Objectives.由于抗性的发展,抗真菌剂越来越不有效。此外,由于药物缺乏穿透生物膜的能力,难以治疗形成生物膜的念珠菌生物体。我们正在尝试评估一种新的治疗剂的效果,N-乙酰半胱氨酸(NAC),近平滑念珠菌临床菌株的粘附和生物膜形成。同时,为了检测粘附和生物膜形成相关基因(CpALS6,CpALS7,CpEFG1和CpBCR1)的转录水平变化,此外,探讨药物对生物膜的干扰机制。假设/差距陈述。N-乙酰半胱氨酸(NAC)对来自CRBSIs的近平滑梭菌临床菌株的粘附和生物膜形成具有一定的抑制作用:(1)下调CpEFG1基因的表达,使其成为治疗近扁平梭菌导管相关性血流感染(CRBSIs)的潜在候选药物,(2)调节细胞结构的代谢和生物膜形成因子。方法。为了确定非抗真菌剂是否可以对粘附表现出抑制作用,从念珠菌菌血症患者分离株的总生物膜形成量和代谢活性,将NAC以不同浓度添加到酵母悬浮液中,分别。逆转录检测BCR1敲除菌株中粘附相关基因(CpALS6和CpALS7)和生物膜形成相关因子(CpEFG1和CpBCR1)的转录水平,CP7和CP5临床菌株中存在NAC。为进一步探讨NAC对近融合梭菌生物膜的作用机制,RNA测序用于计算基因表达,比较样本之间的差异。基因本体论(GO)富集分析有助于说明两个特定样品在功能水平上的差异。结果。高浓度的NAC减少近扁平梭菌中生物膜形成的总量。与NAC共同孵育后,CP7和CP5临床菌株中CpEFG1的表达均降低,与未处理的菌株相比,CpBCR1的转录水平没有显着变化。GO富集分析表明,NAC干预后,细胞结构的代谢和生物膜形成因子均受到调控。Conclusions.非抗真菌剂NAC通过下调CpEFG1基因的表达对临床分离株生物膜形成具有一定的抑制作用,使其成为治疗近扁平梭菌导管相关性血流感染的潜在候选药物。
Objectives. Anti-fungal agents are increasingly becoming less effective due to the development of resistance. In addition, it is difficult to treat Candida organisms that form
biofilms due to a lack of ability of drugs to penetrate the
biofilms. We are attempting to assess the effect of a new therapeutic agent, N-acetylcysteine (NAC), on adhesion and biofilm formation in Candida parapsilosis clinical strains. Meanwhile, to detect the transcription level changes of adhesion and biofilm formation-associated genes (CpALS6, CpALS7, CpEFG1 and CpBCR1) when administrated with NAC in C. parapsilosis strains, furthermore, to explore the mechanism of drug interference on biofilms.Hypothesis/Gap statement. N-acetylcysteine (NAC) exhibits certain inhibitory effects on adhesion and biofilm formation in C. parapsilosis clinical strains from CRBSIs through: (1) down-regulating the expression of the CpEFG1 gene, making it a highly potential candidate for the treatment of C. parapsilosis catheter-related bloodstream infections (CRBSIs), (2) regulating the metabolism and biofilm -forming factors of cell structure.Methods. To determine whether non-antifungal agents can exhibit inhibitory effects on adhesion, amounts of total biofilm formation and metabolic activities of C. parapsilosis isolates from candidemia patients, NAC was added to the yeast suspensions at different concentrations, respectively. Reverse transcription was used to detect the transcriptional levels of adhesion-related genes (CpALS6 and CpALS7) and biofilm formation-related factors (CpEFG1 and CpBCR1) in the BCR1 knockout strain, CP7 and CP5 clinical strains in the presence of NAC. To further explore the mechanism of NAC on the
biofilms of C. parapsilosis, RNA sequencing was used to calculate gene expression, comparing the differences among samples. Gene Ontology (GO) enrichment analysis helps to illustrate the difference between two particular samples on functional levels.Results. A high concentration of NAC reduces the total amount of biofilm formation in C. parapsilosis. Following co-incubation with NAC, the expression of CpEFG1 in both CP7 and CP5 clinical strains decreased, while there were no significant changes in the transcriptional levels of CpBCR1 compared with the untreated strain. GO enrichment analysis showed that the metabolism and biofilm-forming factors of cell structure were all regulated after NAC intervention.Conclusions. The non-antifungal agent NAC exhibits certain inhibitory effects on clinical isolate biofilm formation by down-regulating the expression of the CpEFG1 gene, making it a highly potential candidate for the treatment of C. parapsilosis catheter-related bloodstream infections.