中华生物医学工程杂志
中華生物醫學工程雜誌
중화생물의학공정잡지
CHINESE JOURNAL OF BIOMEDICAL ENGINEERING
2012年
4期
265-268
,共4页
念珠菌,白色%生物膜%CPH1%EFG1%基因表达
唸珠菌,白色%生物膜%CPH1%EFG1%基因錶達
념주균,백색%생물막%CPH1%EFG1%기인표체
Candida albicans%biofilms%CPH1%EFG1%Gene expression
目的 检测游离态及不同时期生物膜态白念珠菌转录因子CPH1和EFG1的表达,探讨其在生物膜形成过程中的作用.方法 应用激光共聚焦显微镜观察白念珠菌质控株ATCC90028和临床分离株14215于聚乙烯片上黏附生长24 h后的生物膜形态.分别提取6株白念珠菌临床分离株13860、13874、14127、14371、14215、14533和质控株ATCC90028游离态以及早期(聚乙烯片上黏附生长4h)、中期(聚乙烯片上黏附生长12h)、晚期(聚乙烯片上黏附生长24 h)生物膜态的总RNA,应用荧光定量PCR测定其CPH1和EFG1基因的表达.结果 聚乙烯片上黏附生长24 h后,激光共聚焦显微镜下可见质控株ATCC90028多为单层的孢子细胞黏附;白念珠菌临床分离株14215形成菌丝态为主,具有三维结构的生物膜.相对于游离态,白念珠菌质控株ATCC90028在生物膜形成的早中晚期转录因子CPH1和EFG1表达均下调.临床分离株转录因子CPH1在游离态、生物膜形成的早中晚期时表达差异均无统计学意义(均P＜0.05).而与游离态相比,临床分离株转录因子EFG1基因的相对表达量在生物膜形成早期及中期均上调[0.141(0.029 ～ 0.212)、0.252(0.103 ～ 0.943)比0.077(0.018 ～ 0.113),均P＜0.05],在生物膜形成晚期则无明显改变(P＞0.05),相对表达量为0.091(0.024 ～ 0.354).结论 转录因子EFG1在白念珠菌临床分离株生物膜形成过程中有重要的调控作用.
目的 檢測遊離態及不同時期生物膜態白唸珠菌轉錄因子CPH1和EFG1的錶達,探討其在生物膜形成過程中的作用.方法 應用激光共聚焦顯微鏡觀察白唸珠菌質控株ATCC90028和臨床分離株14215于聚乙烯片上黏附生長24 h後的生物膜形態.分彆提取6株白唸珠菌臨床分離株13860、13874、14127、14371、14215、14533和質控株ATCC90028遊離態以及早期(聚乙烯片上黏附生長4h)、中期(聚乙烯片上黏附生長12h)、晚期(聚乙烯片上黏附生長24 h)生物膜態的總RNA,應用熒光定量PCR測定其CPH1和EFG1基因的錶達.結果 聚乙烯片上黏附生長24 h後,激光共聚焦顯微鏡下可見質控株ATCC90028多為單層的孢子細胞黏附;白唸珠菌臨床分離株14215形成菌絲態為主,具有三維結構的生物膜.相對于遊離態,白唸珠菌質控株ATCC90028在生物膜形成的早中晚期轉錄因子CPH1和EFG1錶達均下調.臨床分離株轉錄因子CPH1在遊離態、生物膜形成的早中晚期時錶達差異均無統計學意義(均P＜0.05).而與遊離態相比,臨床分離株轉錄因子EFG1基因的相對錶達量在生物膜形成早期及中期均上調[0.141(0.029 ～ 0.212)、0.252(0.103 ～ 0.943)比0.077(0.018 ～ 0.113),均P＜0.05],在生物膜形成晚期則無明顯改變(P＞0.05),相對錶達量為0.091(0.024 ～ 0.354).結論 轉錄因子EFG1在白唸珠菌臨床分離株生物膜形成過程中有重要的調控作用.
목적 검측유리태급불동시기생물막태백념주균전록인자CPH1화EFG1적표체,탐토기재생물막형성과정중적작용.방법 응용격광공취초현미경관찰백념주균질공주ATCC90028화림상분리주14215우취을희편상점부생장24 h후적생물막형태.분별제취6주백념주균림상분리주13860、13874、14127、14371、14215、14533화질공주ATCC90028유리태이급조기(취을희편상점부생장4h)、중기(취을희편상점부생장12h)、만기(취을희편상점부생장24 h)생물막태적총RNA,응용형광정량PCR측정기CPH1화EFG1기인적표체.결과 취을희편상점부생장24 h후,격광공취초현미경하가견질공주ATCC90028다위단층적포자세포점부;백념주균림상분리주14215형성균사태위주,구유삼유결구적생물막.상대우유리태,백념주균질공주ATCC90028재생물막형성적조중만기전록인자CPH1화EFG1표체균하조.림상분리주전록인자CPH1재유리태、생물막형성적조중만기시표체차이균무통계학의의(균P＜0.05).이여유리태상비,림상분리주전록인자EFG1기인적상대표체량재생물막형성조기급중기균상조[0.141(0.029 ～ 0.212)、0.252(0.103 ～ 0.943)비0.077(0.018 ～ 0.113),균P＜0.05],재생물막형성만기칙무명현개변(P＞0.05),상대표체량위0.091(0.024 ～ 0.354).결론 전록인자EFG1재백념주균림상분리주생물막형성과정중유중요적조공작용.
Objective To detect the expression of CPH1 and EFG1,the transcription factors,in planktonic stage and various phases during biofilm stage of Candida albicans,and to explore their roles in biofilm formation.Methods Confocal scanning laser microscope was employed for morphological examination of biofilm formation of ATCC90028,the strain for quality control,and the clinically isolated strain 14215 adhering to polyethylene film for 24 hours.The total RNA of 6 clinically isolated strains (13860,13874,14127,14371,14215 and 14533) and ATCC90028 during planktonic stage,and at early (4-hour adhesion to polyethylene film),intermediate ( 12-hour adhesion to polyethylene film) and late phase (24-hour adhesion to polyethylene film) during biofilm stage was extracted respectively.Expression of CPH1 and EFG1 genes was determined using fluorescent quantitative polymerase chain reaction.Results After the 24-hour adhesion to polyethylene films,the ATCC90028 strain appeared mostly as blastospores in a single layer while the clinically isolated strain 14215 was in the form of hypha producing a three-dimensional biofilm.Expression of CPH1 and EFG1 was down-regulated in the early,intermediate and late phases during biofilm formation,compared with planktonic stage of the ATCC90028 strain.CPH1 expression in the strain 14215 did not differ statistically between the planktonic stage and early,intermediate and late phases during biofilm formation (all P＞0.05).Contrarily,EFG1 expression was up-regulated at early and intermediate stages during biofilm formation as compared with that during planktonic stage [ (0.141 (0.029-0.212) and 0.252 (0.103-0.943) vs 0.077 (0.018-0.113),all P＜0.05].No significant change in EFG1 expression was detected at the late phase during biofilm formation [0.091 (0.024- 0.354),P＞0.05].Conclusion Transcription factor EFG1 may play a critical role in the regulation of biofilm formation in clinically isolated Candida albicans strains.
