可耐受高温对黑曲霉生物膜体外模型构建的影响  

In Vitro Efficacy of Continuous Mild High Temperature on the Biofilm Formation of Aspergillus Niger

作　　者：曾荣[1] 童建波 刘宇甄 陈青[2] 林彤[1] 李岷 吕桂霞[1] 

机构地区：[1]中国医学科学院北京协和医学院皮肤病研究所,南京210042 [2]江苏省血液中心,南京210042 [3]江苏省皮肤性病学分子生物学重点实验室,南京210042

出　　处：《中国医学科学院学报》2017年第6期762-767,共6页Acta Academiae Medicinae Sinicae

基　　金：国家自然科学基金(81502739;81371750);江苏省自然科学基金(BK20150068);北京协和医学院青年基金(3332016108);中国医学科学院医学与健康科技创新工程项目(CIFMS-2017-I2M-1-017);973项目(2013CB531605)~~

摘　　要：目的在可耐受高温作用下构建黑曲霉生物膜模型,探讨可耐受高温效应对其形成结构及活力的影响。方法对比37℃下,分别采用共聚焦激光扫描显微镜和甲基四氮盐法探索在不同高温效应下(39℃或41℃)体外黑曲霉生物膜形成过程中的多个时间观察点(4、8、10、16、24、48和72 h)的超微结构差异及活力变化。结果与37℃相比,在4 h时,温度较高的39℃组和41℃组黏附率均显著降低(t=8.603,P=0.047;t=14.550,P=0.008),黏附浓度也均显著降低(t=8.315,P=0.0021;t=11.748,P=0.0015);在8 h时,温度越高,出芽率越低(t=8.906,P=0.049;t=23.310,P=0.009)、菌丝长度越短(t=35.118,P=0.039;t=63.450,P=0.006);在10~16 h菌丝生长阶段,温度越高菌丝极性生长破坏越严重,菌丝越弯曲,且分枝越多,活力越高(t=24.262,P=0.038;t=7.556,P=0.031);24 h时,温度越高生物膜厚度越薄(t=2.375,P=0.043,t=5.263,P=0.031),菌丝方向性越差,活力越低(t=24.262,P=0.038;t=7.556,P=0.031)。结论可耐受高温作用能够在一定程度上抑制黑曲霉生物膜的形成及其活力。Objective To investigate whether continuous mild high temperature（increased temperature without causing significant damage to host cells） can inhibit the biofilm formation of Aspergillus niger（A.niger）and its vitality.Methods A.niger biofilms were formed on a coverslip in 24-well tissue culture plate and werechecked at the time points 4,8,10,16,24,48 and 72 hours.Confocal laser scanning microscopy（CLSM）was used to image and quantify A.niger biofilm formation under three different continuous mild high temperatures at 37℃,39℃,and 41℃.Furthermore,2,3-bis（2-methoxy-4-nitro-5-sulfophenyl）-2 H-tetrazolium-5-carboxanilide（XTT） assay was used to quantify the dynamic growth of A.niger biofilm under the above conditions.Results Compared with the culture condition 37℃,CLSM analysis at 39℃ or 41℃ showed that higher temperature induced later germination at 4 hours（t=8.603,P=0.047;t=14.550,P=0.008）,poorer hyphal elongation at 8 hours（t=35.118,P=0.039;t=63.450,P=0.006）,poorer polar growth,and reduced biofilm thickness from 10 to 24 hours.The XTT assay showed that higher temperature（39℃ or 41℃） lead to lower vitality at 10 hours,higher vitality at 16 hours,but finally lower vitality from 24 to 72 hours（t=24.262,P=0.038;t=7.556,P=0.031）.Conclusion Continuous mild high temperature may have a negative regulatory effect on biofilm formation of A.niger and its vitality.

关 键 词：黑曲霉 生物膜 高温作用 

分 类 号：R379.6[医药卫生—病原生物学]