小檗碱对耐甲氧西林金黄色葡萄球菌全基因组转录水平的影响研究  

Effect of berberine on the whole genome transcription of methicillin-resistant Staphylococcus aureus

作　　者：周芳芳 王蕾[1] 王微[1] 卢培[1] Zhou Fangfang;Wang Lei;Wang Wei;Lu Pei(Department of Clinical Laboratory,Shanghai Eighth People′s Hospital,Shanghai 200235,China)

机构地区：[1]上海市第八人民医院检验科,上海200235

出　　处：《中华检验医学杂志》2020年第5期576-581,共6页Chinese Journal of Laboratory Medicine

摘　　要：目的通过转录组测序(RNA-seq)分析小檗碱对耐甲氧西林金黄色葡萄球菌USA300基因组表达谱的影响,研究小檗碱对耐甲氧西林金黄色葡萄球菌(MRSA)的抑菌机制。方法基础研究。本研究使用标准菌株USA300,该菌株为中科院上海药物所蓝乐夫课题组2019年所赠送。用1/2最低抑菌浓度(MIC)(64 mg/L)和1/8MIC(16 mg/L)浓度的小檗碱处理和无小檗碱处理耐甲氧西林金黄色葡萄球菌USA300,分析USA300差异基因表达情况。将差异基因筛选条件设为:P值<0.05,且差异倍数>2。采用SPSS 20.0进行统计分析。结果通过测序结果发现,高浓度组与正常对照组相比,共有754个差异基因表达,其中上调基因主要为lukD、lukE、ssaA等,下调基因主要为lukF-PV、lukS-PV、clfA、splB^splF、sspA等;低浓度组与正常对照组相比,仅有19个差异基因表达,其中上调基因为speG、scpB、rpsQ等,下调基因为betA、bccT等。结论本研究USA300转录组基因表达情况,了解经小檗碱作用后差异基因表达情况,推测小檗碱主要通过调控细菌细胞壁水解功能、白细胞毒素及其他毒力因子的表达发挥对MRSA的抑菌作用。Objective The work aims to analyze the effect of berberine on the transcriptome-feature of the methicillin-resistant Staphylococcus aureus(MRSA)USA300 by using transcriptome sequencing(RNA-seq methods)methods and to investigate the bacteriostatic mechanism of berberine on MRSA.Methods Basic research.The standard strain USA300 was used in this study,which was a honorable gift from Dr.Lan Lefu in Shanghai Institute of medicine,Chinese Academy of Sciences in 2019.The differential gene expression profiles of berberine treated[1/2 MIC(64 mg/L)and 1/8 MIC(16 mg/L),respectively]and untreated USA300 were analyzed.The screening conditions for differentially expressed genes were set as:P-Value<0.05,and the|Fold-change|>2.SPSS 20.0 was used to analyze the result.Results Sequencing results showed that there were 754 differentially expressed genes in the high concentration group compared with the normal control group,of which the up-regulated genes were lukD,lukE,ssaA,etc.,while the down-regulated genes were lukF-PV,lukS-PV,clfA,splB-splF,sspA,etc..Only 19 genes were differentially expressed in the low concentration group compared with the normal control group,of which the up-regulated genes were speG,scpB,rpsQ,etc.,while the down-regulated genes were beta,bccT,etc.Conclusions In this study,the gene expression of USA300 transcriptome was studied to understand the differential gene expression after berberine treatment.It was speculated that berberine played a role in inhibiting MRSA by regulating the hydrolysis of bacterial cell wall,the expression of leucotoxin and other virulence factors.

关 键 词：小檗碱 耐甲氧西林金黄色葡萄球菌 序列分析 RNA 基因表达谱 

分 类 号：R285.5[医药卫生—中药学]