酸性氧化电位水对铜绿假单胞菌的杀菌机制研究
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李武平

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R187

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Bactericidal mechanism of electrolyzed oxidizing water against Pseudomonas aeruginosa
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    摘要:

    目的探讨酸性氧化电位水(EOW)对铜绿假单胞菌的杀菌机制。方法以铜绿假单胞菌作为研究对象,从蛋白质渗漏,核酸和细胞膜钙离子通透性等方面对EOW的杀菌机制进行研究,同时以2%戊二醛作为阳性对照组,以生理盐水(NS)作为阴性对照组。结果以EOW、2%戊二醛对铜绿假单胞菌作用30 s杀灭率>99.99%,作用60 s杀灭率可达100.00%。EOW、NS、2%戊二醛作用铜绿假单胞菌60 s后,经BCA法检测蛋白渗漏,分别为(96.00±7.42)、(94.15±7.49)、(216.97±10.35)μg/mL,总体比较,差异有统计学意义(F=613.20,P<0.01),2%戊二醛组高于EOW组、NS组;蛋白渗漏量不随作用时间增加而改变(均P>0.05)。随机多态性扩增DNA电泳图谱显示,EOW组与NS组均在500~1 000 kb间显示亮度较高的密集条带,2%戊二醛组无扩增条带。EOW及2%戊二醛作用后钙离子荧光强度均低于NS组。结论EOW可能的杀菌机制是使铜绿假单胞菌细胞膜通透性受到一定程度的损伤,致使 Ca2+ 泄漏,但未能造成蛋白质泄漏,对其核酸损伤作用不明显,DNA可能并非EOW杀菌靶点。

    Abstract:

    ObjectiveTo investigate the bactericidal mechanism of electrolyzed oxidizing water (EOW) against Pseudomona aeruginosa(P. aeruginosa).MethodsBactericidal mechanism of EOW against P. aeruginosa was studied through intracellular protein leakage, nucleic acid,and cell membrane calcium ion permeability, 2% glutaraldehyde was used as positive control group, and normal saline (NS) was used as negative control group.ResultsThe killing rates of EOW and 2% glutaraldehyde to P. aeruginosa were both>99.99% with 30second contact time, and 100.00% with 60second contact time. After 60second contact with EOW, NS, and 2% glutaraldehyde,the protein leakage of P. aeruginosa detected by bicinchoninic acid (BCA) were (96.00±7.42),(94.15±7.49), and (216.97±10.35)μg/mL, respectively, difference was significant(F=613.20,P<0.01),2% glutaraldehyde group was higher than EOW group and NS group; protein leakage did not change with the increase of contact time(all P>0.05). Electrophoretogram of random amplified polymorphic DNA showed high intensity dense band between 500-1000 Kb in EOW group and NS group, while 2% glutaraldehyde group was without amplified bands. The fluorescence intensity of calcium ion of EOW group and 2% glutaraldehyde group were both lower than that of NS group.ConclusionBactericidal mechanism of EOW may be due to the damage of membrane permeability of P. aeruginosa, which causes Ca2+ leakage, but fails to cause protein leakage, the damage to nucleic acid is not obvious, DNA may not be a bactericidal target of EOW.

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赵凯丽,李武平,张晓娜,等.酸性氧化电位水对铜绿假单胞菌的杀菌机制研究[J]. 中国感染控制杂志,2017,16(1):41-45. DOI:10.3969/j. issn.1671-9638.2017.01.009.
ZHAO Kaili, LI Wuping, ZHANG Xiaona, et al. Bactericidal mechanism of electrolyzed oxidizing water against Pseudomonas aeruginosa[J]. Chin J Infect Control, 2017,16(1):41-45. DOI:10.3969/j. issn.1671-9638.2017.01.009.

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  • 收稿日期:2016-06-01
  • 最后修改日期:2016-08-15
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  • 在线发布日期: 2017-01-22
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