S-腺苷甲硫氨酸合成酶参与肺炎支原体生物被膜形成  

S-adenosylmethionine synthase is involved in Mycoplasma pneumoniae biofilm formation

作　　者：李婷婷 陈春艳 余斓 丁伟艳 丁楠 李水红 朱翠明 LI Ting-ting;CHEN Chun-yan;YU Lan;DING Wei-yan;DING Nan;LI Shui-hong;ZHU Cui-ming(Hunan Provincial Key Laboratory for Special Pathogens Prevention and Control,Institute of Pathogenic Biology,Medical College,University of South China,Hengyang 421001,China;Clinical Laboratory of the Central Hospital of Xiangtan,Xiangtan 411100,China;Department of Blood Transfusion,the First Afiliated Hospital of University of South China,Hengyang 421001,China)

机构地区：[1]南华大学衡阳医学院病原生物学研究室,特殊病原体防控湖南省重点实验室,衡阳421001 [2]湘潭市中心医院检验科,湘潭411100 [3]南华大学附属第一医院输血科,衡阳421001

出　　处：《中国人兽共患病学报》2021年第5期404-409,共6页Chinese Journal of Zoonoses

基　　金：国家自然科学基金(No.31970177);湖南省自然科学基金(No.2019JJ40253);湖南省教育厅重点项目(No.18A234);湖南省研究生科研创新项目(No.CX2018B622)联合资助。

摘　　要：目的了解S-腺苷甲硫氨酸(SAM)合成酶在肺炎支原体(Mp)生物被膜(BF)形成中的作用,初步探讨MpBF的形成机制。方法在24孔细胞培养板中培养Mp,结晶紫染色后半定量分析Mp M129标准菌株和22株临床分离株的BF形成能力。荧光定量逆转录PCR(qRT-PCR)分析编码SAM合成酶的基因(metX)在BF形成能力强和形成能力弱的Mp菌株中的转录水平。分别用西奈芬净和迷迭香酸处理Mp,qRT-PCR检测metX的mRNA相对表达水平,结晶紫染色半定量分析MpBF的形成量。结果13株Mp菌株(56.5%,13/23)可在细胞培养板中显著形成BF。3株形成MpBF菌株的metX mRNA转录水平均高于3株不能形成BF的Mp菌株(P<0.05)。西奈芬净可抑制MpBF的产生和metX mRNA的转录水平;迷迭香酸可促进MpBF形成及metX mRNA的表达。结论metX mRNA的转录水平与MpBF的形成相关,SAM合成酶可能参与MpBF的形成。This study aimed to explore the influence of S-adenosylmethionine synthase on Mycoplasma pneumoniae biofilm production,to better understand the underlying mechanisms.M.pneumoniae Mp129 strains and 22 clinical isolates were cultured in 24-well plates.After crystal violet staining,M.pneumoniae biofilms were observed under a light microscope and semi-quantitatively analyzed according to the OD value at 570 nm.Quantitative reverse transcription polymerase chain reaction(RT-qPCR)was used to detect the mRNA expression levels of the S-adenosylmethionine synthase gene(metX)in three M.pneumoniae strains with biofilm formation and three strains with weak or no biofilm formation.The effects of sinefungin and rosmarinic acid on the metX mRNA transcription level and biofilm formation in the M.pneumoniae strains was evaluated in vitro.Among the 23 M.pneumoniae strains,13 strains formed biofilms,whereas 10 isolates did not form biofilms or produced only a small amount of biofilms in 24-well plates.The mRNA expression level of metX was much higher in the M.pneumoniae strains with biofilm formation than in those with weak or no biofilm formation(P<0.05).Furthermore,the metX transcription level and mycoplasmal biofilm formation were significantly inhibited by sinefungin and increased by rosmarinic acid.The formation of M.pneumoniae biofilm correlated with the metX mRNA transcription level;therefore,S-adenosylmethionine synthase may be involved in the formation of M.pneumoniae biofilm.

关 键 词：肺炎支原体 生物被膜 S-腺苷甲硫氨酸合成酶 西奈芬净 迷迭香酸 

分 类 号：R375.2[医药卫生—病原生物学]