|
|
|
| Effect of RNAⅢ inhibitor peptide on the expression of virulence factors of Staphylococcus aureus at different growth phases |
| ZHOU Ying1 HOU Zheng2 XUE Xiaoyan2 WEI Ming1 LI Fen1 YAO Jia3 ZHANG Ye3 WANG Yang3 |
1.Department of Pharmacology, Xi’an Medical University, Shaanxi Province, Xi’an 710021, China;
2.Department of Pharmacology, Air Force Medical University, Shaanxi Province, Xi’an 710032, China;
3.Molecular Virology and Viral Immunology Laboratory, Xi’an Medical University, Shaanxi Province, Xi’an 710021, China |
|
|
|
Abstract Objective To explore the role of RNAⅢ inhibitory peptide (RIP) in regulating the expression of staphylolysin and adhesion-related proteins at different growth phases. Methods The methicillin-resistant Staphylococcus aureus was inoculated to medium proportionally, and the bacteria cultured overnight were transferred to TSB medium (containing 0.5% glucose), respectively, and the concentration was adjusted to 1×108 CFU/mL. The RIP group was added with RIP derivatives (RIP1183) designed and synthesized by research group, while the control group was added with sterile phosphate buffer. Its effects on the growth curve and biofilm formation were examined. Quantitative real-time fluorescence polymerase chain reaction was used to detect the expression levels of RNAⅢ, human leucocyte antigen (HLA), icaA, and fnbA during the bacterial retardation phase (2 h), logarithmic phase (6 h), and stable phase (10 h), and compared with the control group. Results Compared with the control group, RIP1183 did not affect the growth of bacteria, and the difference was not statistically significant (P ＞ 0.05). Compared with the control group, the formation of bacterial biofilms in the RIP group was significantly reduced (P ＜ 0.01). In the bacterial retardation phase (2 h), the expression of RNAⅢ in RIP group was obviously decreased (P < 0.05). In the logarithmic phase (6 h), compared with the control group, the expression of RNAⅢ and HLA were obviously decreased (P ＜ 0.01). In the stable phase (10 h), the expression of icaA, fnbA and RNAⅢ were obviously decreased (P ＜ 0.05 or P ＜ 0.01). Conclusion RIP1183 inhibits the expression of RNAⅢ, an effector molecule of the accessory gene regulation system, and significantly reduces the expression of different virulence factors in different phases, thereby destroying the mechanism of bacteria’s adaptation to the environment, which may be one of the mechanisms of its antibacterial activity and anti-biofilm formation.
|
|
|
|
|
|
[1] ?譧lvarez A，Fernández L，Gutiérrez D，et al. Methicillin-Resistant Staphylococcus aureus in Hospitals：Latest Trends and Treatments Based on Bacteriophages [J]. J Clin Microbiol，2019，57（12）：e01006-19.
[2] Mitsutake K，Niwaya K. Infections associated with surgical implants [J]. N Engl J Med，2004，351（2）：193-195.
[3] Kot B，Sytykiewicz H，Sprawka I. Expression of the Biofilm-Associated Genes in Methicillin-Resistant Staphylococcus aureus in Biofilm and Planktonic Conditions [J]. Int J Mol Sci，2018，19（11）：3487.
[4] Tran VG，Venkatasubramaniam A，Adhikari RP，et al. Efficacy of Active Immunization with Attenuated Alpha-Hemolysin and Panton-Valentine Leukocidin in a Rabbit Model of Staphylococcus aureus Necrotizing Pneumonia [J]. J Infect Dis，2020，221（2）：267-275.
[5] Ma B，Zhou Y，Li M，et al. RIP-V improves murine survival in a sepsis model by down-regulating RNAIII expression and α-hemolysin release of methicillin-resistant Staphylococcus aureus [J]. Pharmazie，2015，70（2）：81-87.
[6] Zhou Y，Niu C，Ma B，et al. Inhibiting PSMα-induced neutrophil necroptosis protects mice with MRSA pneumonia by blocking the agr system [J]. Cell Death Dis，2018，9（3）：362.
[7] Boles BR，Horswill AR. Agr-mediated dispersal of Staphylococcus aureus biofilms [J]. PLoS Pathog，2008，4（4）：e1000052.
[8] Zhou Y，Ma B，Gao H，et al. An oligomerization of RNAⅢ inhibiting peptide Inhibits Adherence and Biofilm Formation of Methicillin-Resistant Staphylococcus aureus in vitro and in vivo [J]. Microb Drug Resisit，2016，22（3）：193-201.
[9] Balaban N，Cirioni O，Giacometti A，et al. Treatment of Staphylococcus aureus biofilm infection by the quorum-sensing inhibitor RIP [J]. Antimicrob Agents Chemother，2007，51（6）：2226-2229.
[10] Bhardwaj AK，Vinothkumar K，Rajpara N. Bacterial quorum sensing inhibitors：attractive alternatives for control of infectious pathogens showing multiple drug resistance [J]. Recent Pat Antiinfect Drug Discov，2013，8（1）：68-83.
[11] Garrigós C，Murillo O，Lora-Tamayo J，et al. RNAⅢ inhibiting peptide against foreign-body infection by methicillin-resistant Staphylococcus aureus [J]. J Infect，2012， 65（6）：586-588.
[12] Zhao J，Chen M，Quan CS，et al. Mechanisms of quorum sensing and strategies for quorum sensing disruption in aquaculture pathogens [J]. J Fish Dis，2015，38（9）：771-786.
[13] Tang F，Li L，Meng XM，et al. Inhibition of alpha-hemolysin expression by resveratrol attenuates Staphylococcus aureus virulence [J]. Microb Pathog，2019，127：85-90.
[14] Romp E，Arakandy V，Fischer J，et al. Exotoxins from Staphylococcus aureus activate 5-lipoxygenase and induce leukotriene biosynthesis [J]. Cell Mol Life Sci，2019：1-18.
[15] Bimanand L，Taherikalani M，Jalilian FA，et al. Association between biofilm production，adhesion genes and drugs resistance in different SCCmec types of methicillin resistant Staphylococcus aureus strains isolated from several major hospitals of Iran [J]. Iran J Basic Med Sci，2018，21（4）：400-403.
[16] Azara E，Longheu C，Sanna G，et al. Biofilm formation and virulence factor analysis of Staphylococcus aureus isolates collected from ovine mastitis [J]. J Appl Microbiol，2017，123（2）：372-379.
[17] Yousefi M，Pourmand MR，Fallah F，et al. A Characterization of Staphylococcus aureus Biofilm Formation in Urinary Tract Infection [J]. Iran J Public Health，2016， 45（4）：485-493.
[18] Lade H，Park JH，Chung SH，et al. Biofilm Formation by Staphylococcus aureus Clinical Isolates is Differentially Affected by Glucose and Sodium Chloride Supplemented Culture Media [J]. J Clin Med，2019，8（11）：1853.
[19] Brackman G，Coenye T. Inhibition of Quorum Sensing in Staphylococcus spp [J]. Curr Pharm Des，2015，21（16）：2101-2108.
[20] Donlan RM，Costerton JW. Biofilms：survival mechanisms of clinically relevant microorganisms [J]. Clin Microbiol Rev，2002，15（2）：167-193.
[21] Bhattacharya M，Berends ETM，Chan R，et al. Staphylococcus aureus biofilms release leukocidins to elicit extracellular trap formation and evade neutrophil-mediated killing [J]. Proc Natl Acad Sci U S A，2018，115（28）：7416-7421.
[22] Brackman G，Coenye T. Quorum sensing inhibitors as anti-biofilm agents [J]. Curr Pharm Des，2015，21（1）：5-11. |
|
|
|
