量都是在对数生长期达到最大值,而后逐渐降低,与液相浮游菌生长曲线基本一致。在产膜表皮葡萄球菌生物被膜菌生长过程中,AI-2信号分子活性曲线与其生物被膜生长曲线变化趋势一致,但luxS基因的相对表达量曲线与生物被膜生长曲线变化趋势相反。中药苦参碱对产膜表皮葡萄球菌液相浮游菌和生物被膜菌AI-2信号分子活性有抑制作用,并且对液相浮游菌AI-2信号分子活性的抑制作用强于生物被膜菌。【结论】AI-2信号分子活性和luxS基因的相对表达量与产膜表皮葡萄球菌生物被膜菌及其液相浮游菌的生长相关;苦参碱能够显著下调产膜表皮葡萄球菌生物被膜菌和液相浮游菌的AI-2信号分子活性。
【Objective】This study investigated the activity of AI-2signaling molecules in Staphylococ cusepidermidisplanktonic bacteria and biofilm bacteria and the effect of matrine.【Method】The growth curve of Staphylococcus epidermidis planktonic bacteria and biofilm bacteria was drawn and AI-2concentration was measured using the V.harveyi bioluminescence assay.The transcription level of luxSgene was analyzed using real-time PCR.The AI-2signaling molecules of Staphylococcusepidermidis ATCC35984 were treated with matrine and the changes of AI-2signaling molecules activity were determined.【Result】In Staphylococcus epidermidis planktonic bacteria growth process,AI-2signaling molecule activity and the relative expression quantity of luxSgene reached the maximum in the logarithmic phase before decreasing gradually.Their change curves were similar with planktonic growth curve.However,in the growth processof Staphylococcus epidermidis biofilm bacteria,the activity trend was similar with the biofilm growth.The relative expression quantity of luxSgene had an opposite trend to biofilm growth.Matrine had inhibitory effect on AI-2signaling molecule activity in planktonic and biofilm bacteria,and the inhibitory effect was stronger to planktonic bacteria.【Conclusion】AI-2signaling molecule activity and relative expression quantity of luxSgene highly correlated with the growth of Staphylococcus epidermidis and its biofilm formation.Matrine significantly reduced the AI-2signaling molecule activity of Staphylococcus epidermidis.
【Objective】This study investigated the activity of AI-2signaling molecules in Staphylococ cusepidermidisplanktonic bacteria and biofilm bacteria and the effect of matrine.【Method】The growth curve of Staphylococcus epidermidis planktonic bacteria and biofilm bacteria was drawn and AI-2concentration was measured using the V.harveyi bioluminescence assay.The transcription level of luxSgene was analyzed using real-time PCR.The AI-2signaling molecules of Staphylococcusepidermidis ATCC35984 were treated with matrine and the changes of AI-2signaling molecules activity were determined.【Result】In Staphylococcus epidermidis planktonic bacteria growth process,AI-2signaling molecule activity and the relative expression quantity of luxSgene reached the maximum in the logarithmic phase before decreasing gradually.Their change curves were similar with planktonic growth curve.However,in the growth processof Staphylococcus epidermidis biofilm bacteria,the activity trend was similar with the biofilm growth.The relative expression quantity of luxSgene had an opposite trend to biofilm growth.Matrine had inhibitory effect on AI-2signaling molecule activity in planktonic and biofilm bacteria,and the inhibitory effect was stronger to planktonic bacteria.【Conclusion】AI-2signaling molecule activity and relative expression quantity of luxSgene highly correlated with the growth of Staphylococcus epidermidis and its biofilm formation.Matrine significantly reduced the AI-2signaling molecule activity of Staphylococcus epidermidis.
引文
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[11]Sun Z,He X,Brancaccion V F,et al.Bifidobacteria exhibit LuxS-dependent autoinducer 2activity and biofilm formation[J].PLoS One,2014,9(2):e88260.
[12]白灏,韩先干,刘蕾,等.影响禽致病性大肠杆菌信号分子AI-2产生的因素分析[J].中国农业科学,2013,46(15):3220-3226.Bai H,Han X G,Liu L,et al.Analysis of factors affecting the activity of AI-2of avian pathogenic Escherichia coli[J].Science Agricultura Sinica,2013,46(15):3220-3226.
[13]周蕾.鼠疫耶尔森氏菌密度感应系统研究[D].北京:军事医学科学院微生物流行病研究所,2007.Zhou L.The research of quorum sensing system in Yersinia pestis[D].Beijing:Institute of Microbiology and Epidemiology,Academy of Military Medical Science,2007.
[14]Jorge E V,Herbert P L,Kunkel R M,et al.The LuxS-dependent quorum-sensing system regulates early biofilm formation by Streptococcus pneumoniae strain D39[J].Infection and Immunity,2011,79(10):4050-4060.
[15]喻华英,李启峰.大肠杆菌生物膜的筛选及生长曲线测定[J].中国农学通报,2014,30(5):23-27.Yu H Y,Li Q F.Screening biofilm and determination growth biofilm of curve in Escherichia coil[J].Chinese Agricultural Science Bulletin,2014,30(5):23-27.
[16]Xavier K B,Bassler B L.LuxS quorum sensing:more than just a numbers game[J].Current Opinion in Microbiology,2003,6(2):191-197.
[17]Xu L,Li H,Vuong C,et al.Role of the luxS quorum-sensing system in biofilm formation and virulence of Staphylococcus epidermidis[J].Infection and Immunity,2006,74(1):488-496.
[18]Husain F M,Ahmad I,Khan M S,et al.Sub-MICs of Mentha piperita essential oil and menthol inhibits AHL mediated quorum sensing and biofilm of Gram-negative bacteria[J].Frontiers in Microbiology,2015,6:420.
[2]刘琳,谭小娟.细菌群体感应与细菌生物被膜形成之间的关系[J].微生物学报,2012,52(3):271-278.Liu L,Tan X J.Relationship between bacterial quorum sensing and biofilm formation-A review[J].Acta Microbiological Sinica,2012,52(3):271-278.
[3]Chu W H,Zhou S H,Jiang Y,et al.Effect of traditional chinese herbal medicine with antiquorum sensing activity on Pseudomonas aeruginosa[J].Evidence-Based Complementary and Alternative Medicine,2013(2):1-7.
[4]Manmohit K,Vivek K Y,Pradeep K S,et al.Effect of cinnamon oil on quorum sensing-controlled virulence factors and biofilm formation in Pseudomonas aeruginosa[J].PLoS One,2015,10(8):e0135495.
[5]Deryabin D G,Tolmacheva A A.Antibacterial and anti-quorum sensing molecular composition derived from quercus cortex(Oak bark)extract[J].Molecules,2015,20(9):17093-17108.
[6]官妍,马越,程惠娟,等.苦参碱对表皮葡萄球菌生物被膜作用初探[J].微生物学通报,2011,38(1):91-96.Guan Y,Ma Y,Cheng H J,et al.Preliminary study of effects of matrine on Staphylococcus epidermidis biofilms[J].Microbiology China,2011,38(1):91-96.
[7]Yu D,Zhao L,Xue T,et al.Staphylococcus aureus autoinducer-2quorum sensing decreases biofilm formation in an icaR dependent manner[J].BioMed Central Microbiol,2012,12:288-295.
[8]徐玉善.luxS在葡萄糖诱导的生物材料表面细菌生物膜形成与分离中的作用[D].昆明:昆明医科大学,2012.Xu Y S.Role of luxS in the formation and detachment of biofilm which be induced by glucose on the surface of biomaterials of bacteria[D].Kunming:Kunming Medical University,2012.
[9]朱娇玲,乔霞,李雪,等.奶牛乳腺炎表皮葡萄球菌耐药性及生物被膜形成相关基因的研究[J].内蒙古农业大学学报,2015,36(2):1-8.Zhu J L,Qiao X,Li X,et al.Drug resistance and biofilm formation of associated genes involved in Staphylococcus epidermidis isolates from bovine mastitis[J].Journal of Inner Mongolia Agricultural University,2015,36(2):1-8.
[10]Siller M,Janapatla R P,Pirzada Z A,et al.Functional analysis of the group A streptococcal luxS/AI-2system in metabolism,adaptation to stress and interaction with host cells[J].BioMed Central Microbiol,2008,8:188-205.
[11]Sun Z,He X,Brancaccion V F,et al.Bifidobacteria exhibit LuxS-dependent autoinducer 2activity and biofilm formation[J].PLoS One,2014,9(2):e88260.
[12]白灏,韩先干,刘蕾,等.影响禽致病性大肠杆菌信号分子AI-2产生的因素分析[J].中国农业科学,2013,46(15):3220-3226.Bai H,Han X G,Liu L,et al.Analysis of factors affecting the activity of AI-2of avian pathogenic Escherichia coli[J].Science Agricultura Sinica,2013,46(15):3220-3226.
[13]周蕾.鼠疫耶尔森氏菌密度感应系统研究[D].北京:军事医学科学院微生物流行病研究所,2007.Zhou L.The research of quorum sensing system in Yersinia pestis[D].Beijing:Institute of Microbiology and Epidemiology,Academy of Military Medical Science,2007.
[14]Jorge E V,Herbert P L,Kunkel R M,et al.The LuxS-dependent quorum-sensing system regulates early biofilm formation by Streptococcus pneumoniae strain D39[J].Infection and Immunity,2011,79(10):4050-4060.
[15]喻华英,李启峰.大肠杆菌生物膜的筛选及生长曲线测定[J].中国农学通报,2014,30(5):23-27.Yu H Y,Li Q F.Screening biofilm and determination growth biofilm of curve in Escherichia coil[J].Chinese Agricultural Science Bulletin,2014,30(5):23-27.
[16]Xavier K B,Bassler B L.LuxS quorum sensing:more than just a numbers game[J].Current Opinion in Microbiology,2003,6(2):191-197.
[17]Xu L,Li H,Vuong C,et al.Role of the luxS quorum-sensing system in biofilm formation and virulence of Staphylococcus epidermidis[J].Infection and Immunity,2006,74(1):488-496.
[18]Husain F M,Ahmad I,Khan M S,et al.Sub-MICs of Mentha piperita essential oil and menthol inhibits AHL mediated quorum sensing and biofilm of Gram-negative bacteria[J].Frontiers in Microbiology,2015,6:420.