抗菌肽抑制细菌生物膜的研究现状

随着抗生素耐药菌株的大量出现,抗菌肽的使用为临床感染的治疗提供了一种新的思路和手段。抗菌肽作为一种新型的抗菌制剂应用于抑制和杀灭细菌生物膜的过程中发挥了独特的优势,也存在明显的缺陷。本文将对抗菌肽作用于细菌生物膜的研究现状,在应用中目前存在的问题及可能采取的解决办法进行综述。     

大肠杆菌鞭毛调控基因flhDC与生物材料植入感染的研究进展

随着生物材料被广泛应用于临床,生物材料植入感染成为令人棘手的常见医院内感染,有报道占院内感染的50%,大肠杆菌是临床以生物材料为中心感染的优势菌种。生物材料表面的细菌生物膜使其膜内细菌能有效抵御抗生素治疗和机体的防御反应,是导致生物材料为中心感染难以控制的根源。大肠杆菌的运动性与细菌生物膜形成密切相关,鞭毛是大肠杆菌的运动器官,鞭毛的生成需要三级基因的表达,操纵子flh DC编码鞭毛生成的一级主调控基因。我们推测:"鞭毛调控基因flh DC的表达→鞭毛的生成→细菌的运动性→细菌生物膜形成"之间存在着一一对应的关系,这为临床防治生物材料植入感染提供新思路。本文就以大肠杆菌鞭毛调控基因flh DC与生物材料植入感染做一简要综述。     

细菌生物膜与耐药性相关性研究进展

细菌生物膜是一种包裹于细胞外多聚物基质中不可逆的黏附于非生物或生物表面的微生物细胞菌落.生物膜状态下的细菌相对其浮游状态具有显著增强的耐药性,对人及动物细菌性感染具有重要研究价值.然而尽管动物细菌耐药性被广泛报道,却很少涉及细菌生物膜与其之间的相关性,本文综述了细菌生物膜的耐药机制并探讨了细菌生物膜与动物源性细菌耐药性的关系,可作为研究细菌耐药性及控制动物产品安全的参考.     

细菌生物膜形成与慢性难愈性创面相关性的研究进展

感染是影响慢性难愈性创面愈合最常见的原因,由于多种细菌混合感染、耐药性产生、生物膜的形成使其治疗成为难题.其中,细菌生物膜形成是导致创面的难以愈合的重要因素之一.本文就慢性难愈合创面中细菌生物膜的形成机制、特征、生态学、对伤口愈合的影响以及可能的治疗对策等作一综述.     

细菌群体感应对细菌生物膜形成与调控的研究进展

细菌群体感应(quorum sensing,QS)是细菌细胞之间通过感受自诱导物来调控细菌群体行为的现象。细菌生物膜(bacterial biofilm,BBF)是细菌在生长过程中为适应生存环境而吸附于惰性或活性材料表面形成的一种与浮游细胞相应的生物被膜,其结构包括细菌和自身分泌的细胞外基质(extracelluar polymeric substance,EPS)。生物膜可以保护细菌免于外界恶劣环境的影响,增加其抗药性,为细菌生长提供天然的有利屏障。细菌群体感应是细菌调控生物膜形成的重要机制之一,本文中,笔者综述了细菌群体感应在生物膜生成过程中的调控作用,分析了细菌生物膜抑制与降解策略,旨在为医药与食品工业中降低细菌耐药性和细菌抗性提供理论依据。     

铰链结构对抗菌肽生物活性的影响及在分子设计中的应用研究进展

铰链结构,又称铰链区或转角,是部分抗菌肽序列中存在的一种特殊结构。但目前抗菌肽结构的研究多集中于标准的α-螺旋和β-折叠二级结构,对于铰链结构及其作用总结较少。铰链结构对抗菌肽生物活性有重要影响,主要原因是铰链结构能够提高抗菌肽的结构灵活性,促进其对细菌细胞膜的破坏作用或与胞内作用靶点的结合效率,进而提高抗菌肽的抗菌活性。同时,降低的抗菌肽结构刚性,消减了抗菌肽对真核细胞的毒性。文中结合了笔者课题组相关工作,就铰链结构特点、对抗菌肽生物活性的影响以及在抗菌肽分子设计方面的应用进行了综述,以期为新型抗菌肽的设计和开发提供参考。     

连翘苷和黄芩苷对表皮葡萄球菌生物膜抑制作用的研究

目的通过中药有效成分连翘苷和黄芩苷分别对表皮葡萄球菌生物膜抑制作用的研究,为表皮葡萄球菌生物膜引起的相关感染提供新的治疗途径。方法体外构建表皮葡萄球菌生物膜,XTT减低法评价连翘苷、黄芩苷对表皮葡萄球菌初始黏附及生物膜内细菌代谢的影响,显微镜下观察用药后表皮葡萄球菌生物膜形态和结构改变。结果连翘苷和黄芩苷对表皮葡萄球菌生物膜的早期黏附均无抑制作用;连翘苷对表皮葡萄球菌生物膜菌的SMIC50为31.25μg/ml,而黄芩苷对表皮葡萄球菌生物膜菌的代谢无影响;在显微镜下观察,连翘苷使部分表皮葡萄球菌被膜的形态发生改变,而黄芩苷对其形态影响不显著。结论连翘苷对表皮葡萄球菌生物膜的初始黏附阶段无抑制作用,对生物膜菌的代谢和生物膜形态均有显著影响;黄芩苷对表皮葡萄球菌生物膜无显著作用。     

细菌对抗菌肽的耐受机制

抗菌肽广谱、高特异、高生物活性等特点决定其具极大的临床应用潜力,然而抗菌肽的耐受是其药物开发必须重视和亟待克服的问题。从生物学的观点看,部分细菌可以产生抗菌肽,其必定存在逃避自身抗菌肽作用的机制;从进化的观点看,宿主和病原体之间是相互抑制、相互逃避、相互适应的关系,细菌在漫长的进化中会形成应对抗菌肽的特殊机制。抗菌肽对细菌存在多种作用机制,其核心是依赖于与细胞膜相互作用或进入细胞,进而改变膜完整性或干扰胞内生理生化反应导致细菌死亡;而细菌通过减弱抗菌肽结合、降低抗菌肽有效浓度等方式产生对抗菌肽的耐受。这些耐受机制也为抗菌肽类药物开发提供重要的启示。     

抗菌肽对细菌杀伤作用的分子机制

抗菌肽是一类新型的抗菌物质,从最低等的生物病毒、细菌到高等的动植物都有广泛分布. 以往的研究主要集中于抗菌肽对细菌细胞膜的作用机制,已经构建了三种作用模式. 但近几年的研究表明,很多抗菌肽都能有效地穿过细菌的细胞膜,直接与胞内分子相互作用,并不引起膜的破裂. 抗菌肽根据其结构特点有着多种杀菌穿膜的机制,其后分别与胞内的靶分子如核酸,蛋白质,信号转导通路等互相作用,最终实现对细菌的杀伤作用.     

噬菌体酶类降解细菌生物膜的研究进展

细菌生物膜(bacterial biofilm, BF)是细菌产生耐药性的重要原因之一,给抗生素治疗细菌感染带来极大困难。生物膜主要由细菌菌体和胞外聚合物质(extracellular polymeric substance, EPS)构成,其中EPS成分复杂,主要包括多糖(polysaccharides)、蛋白质、核酸和脂类等。噬菌体是一类特异性侵染细菌且以细菌为宿主的病毒,可编码多种酶类,在感染过程中打破细菌的防御屏障。其中,解聚酶(depolymerase)可以降解EPS中的多糖成分;溶素(lysin)可以降解肽聚糖,破坏生物膜的完整结构,降低细菌的耐药性。结合噬菌体本身的特异性裂解作用,可以协同改善耐药菌感染的抗生素治疗效果。现就噬菌体编码的解聚酶与溶素对细菌生物膜的影响作一阐述。     

c-di-AMP调控细菌生物被膜的形成

细菌生物被膜是细菌持续性致病的重要机制。研究细菌生物被膜的形成和发展可为顽固性细菌感染防治提供新的思路与策略。环二腺苷酸c-di-AMP(Cyclic diadenosine monophosphate)是继c-di-GMP之后在细菌中新发现的一种核苷酸第二信使分子。研究发现,c-di-AMP参与调节细菌多种生理功能,包括细菌生长代谢、生物被膜形成、细胞壁的合成以及细菌毒力因子等。本文综述了c-di-AMP参与调控细菌生物被膜形成的不同方式及其分子机制。鉴于c-di-AMP在调控细菌生物被膜中的重要性,其可作为抗细菌生物被膜感染新药研发的潜在靶点。     

Fusobacterium nucleatum in periodontal health and disease

The pathogenesis of periodontitis involves the interplay of microbiota present in the subgingival plaque and the host responses. Inflammation and destruction of periodontal tissues are considered to result from the response of a susceptible host to a microbial biofilm containing gram-negative pathogens. Antimicrobial peptides are important contributors to maintaining the balance between health and disease in this complex environment. These include several salivary antimicrobial peptides such as β-defensins expressed in the epithelium and LL-37 expressed in both epithelium and neutrophils. Among gram-negative bacteria implicated in periodontal diseases, Fusobacterium nucleatum, is one of the most interesting. This review will focus on expression, function, regulation and functional efficacy of antimicrobial peptides against F. nucleatum. We are looking for how the presence of F. nucleatum induces secretion of peptides which have an impact on host cells and modulate immune response.     

细菌生物被膜基质的研究进展

细菌生物被膜(biofilm)附着在生物或者非生物表面,由细菌及其分泌的糖、蛋白质和核酸等多种基质组成的细菌群落,是造成病原细菌持续性感染、毒力和耐药性的重要原因之一.细菌的生物被膜基质由复杂的胞外聚合物(extracellular polymeric substances,EPS)构成,影响生物被膜的结构和功能.本文...     

Bacterial envelope stress responses: Essential adaptors and attractive targets

Millions of deaths a year across the globe are linked to antimicrobial resistant infections. The need to develop new treatments and repurpose of existing antibiotics grows more pressing as the growing antimicrobial resistance pandemic advances. In this review article, we propose that envelope stress responses, the signaling pathways bacteria use to recognize and adapt to damage to the most vulnerable outer compartments of the microbial cell, are attractive targets. Envelope stress responses (ESRs) support colonization and infection by responding to a plethora of toxic envelope stresses encountered throughout the body; they have been co-opted into virulence networks where they work like global positioning systems to coordinate adhesion, invasion, microbial warfare, and biofilm formation. We highlight progress in the development of therapeutic strategies that target ESR signaling proteins and adaptive networks and posit that further characterization of the molecular mechanisms governing these essential niche adaptation machineries will be important for sparking new therapeutic approaches aimed at short-circuiting bacterial adaptation.     

Biofilm penetration and disinfection efficacy of alkaline hypochlorite and chlorosulfamates

AIMS: The purpose of this study was to compare the efficacy, in terms of bacterial biofilm penetration and killing, of alkaline hypochlorite (pH 11) and chlorosulfamate (pH 5.5) formulations. METHODS AND RESULTS: Two species biofilms of Pseudomonas aeruginosa and Klebsiella pneumoniae were grown by flowing a dilute medium over inclined stainless steel slides for 6 d. Microelectrode technology was used to measure concentration profiles of active chlorine species within the biofilms in response to treatment at a concentration of 1000 mg total chlorine l(-1). Chlorosulfamate formulations penetrated biofilms faster than did hypochlorite. The mean penetration time into approximately 1 mm-thick biofilms for chlorosulfamate (6 min) was only one-eighth as long as for the same concentration of hypochlorite (48 min). Chloride ion penetrated biofilms rapidly (5 min) with an effective diffusion coefficient in the biofilm that was close to the value for chloride in water. Biofilm bacteria were highly resistant to killing by both antimicrobial agents. Biofilms challenged with 1000 mg l(-1) alkaline hypochlorite or chlorosulfamate for 1 h experienced 0.85 and 1.3 log reductions in viable cell numbers, respectively. Similar treatment reduced viable numbers of planktonic bacteria to non-detectable levels (log reduction greater than 6) within 60 s. Aged planktonic and resuspended laboratory biofilm bacteria were just as susceptible to hypochlorite as fresh planktonic cells. CONCLUSION: Chlorosulfamate transport into biofilm was not retarded whereas hypochlorite transport clearly was retarded. Superior penetration by chlorosulfamate was hypothesized to be due to its lower capacity for reaction with constituents of the biofilm. Poor biofilm killing despite direct measurement of effective physical penetration of the antimicrobial agent into the biofilm demonstrates that bacteria in the biofilm are protected by some mechanism other than simple physical shielding by the biofilm matrix. SIGNIFICANCE AND IMPACT OF THE STUDY: This study lends support to the theory that the penetration of antimicrobial agents into microbial biofilms is controlled by the reactivity of the antimicrobial agent with biofilm components. The finding that chlorine-based biocides can penetrate, but fail to kill, bacteria in biofilms should motivate the search for other mechanisms of protection from killing by antimicrobial agents in biofilms.     

Antimicrobial peptide control of pathogenic microorganisms of the oral cavity: a review of the literature

Antimicrobial peptides, molecules produced in many different organisms, have high biocidal activity against several microorganisms. However, several questions about these molecules remain unclear. Therefore, this report details a systematic survey of the literature on the use of antimicrobial peptides against oral pathogens and indicates which peptides and microorganisms are most extensively studied. Articles were located using the PubMed and Science Direct databases with the following inclusion criteria: publication date between 2002 and 2011; keywords "biofilm OR biological film OR biological layer OR bacterial growth" AND "peptide" AND "oral cavity OR mouth OR buccal mucosa OR oral mucosa OR mouth mucosa"; and abstract in English. A total of 73 articles were selected after refinement of the data. An increase in publications focusing on the use of antimicrobial peptides against oral microorganisms was observed. In addition, the peptides produced by cells of the oral mucosa (defensins, LL-37 and histatins) as well as Streptococcus mutans (among cariogenic bacteria) and Porphyromonas gingivalis (among periodontal bacteria) were the most studied subjects. It was concluded that the use of antimicrobial peptides as a tool for microbial control is of increasing importance, likely due to its widespread use, mechanism of action, and low rates of bacterial resistance.     

细菌抗菌肽耐药机制研究进展

细菌对传统抗生素的耐药程度十分严重,寻找克服耐药性的新型抗菌药物已成为当务之急。抗菌肽(antimicrobial peptides,AMPs)是当下较有前景的抗菌药物之一。虽然通常认为,AMPs优先攻击细胞膜的特点使其不会引起广泛的耐药性,但其对特定靶标的识别能力仍为基因突变和细菌耐药性的产生提供了可能。此外,一些细菌还显示出了抵御宿主AMPs的杀伤作用并与宿主细胞共存的能力,相应的细菌防御机制也使其对治疗性AMPs产生抗性,这种交叉抗性近年来也备受关注。这些耐药现象的发现均对AMPs的开发提出了新挑战。本综述就细菌对AMPs耐药的分子机制进行了研究进展的总结,并且对治疗性AMPs与宿主防御肽交叉抗性的相关机制研究进行了归纳,以期寻求新的对抗耐药性的策略。     

Effects of side group functionality and molecular weight on the activity of synthetic antimicrobial polypeptides

The rapid emergence of antibiotic-resistant bacteria along with increasing difficulty in biofilm treatment has caused an immediate need for the development of new classes of antimicrobial therapeutics. We have developed a library of antimicrobial polypeptides, prepared by the ring-opening polymerization of γ-propargyl-L-glutamate N-carboxyanhydride and the alkyne-azide cycloaddition click reaction, which mimic the favorable characteristics of naturally occurring antimicrobial peptides (AmPs). AmPs are known not to cause drug resistance as well as prevent bacteria attachment on surfaces. The ease and scale of synthesis of the antimicrobial polypeptides developed here are significantly improved over the traditional Merrifield synthetic peptide approaches needed for naturally occurring antimicrobial peptides and avoids the unique challenges of biosynthetic pathways. The polypeptides range in length from 30 to 140 repeat units and can have varied side group functionality, including primary, secondary, tertiary, and quaternary amines with hydrocarbon side chains ranging from 1 to 12 carbons long. Overall, we find these polypeptides to exhibit broad-spectrum activity against both Gram positive and Gram negative bacteria, namely, S. aureus and E. coli , while having very low hemolytic activity. Many of the polypeptides can also be used as surface coatings to prevent bacterial attachment. The polypeptide library developed in this work addresses the need for effective biocompatible therapeutics for drug delivery and medical device coatings.     

Combating biofilms

Biofilms are complex microbial communities consisting of microcolonies embedded in a matrix of self-produced polymer substances. Biofilm cells show much greater resistance to environmental challenges including antimicrobial agents than their free-living counterparts. The biofilm mode of life is believed to significantly contribute to successful microbial survival in hostile environments. Conventional treatment, disinfection and cleaning strategies do not proficiently deal with biofilm-related problems, such as persistent infections and contamination of food production facilities. In this review, strategies to control biofilms are discussed, including those of inhibition of microbial attachment, interference of biofilm structure development and differentiation, killing of biofilm cells and induction of biofilm dispersion.     

Tolerance to the antimicrobial peptide colistin in Pseudomonas aeruginosa biofilms is linked to metabolically active cells, and depends on the pmr and mexAB-oprM genes

Bacteria living as biofilm are frequently reported to exhibit inherent tolerance to antimicrobial compounds, and might therefore contribute to the persistence of infections. Antimicrobial peptides are attracting increasing interest as new potential antimicrobial therapeutics; however, little is known about potential mechanisms, which might contribute to resistance or tolerance development towards these compounds in biofilms. Here we provide evidence that a spatially distinct subpopulation of metabolically active cells in Pseudomonas aeruginosa biofilms is able to develop tolerance to the antimicrobial peptide colistin. On the contrary, biofilm cells exhibiting low metabolic activity were killed by colistin. We demonstrate that the subpopulation of metabolically active cells is able to adapt to colistin by inducing a specific adaptation mechanism mediated by the pmr operon, as well as an unspecific adaptation mechanism mediated by the mexAB-oprM genes. Mutants defective in either pmr -mediated lipopolysaccharide modification or in mexAB-oprM -mediated antimicrobial efflux were not able to develop a tolerant subpopulation in biofilms. In contrast to the observed pattern of colistin-mediated killing in biofilms, conventional antimicrobial compounds such as ciprofloxacin and tetracycline were found to specifically kill the subpopulation of metabolically active biofilm cells, whereas the subpopulation exhibiting low metabolic activity survived the treatment. Consequently, targeting the two physiologically distinct subpopulations by combined antimicrobial treatment with either ciprofloxacin and colistin or tetracycline and colistin almost completely eradicated all biofilm cells.