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目的:采用激光共聚焦显微镜(CLSM)技术考察穿心莲内酯对铜绿假单胞菌PAO1生物被膜形成的影响,进一步揭示穿心莲内酯的抗菌机制。方法:使用玻璃爬片生物被膜培养法,在不同浓度穿心莲内酯(10μg/ml、1μg/ml、0.1μg/ml和0.01μg/ml)条件下孵育PAO1,并分别于24h、48h及72h取出玻璃爬片,用硫氰酸荧光素标记的刀豆蛋白和碘化吡啶双重免疫荧光染色,采用激光共聚焦显微镜技术观察穿心莲内酯对铜绿假单胞菌生物被膜形成的影响,并测量生物被膜的厚度。结果:断层图像显示和空白对照组比较,用药后PAO1生物被膜生长不完全,出现多种形态,其中在0.1μg/ml剂量下,生物被膜呈破碎絮状;与空白对照组比较,穿心莲内酯在0.1μg/ml、0.01μg/ml剂量下,可明显影响PAO1生物被膜的生长厚度。结论:穿心莲内酯可能具有干扰铜绿假单胞菌PAO1生物被膜形成的作用。
OBJECTIVE: To investigate the effect of andrographolide on the biofilm formation of Pseudomonas aeruginosa PAO1 by using confocal laser scanning microscopy (CLSM), further revealing the antibacterial mechanism of andrographolide. Methods: PAO1 was incubated under different concentrations of andrographolide (10μg / ml, 1μg / ml, 0.1μg / ml and 0.01μg / ml) using glass slide biofilm culture method and was removed at 24h, 48h and 72h respectively Glass slides, double-immunofluorescent staining with concanavalin labeled with fluorescein thiocyanate and pyridine iodide were used to observe the effect of andrographolide on the biofilm formation of Pseudomonas aeruginosa by laser confocal microscopy. thickness of. Results: Compared with the blank control group, the PAO1 biofilm did not grow completely and showed many morphological changes. The biofilm was broken and flocculent at the dosage of 0.1μg / ml. Compared with the blank control group, the andrographolide In 0.1μg / ml, 0.01μg / ml dose, can significantly affect the growth of PAO1 biofilm thickness. Conclusion: Andrographolide may interfere with the formation of PAO1 biofilm in Pseudomonas aeruginosa.