群体感应分子Farnesol联合CSA对白色念珠菌生物膜作用的实验研究

The effect of the quorum sensing molecule farnesol combined with CSA on Candida albicans biofilm

目的探讨群体感应分子法尼醇(Farnesol)联合钙调神经磷酸酶抑制剂环孢菌素A(CSA)对白色念珠菌的生物膜形成、形态转换、生长及活性、耐药性、凋亡反应和氧化应激反应的作用及其相关机制。方法体外构建生物膜状态白色念珠菌,分别经CSA、Farnesol、Farnesol联合CSA处理后,用qRT-PCR法检测生物膜状态白色念珠菌中生物膜形成过程中的相关基因CDR1、CEK1、CPH1、EFG1、ERG11、GPR1、HAT1、MDR1、HWP1、ALS3、TUP1mRNA表达水平的变化。结果白色念珠菌均具有不同程度的产膜能力,且单用CSA、Farnesol抗真菌药物对白色念珠菌生物膜的抑制率很低,而联合使用时,受试菌生物膜生长状态受到明显抑制。从临床血流感染中分离获得1株强产生物被膜白色念珠菌,命名为CA NX05;经不同药物组处理后生物膜状态白色念珠菌中的生物膜菌丝形成相关基因CEK1、CPH1、EFG1、ERG11、GPR1、HAT1、HWP1、ALS3、TUP1与其耐药性相关基因CDR1、MDR1的mRNA表达量与空白处理组相比均显著降低(F_(CEK1)=15.725,F_(CPH1)=6.230,F_(EFG1)=87.817,F_(ERG11)=17.527,F_(GPR1)=21.793,F_(HAT1)=65.318,F_(HWP1)=550.531,F_(ALS3)=10.514,F_(TUP1)=16.580,F_(CDR1)=8.089,F_(MDR1)=7.228;F>P,P<0.05)。结论群体感应分子Farnesol联合CSA处理较CSA及Farnesol处理能更有效地抑制白色念珠菌生物膜的形成,从而使其对抗真菌药物的敏感性增加,为临床合理使用抗真菌药物提供了科学导向。

Objective To research the effects and mechanisms of the quorum sensing molecule farnesol combined with CSA on the biological characteristics of C.albicans,such as its formation of biofilm,phenotype switching,growth and proliferation,drug resistance,apoptotic response,and oxidative stress response. Methods Candida albicans biofilm was formed in vitro and then treated with CSA,farnesol,or a combination with the two drugs,after which changes in mRNA of genes related to biofilm formation such as CDR1,CEK1,CPH1,EFG1,ERG11,GPR1,HAT1,MDR1,HWP1,ALS3,and TUP1 were detected with qRT-PCR. Results CSA alone and farnesol alone had little impact on inhibiting biofilm formation and inhibiting different concentrations of C.albicans,but these two drugs had a marked inhibitory effect when combined.A strain of C.albicans that formed a strong biofilm was isolated from bloodstream infection samples and that strain was designated CA NX05.After the biofilm-forming C.albicans was treated with different drugs,there was a marked decrease in expression of mRNA of genes related to biofilm formation such as CDR1,CEK1,CPH1,EFG1,ERG11,GPR1,HAT1,MDR1,HWP1,ALS3,and TUP1 and mRNA of genes related to drug resistance such as CDR1 and MDR1（FCEK1=15.725,FCPH1=6.230,FEFG1=87.817,FERG11=17.527,FGPR1=21.793,FHAT1=65.318,FHWP1=550.531,FALS3=10.514,FTUP1=16.580,FCDR1=8.089,and FMDR1=7.228;F〉P）. Conclusion The quorum sensing molecule farnesol combined with CSA more efficiently inhibited the ability of C.albicans to form a biofilm compared to other drugs,increasing the sensitivity of C.albicans to antifungals.This research has enabled more rational use of antifungals.