Ding Xiurong, Liu Zhizhong, Su Jianrong, Yan Donghui
Clinical Laboratory Center, Beijing Friendship Hospital, Capital Medical University, Beijing 100050, China.
BMC Microbiol. 2014 Mar 28;14:80. doi: 10.1186/1471-2180-14-80.
Candida albicans can form biofilms on intravenous catheters; this process plays a key role in the pathogenesis of catheter infections. This study evaluated the effect of human serum (HS) on C. albicans biofilm formation and the expression of adhesion-related genes in vitro. A C. albicans laboratory strain (ATCC90028) and three clinical strains were grown for 24 h in RPMI 1640 supplemented with HS or RPMI 1640 alone (as a control). The growth of biofilm cells of four strains was monitored by a Live Cell Movie Analyzer, and by XTT reduction assay. The expression of the adhesion-related genes BCR1, ALS1, ALS3, HWP1 and ECE1 was analyzed by RT-PCR at three time points (60 min, 90 min, and 24 h).
In the adhesion phase, C. albicans cells kept a Brownian movement in RPMI medium containing HS until a large number of germ tubes were formed. In the control group, C. albicans cells quickly adhered to the bottom of the reaction plate. Compared with RPMI 1640, medium supplemented with 3-50% HS caused a significant decrease in biofilm development (all p < 0.001). However, the presence of HS had no significant inhibitory effect on the pre-adhered biofilms (all p > 0.05). Biofilm formation was also inhibited by heat-inactivated and proteinase K pre-treated HS. The presence of 50% HS did not significantly affect the planktonic growth of C. albicans (p > 0.05). At three time points, HS inhibited expression of the ALS1 and ALS3 genes and promoted expression of the HWP1 and ECE1 genes. Significant up-regulation of BCR1 was observed only at the 90-min point.
Human serum reduces biofilm formation by inhibiting the adhesion of C. albicans cells. This response may be associated with the down-regulation of adhesion-related genes ALS1, ALS3 and BCR1. The inhibitory serum component is protease-resistant and heat stable.
白色念珠菌可在静脉导管上形成生物膜;这一过程在导管感染的发病机制中起关键作用。本研究评估了人血清(HS)对白色念珠菌生物膜形成及体外黏附相关基因表达的影响。一株白色念珠菌实验室菌株(ATCC90028)和三株临床菌株分别在添加HS的RPMI 1640培养基或仅含RPMI 1640的培养基(作为对照)中培养24小时。通过活细胞电影分析仪和XTT还原试验监测四株菌株生物膜细胞的生长情况。在三个时间点(60分钟、90分钟和24小时)通过RT-PCR分析黏附相关基因BCR1、ALS1、ALS3、HWP1和ECE1的表达。
在黏附阶段,白色念珠菌细胞在含HS的RPMI培养基中保持布朗运动,直至形成大量芽管。在对照组中,白色念珠菌细胞迅速黏附于反应板底部。与RPMI 1640相比,添加3% - 50% HS的培养基导致生物膜形成显著减少(所有p < 0.001)。然而,HS的存在对预先黏附的生物膜无显著抑制作用(所有p > 0.05)。热灭活和蛋白酶K预处理的HS也抑制生物膜形成。50% HS的存在对白色念珠菌的浮游生长无显著影响(p > 0.05)。在三个时间点,HS抑制ALS1和ALS3基因的表达,并促进HWP1和ECE1基因的表达。仅在90分钟时观察到BCR1显著上调。
人血清通过抑制白色念珠菌细胞的黏附来减少生物膜形成。这种反应可能与黏附相关基因ALS1、ALS3和BCR1的下调有关。血清中的抑制成分具有蛋白酶抗性和热稳定性。