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目的研究三维细胞培养系统(RCCS)模拟微重力对铜绿假单胞菌莫西沙星敏感度的影响。方法采用RCCS建立铜绿假单胞菌ATCC 27853模拟微重力环境培养体系,实验组旋转瓶轴心与地面平行模拟微重力环境,对照组即重力组旋转瓶轴心与地面垂直,分别于培养第1、7、14天取实验组与对照组菌悬液,采用肉汤稀释法检测莫西沙星作用的最低抑菌浓度(MIC)。K-B法检测莫西沙星抑菌环直径,采用荧光染色剂SYTO-9和碘化丙啶(PI)进行染色,检测不同浓度(0.125~64.000μg/ml)莫西沙星作用下实验组与对照组的活菌比例。结果肉汤稀释法检测各时相点实验组与对照组铜绿假单胞菌莫西沙星MIC均为0.25μg/ml。K-B法检测发现,与对照组相比,实验组各时相点莫西沙星抑菌环直径明显延长(P<0.05),且随模拟微重力时相延长,实验组抑菌环直径呈不断延长趋势(P<0.05)。实验组各药物浓度下活菌比例均小于对照组,且随模拟微重力时相延长,同一药物浓度下,实验组活菌比例逐渐减少(P<0.05)。结论 14天RCCS模拟微重力能增加铜绿假单胞菌对莫西沙星的敏感度。
Objective To study the effect of three-dimensional cell culture system (RCCS) simulated microgravity on the sensitivity of Pseudomonas aeruginosa Moxifloxacin. Methods The culture system of Pseudomonas aeruginosa ATCC 27853 simulated microgravity was established by RCCS. The axis of rotation of the rotating bottle in the experimental group was parallel to the simulated microgravity environment. The axis of the rotating bottle in the control group was vertical to the ground. The bacterial suspension of the experimental group and the control group was taken on the 7th and 14th days. The minimum inhibitory concentration (MIC) of the moxifloxacin was determined by broth dilution method. KB method was used to detect the diameter of bacteriostatic ring of moxifloxacin, and stained with fluorescent staining reagent SYTO-9 and propidium iodide (PI) to test the effect of moxifloxacin at different concentrations (0.125 ~ 64.000μg / ml) Of live bacteria ratio. Results Bacillus subtilis Moxifloxacin MIC of 0.25μg / ml in the test group and the control group was detected by broth dilution method at each time point. KB method showed that compared with the control group, the diameter of the antibacterial ring of moxifloxacin in experimental group was significantly prolonged (P <0.05), and the diameter of the antibacterial ring in the experimental group continued to increase as the simulated microgravity phase prolonged Trend (P <0.05). The proportion of viable cells in experimental group was lower than that of control group. With the prolongation of simulated microgravity phase, the proportion of viable cells in experimental group decreased gradually with the same drug concentration (P <0.05). Conclusion The 14-day RCCS simulated microgravity can increase the sensitivity of Pseudomonas aeruginosa to moxifloxacin.