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目的探讨木犀草素对粪肠球菌生物被膜的抑制效果,并进一步研究木犀草素对粪肠球菌毒力因子转录表达水平的影响。方法建立粪肠球菌的体外生物被膜模型,5组实验组分别加入浓度为0.5、1、2、4和8mg/mL的木犀草素,同步培养24h后,通过MTT检测各组的抑菌率。利用激光共聚焦显微镜观察不同浓度的木犀草素作用24h后的抑菌效果。最后选取适宜的浓度(2、4、8mg/mL)的木犀草素与粪肠球菌共同培养24h后,通过RT-PCR观察各组粪肠球菌毒力因子gelE、esp、ebpA的转录表达水平。结果 MTT和CLSM结果显示:随着浓度的增加,木犀草素对粪肠球菌生物被膜的抑制效果越来越好(P<0.05)。其中以8mg/mL组的抑菌效果最好;浓度为2、4、8mg/mL的木犀草素对其毒力因子gelE、esp、ebpA的mRNA的表达均有较好的抑制作用,并且随着药物浓度的增加木犀草素对其毒力因子的抑制效果也越来越好(P<0.05)。当药物浓度为8mg/mL时,可完全抑制gelE、esp、ebpA的mRNA的转录表达。结论木犀草素对粪肠球菌生物被膜有抑制作用,与此同时还能不同程度的抑制其毒力因子gelE、esp、ebpA的转录表达水平。
Objective To investigate the inhibitory effect of luteolin on the biofilm of Enterococcus faecalis and further study the influence of luteolin on the transcriptional expression of virulence factors of Enterococcus faecalis. Methods The biofilm model of Enterococcus faecalis was established in vitro. Luteolin, 0.5, 1, 2, 4 and 8 mg / mL respectively, was added to the experimental groups of 5 groups. After 24h incubation, MTT assay was used to detect the inhibitory rate of each group. The effect of different concentrations of luteolin on the bacteriostasis was observed by laser scanning confocal microscopy. Finally, the appropriate concentration (2,4,8mg / mL) of luteolin with Enterococcus faecalis co-cultured 24h, RT-PCR observation of each group Enterococcus faecal virulence factors gelE, esp, ebpA transcriptional expression levels. Results The results of MTT and CLSM showed that with the increase of concentration, the inhibitory effect of luteolin on Enterococcus faecalis biofilm was better (P <0.05). Among them, the antibacterial effect of 8mg / mL group was the best; luteolin at 2,4,8mg / mL had a good inhibitory effect on the mRNA expression of gelE, esp and ebpA, With the increase of drug concentration, luteolin also inhibited its virulence factors better and better (P <0.05). When the drug concentration was 8mg / mL, the transcriptional expression of gelE, esp and ebpA mRNAs was completely inhibited. Conclusion Luteolin could inhibit the biofilm formation of Enterococcus faecalis and inhibit the transcriptional expression of gelE, esp and ebpA to some extent.