[1]胡建娥,夏云.肽核酸体外抑制铜绿假单胞菌PAO1生物膜形成的研究[J].陆军军医大学学报(原第三军医大学学报),2011,33(03):294-298.
　Hu Jiane,Xia Yun.Inhibitory effect of peptide nucleic acid on biofilm formation of Pseudomonas aeruginosa PAO1 in vitro[J].J Amry Med Univ (J Third Mil Med Univ),2011,33(03):294-298.
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肽核酸体外抑制铜绿假单胞菌PAO1生物膜形成的研究(/HTML )
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陆军军医大学学报（原第三军医大学学报）[ISSN:1000-5404/CN:51-1095/R]

卷:
33卷
期数:
2011年第03期
页码:
294-298
栏目:
论著
出版日期:
2011-02-15

文章信息/Info

Title:
Inhibitory effect of peptide nucleic acid on biofilm formation of Pseudomonas aeruginosa PAO1 in vitro
作者:
胡建娥; 夏云
重庆医科大学附属第一医院检验科
Author(s):
Hu Jian’e;  Xia Yun
Department of Laboratory Medicine, First Affiliated Hospital , Chongqing Medical University, Chongqing, 400016, China
关键词:
肽核酸; 铜绿假单胞菌; 生物膜
Keywords:
PNA;  Pseudomonas aeruginosa;  biofilm
分类号:
R372; R378.991; R394.3
文献标志码:
A
摘要:
目的   研究肽核酸（peptide nucleic acid，PNA）对铜绿假单胞菌生物膜形成的抑制作用，探讨其潜在的抗铜绿假单胞菌生物膜形成的应用价值。   方法     采用计算机软件设计和斑点杂交联合筛选出能与铜绿假单胞菌PAO1早期生物膜形成关键基因motA mRNA结合紧密的反义寡核苷酸序列，根据其序列合成肽核酸（PNA）并连接穿膜肽(cell penetrating peptide ,CCPs)（KFF）3K形成CCPs-PNA，分别采用不同浓度的CCPs、PNA和CCPs-PNA处理铜绿假单胞菌PAO1，定量测定并用显微镜观察其对生物膜形成的抑制作用。   结果     PNA和CCPs-PNA对铜绿假单胞菌PAO1生物膜的早期形成都有抑制作用，并且随着浓度的增加，其抑制作用也增加，但CCPs-PNA对生物膜形成的抑制明显优于PNA组。1、5、10 μmol/L的（KFF）3K-PNA对PAO1生物膜形成的抑制率分别约为30%、50%、70%。而1 μmol/L的PNA对PAO1生物膜形成基本没有影响，5、10 μmol/L的PNA对PAO1生物膜形成的抑制率却分别为3%和10%左右。各个浓度的CCPs对细菌生物膜的形成都没有影响。与对照组相比，CCPs-PNA处理之后的PAO1，其运动能力明显下降，PAO1处理组与对照组在运动培养基上的扩散直径之比约为3∶5。   结论     针对motA基因的PNA通过抑制PAO1的运动能力、降低吸附，从而对铜绿假单胞菌PA01起始阶段的生物膜形成产生抑制作用，穿膜肽（KFF）3K大大增强了此作用。推测铜绿假单胞菌的motA基因或其编码产物可能是一个良好的抗铜绿假单胞菌生物膜形成的靶点。
Abstract:
Objective    To study the inhibitory effect of peptide nucleic acid (PNA) on biofilm formation of Pseudomonas aeruginosa PAO1 in vitro.     Methods   Sequences of antisense oligonucleotide binding closely to the key gene motA for the early biofilm formation of Pseudomonas aeruginosa PAO1 were designed using Mfold software and dot hybridization. PNA was synthesized according to the sequences of antisense oligonucleotide which was linked to the cell penetrating peptide (CCPs) (KFF)3K to form CCPs-PNA. Pseudomonas aeruginosa PAO1 was treated with CCPs, PNA and CCPs-PNA at different concentrations, and the inhibitory effect of PNA on biofilm formation of Pseudomonas aeruginosa PAO1 was quantitatively detected and observed under a microscope.    Results   PNA and CCPs-PNA inhibited the biofilm formation of Pseudomonas aeruginosa PAO1 in a dose-dependent manner. The inhibitory effect of CCPs-PNA was better than that of PNA on biofilm formation of Pseudomonas aeruginosa PAO1. The inhibitory rate of (KFF)3K-PNA was 30%, 50% and 70%, respectively, for the biofilm formation of Pseudomonas aeruginosa PAO1 at the concentration of 1, 5, and 10 μmol/L. The inhibitory rate of PNA was 3% and 10%, respectively, for the biofilm formation of Pseudomonas aeruginosa PAO1 at the concentration of 5 μmol/L and 10 μmol/L. No effect of CCPs at different concentration was observed on the biofilm formation of Pseudomonas aeruginosa PAO1. The motility of Pseudomonas aeruginosa PAO1 was significantly decreased after treated with CCPs-PNA. The PAO1 radial diameter ratio was 3∶5 between (KFF)3K-PNA treatment group and control group.    Conclusion   The PNA targeting motA gene can inhibit the biofilm formation of Pseudomonas aeruginosa PAO1 at the initial stage by suppressing its motility and adsorption, which can be greatly increased by (KFF)3K, suggesting that the motA gene or its encoded product can be used a good target to prevent biofilm formation of Pseudomonas aeruginosa PAO1.

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更新日期/Last Update: 2011-01-31