蒋治楠 王涛 汪滢 于勇 毕良佳 马巍.低频超声对金黄色葡萄球菌生物膜的促渗作用研究[J].,2016,16(14):2618-2623 |
低频超声对金黄色葡萄球菌生物膜的促渗作用研究 |
Effect of Low-Frequency Ultrasound on Permeation of Staphylococcus Aureus Biofilm* |
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DOI: |
中文关键词: 低频超声 金黄色葡萄球菌 生物膜 促渗 荧光光谱 |
英文关键词: Low frequency ultrasound Staphylococcus aureus Biological membrane Permeation Fluorescence spectrum |
基金项目:高等学校博士学科点专项科研基金联合资助课题(20132307110018);黑龙江省青年科学基金项目(QC2016119) |
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中文摘要: |
目的:探讨低频超声对血卟啉单甲醚(Hematoporphyrin monomerthyl ether, HMME)在金黄色葡萄球菌生物膜中渗透效果的
影响。方法:将直径25 mm,孔径0.22 滋m 的微孔滤膜平铺于LB 琼脂培养基表面,再将纯种的金黄色葡萄球菌(ATCC6538)菌液,
通过均匀涂布接种到滤膜表面,37 ℃ 恒温培养48 h 获得实验所需的生物膜模型。将培养所得生物膜样本随机分为空白对照组,
单独超声组,单独药物组和实验组,其中实验组按超声强度(0.5 w/cm2或1 w/cm2)和超声作用时间(1 min、2 min 或3 min)不同又
分为实验(A、B、C、D、E、F)组。所用渗透药物为20 ug/mL的HMME 溶液。对照组使用等量的生理盐水孵育,孵育时间为3 min,
单独超声组也用生理盐水孵育并加0.5 W/cm2的超声作用3 min。HMME 在405 mm激发光下有特征性光谱,通过测量光谱曲线
下积分面积,可间接反映生物膜中HMME 渗入量。结果:实验A组(0.5 w/cm2,1 min)的荧光光谱曲线下积分面积显著高于单独
药物组(P<0.05)。声强相同时,随作用时间增加,所得光谱曲线下积分面积显著增加(P<0.01 或P<0.05);作用时间相同时,声强越
大,光谱曲线下积分面积亦越大(P<0.01)。结论:低频超声可以显著增加HMME渗透进入金黄色葡萄球菌生物膜的量,且渗透效
果与超声作用时间和超声声强正相关。 |
英文摘要: |
Objective:To investigate the effect of low-frequency ultrasound on permeation of Hematoporphyrin monomerthyl
ether (HMME) in Staphyloccus aureus biofilm.Methods:Flatting microporous membrane (Φ 6 mm) on LB agar surface,
bacteria (ATCC6538) were inoculated onto the surface of membrane. To obtain the required biofilm model, the
bacteria was cultured for 48 h under the condition of constant temperature(37 ℃). The cultivated biofilm samples were randomly divided
into 4 groups, control group, single ultrasound group, single drug group and the experimental group (ultrasound + drug group).
Furthermore, the experimental group was subdivided into different experiment (A, B, C, D, E, F) groups based on the different ultrasonic
intensity (0.5 w/cm2 or 1 w/cm2) and ultrasonic action time (1 min, 2 min or 3 min). The medications used were 20 ug/mL HMME
solution. The same amount of normal saline was used in the control group and incubated for 3 min. The individual ultrasonic group was
incubated with normal saline and added 0.5 w/cm2 ultrasonic treatment for 3 min. We tested the characteristic spectrum of HMME was
tested which was excited by 405 nm laser in each sample, then measured the integral size of area of each spectrum under the curve and
compared the size of the areas which would indirectly reflect the amount of permeated HMME.Results:The area of fluorescence integral
under the curve in ultrasound + drug group was significantly higher than that of single drug group (P<0.05).When the ultrasonic intensity
was same, the integral size of area under the curve was increased significantly with the increase of reaction time(P<0.01 or P<0.05).
When the time was same, the ultrasonic intensity was greater, then the integral area under the curve was bigger (P<0.01).Conclusion:Low frequency ultrasound can significantly increase the quantity of HMME which penetrates into the Staphylococcus aureus biofilm. The
effect of permeation is positively correlated with both the ultrasonic intensity and the time of ultrasonic treatment. |
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