多肽bCAT联合卡泊芬净对白色念珠菌菌丝及成熟生物被膜的作用  

Effect of polypeptide bCAT combined with caspofungin on hypha and mature biofilm in Candida albicans

作　　者：刘贤 许珊[1] 沈一鸣 亢胜男 刘景仑[1] 陈晓迎[1] 张丹[1] LIU Xian;XU Shan;SHEN Yiming;KANG Shengnan;LIU Jinglun;CHEN Xiaoying;ZHANG Dan(Department of Emergency Medicine,the First Affiliated Hospital of Chongqing Medical University,Chongqing,400016,China)

机构地区：[1]重庆医科大学附属第一医院急诊医学科,重庆400016

出　　处：《第三军医大学学报》2020年第7期671-677,共7页Journal of Third Military Medical University

基　　金：重庆市卫计委重点项目(2016ZDXM001);国家自然科学基金面上项目(81071531,81372102)。

摘　　要：目的探讨嗜铬粒蛋白A(chromogranin,CGA)衍生多肽bCAT联合卡泊芬净(caspofungin,CAS)对白色念珠菌菌丝及成熟生物被膜的作用及机制。方法以标准菌株ATCC10231为研究对象,体外建立生物被膜模型,分别用bCAT、CAS、bCAT联合CAS作用于被膜,以RPMI1640干预作为空白对照组(control组)。XTT法检测生物被膜代谢活性的变化,并计算抗成熟生物被膜50%的MIC(MBIC50),RT-PCR检测黏附因子ALS3的mRNA表达,DCFH-DA观察细胞内ROS含量的变化,SYTO9/PI染色观察生物被膜通透性的变化。结果bCAT及CAS抗成熟生物被膜的MBIC50分别为3105、128μg/mL,两者联合使用时抗成熟被膜的MBIC50分别降至776、4μg/mL,RT-PCR检测结果显示bCAT+CAS组处理被膜后ALS3表达量为(1.71±0.65)%,与control、bCAT(776μg/mL)、CAS(4μg/mL)组比较差异具有统计学意义(P<0.05);ROS含量为(428.96±47.67)%,与control、bCAT(776μg/mL)、CAS(4μg/mL)组比较差异具有统计学意义(P<0.05);bCAT+CAS组中死亡菌明显增加,菌丝相真菌明显减少。结论CGA衍生多肽bCAT联合CAS具有协同抗成熟被膜作用,其机制可能与降低黏附因子ALS3的表达而抑制菌丝的形成、增加ROS含量造成氧化损伤相关。Objective To investigate the effect and mechanism of chromogranin(CGA)-derived polypeptide bCAT combined with caspofungin(CAS)on hypha and mature biofilm in Candida albicans(C.albicans).Methods Standard strain ATCC 10231 of C.albicans was employed as the study objective.The changes of biofilm metabolic activity after bCAT and CAS treatment alone or combined together(RPMI1640 as control group)were detected by XTT assay,and the results were calculated for the minimal biofilm inhibitory concentration(MBIC50).The expression of agglutinin-like sequence 3(ALS3)was detected by RT-PCR,and the production of reactive oxygen content(ROS)was observed by DCFH-DA.SYTO9/PI staining was used to detect the fungal membrane permeability.Results The MBIC50 after bCAT and CAS treatment was 3105 and 128μg/mL,respectively.And the MBIC50 of bCAT combined with CAS was decreased to 776μg/mL and 4μg/mL,respectively.The expression of ALS3 in the bCAT+CAS group was(1.71±0.65)%,which had statistical differences from that of the control,bCAT(776μg/mL)and CAS(4μg/mL)groups(P<0.05).The ROS content was(428.96±47.67)%,with significant difference compared with the control,bCAT(776μg/mL)and CAS(4μg/mL)groups(P<0.05).There were more dead C.albicans in mature biofilm in the bCAT+CAS group,and the fungal hypha were decreased significantly.Conclusion CGA-derived polypeptide bCAT combined with CAS has the synergistic anti-mature biofilm effect,and the mechanism may be associated with decreased expression of ALS3 and inhibition of hypha formation,and oxidative damage caused by increased ROS content.

关 键 词：嗜铬粒蛋白A 白色念珠菌 生物被膜 卡泊芬净 协同 

分 类 号：R379.4[医药卫生—病原生物学] R977.6[医药卫生—基础医学]