论文部分内容阅读
目的利用激光共聚焦显微镜(Confocal Laser Scanning Microscopy,CLSM)观察大蒜素对铜绿假单胞菌PAO1菌株生物膜(biofilms,BF)形成过程的影响,并用ISA(Image Structure Analyzer)软件对BF结构进行定量化分析。方法体外建立1 d、3 d和7 d组PAO1菌株BF模型,每组分3小组,分别用生理盐水、大蒜素128μg/mL、大蒜素10μg/mL作用6 h。通过荧光探针SYTO9/PI标记,利用CLSM进行动态观察,并用ISA软件对其BF结构进行定量分析。结果 (1)生理盐水对照组铜绿假单胞菌生物膜厚度随时间的增加而增加,但在后3 d增加的速度减慢;同时区域孔率(Areal Porosity,AP)呈下降趋势,平均扩散距离(Average Diffusion Distance,ADD)有逐渐增加趋势,结构熵(Textural Entropy,TE)有增加的趋势。(2)7 d组模型中,大蒜素128μg/mL作用后,BF厚度由(28.83±0.67)μm减低到(16.50±0.69)μm;AP由0.76±0.10增加到0.92±0.02;TE由6.78±0.93减低到5.61±0.55。10μg/mL大蒜素干预后,有同样趋势,但效应不如高浓度明显。(3)1 d、3 d模型组,大蒜素作用后同对照组相比,厚度、AP、TE的差异均有统计学意义,趋势同7 d组。结论通过ISA软件对干预前后的BF进行结构定量分析,大蒜素对各个时间段BF的形成均有影响,高浓度效果更显著。
Objective To investigate the effect of allicin on the formation of biofilms (BF) by using the confocal laser scanning microscopy (CLSM) Analysis. Methods The model of PAO1 strain BF was established in vitro on 1 d, 3 d and 7 d groups, and each group consisted of 3 subgroups. The rats were treated with saline, allicin 128 μg / mL and allicin 10 μg / mL for 6 h. Fluorescence probe SYTO9 / PI labeling, the use of CLSM dynamic observation, and its use of ISA software to BF structure quantitative analysis. Results (1) The thickness of biofilm of Pseudomonas aeruginosa increased with the increase of time in physiological saline control group, but decreased slowly at the third day, meanwhile the regional porosity (Areal Porosity, AP) Average Diffusion Distance (ADD) tends to increase gradually, while Textural Entropy (TE) tends to increase. (2) The BF thickness decreased from (28.83 ± 0.67) μm to (16.50 ± 0.69) μm and the AP increased from 0.76 ± 0.10 to 0.92 ± 0.02 after 128 μg / mL allicin treatment in 7-day model group. 0.93 reduced to 5.61 ± 0.55.10μg / mL allicin intervention, the same trend, but the effect is not as obvious as the high concentration. (3) Compared with the control group, the differences in thickness, AP, TE in model group and allicin after 1 d, 3 d group were all statistically significant, with the trend being the same as that in 7 d group. Conclusions The structure and quantitative analysis of BF before and after intervention by ISA software were carried out. Allicin had an impact on the formation of BF in various time periods, and the effect of high concentration was more significant.