厚朴煎剂对白色念珠菌及其生物膜影响的初步研究

目的 观察厚朴煎剂对白色念珠菌(Candida albicans)芽管形成、早期黏附基因及成熟期生物膜的影响。方法 采用芽管形成试验检测厚朴煎剂对白色念珠菌芽管形成的影响;实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)检测厚朴煎剂作用后,凝集素样序列1(agglutinin-like sequencefamily1,ALS1)、ALS2、ALS3基因的表达情况;荧光显微镜及扫描电镜观察厚朴煎剂对白色念珠菌成熟期生物膜的影响;共聚焦显微镜观察厚朴煎剂对生物膜中菌活性的影响。结果 芽管形成试验结果显示,厚朴煎剂可抑制白色念珠菌的芽管形成且抑制作用具有浓度依赖性(P＜0.05)。厚朴煎剂能使ALS1、ALS2、ALS3基因表达下调且具有浓度依赖性(P＜0.05)。荧光显微镜下空白对照组生物膜以菌丝为主,厚朴煎剂作用24、48及72 h后生物膜结构被破坏。扫描电镜结果也提示厚朴煎剂可以破坏生物膜的结构。共聚焦显微镜下空白对照组白色念珠菌以活菌为主,15.625 mg/L厚朴煎剂作用48 h后死菌数量明显增加。结论 厚朴煎剂可通过下调ALS1、ALS2、ALS3基因抑制白色念珠菌早期黏附,同时可通过破坏成熟期生物膜的结构促进药物进入生物膜内抑制芽管形成和发挥杀菌作用。

Effect of Magnolia officinalis decoction on Candida albicans and its biofilm

Objective To observe the effects of Magnolia officinalis decoction on the formation of germ tube, the early adhesion gene and mature biofilm of Candida albicans(C.albicans). Methods Bud tube formation test was used to detect the effect of Magnolia officinalis decoction on the formation of bud tube C. albicans. The expression of agglutinin-like sequencefamily (ALS1), ALS2 and ALS3 genes was detected in real-time using fluorescence quantitative real-time PCR (qRT PCR). The effects of Magnolia officinalis decoction on the biofilm of C. albicansat maturity were observed using fluorescence microscope and scanning electron microscope(SEM). The effect of Magnolia officinalis decoction on bacterial activity in biofilm was observed by confocal microscope. Results The results obtained from the formation of germ tube test showed that Magnolia officinalis decoction could inhibit the formation of germ tube C.albicans in a concentration dependent manner (P<0.05). Magnolia officinalis decoction could reduce the expression of ALS1, ALS2 and ALS3 genes in a concentration dependent manner (P<0.05).Under fluorescence microscope, the biofilm in the blank control group was mainly mycelium. The biofilm structure was destroyed after the Magnolia officinalis decoction for 24, 48 and 72. SEM results also showed that Magnolia officinalis decoction could result in the destruction of the biofilm structure. Confocal microscopy showed that C. albicans in the blank control group was mainly living bacteria, and the number of dead bacteria increased significantly after 48 hours of Magnolia officinalis decoction at 15.625 mg/L. Conclusion The Magnolia officinalis decoction can inhibit the early adhension of C. albicans by down regulating ALS1, ALS2 and ALS3 genes.At the same time,it can promote the entry of drugs into biofilm by destroying the structure of mature biofilm in inhibit the formation of germ tube and play a bactericidal role.