400016,重庆,重庆医科大学附属第一医院急诊医学科
Received:08 November 2019,
Revised:2020-01-03,
Published:15 April 2020
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Xian LIU, Shan XU, Yiming SHEN, et al. Effect of polypeptide bCAT combined with caspofungin on hypha and mature biofilm in
Xian LIU, Shan XU, Yiming SHEN, et al. Effect of polypeptide bCAT combined with caspofungin on hypha and mature biofilm in
目的
2
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
2
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(MBIC
50
),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
2
bCAT及CAS抗成熟生物被膜的MBIC
50
分别为3 105、128 μg/mL,两者联合使用时抗成熟被膜的MBIC
50
分别降至776、4 μg/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(776 μg/mL)、CAS(4 μg/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(776 μg/mL)、CAS(4 μg/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
2
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
2
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
2
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 (MBIC
50
). 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
2
The MBIC
50
after bCAT and CAS treatment was 3 105 and 128 μg/mL
respectively. And the MBIC
50
of bCAT combined with CAS was decreased to 776 μg/mL and 4 μg/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 (776 μg/mL) and CAS (4 μg/mL) groups (
P
<
0.05). The ROS content was (428.96±47.67)%
with significant difference compared with the control
bCAT (776 μg/mL) and CAS (4 μg/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
2
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.
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