三七对耐甲氧西林金黄色葡萄球菌细胞壁的作用及机制  

Effect and mechanism of Panax Notoginseng on methicillin-resistant staphylococcus aureus cell wall

作　　者：冯小玉 杨伟峰[2] 夏玉文 温博 张璐璐 安洪萨 葛锋[3] 谭勇[1] FENG Xiaoyu;YANG Weifeng;XIA Yuwen;WEN Bo;ZHANG Lulu;AN Hongsa;GE Feng;TAN Yong(Institute of Basic Research in Clinical Medicine,China Academy of Chinese Medical Sciences,Beijing100700,China;Medical Experimental Center,China Academy of Chinese Medical Sciences,Beijing100700,China;School of Life Science and Technology,Kunming University of Technology Kunming 650500)

机构地区：[1]中国中医科学院中医临床基础医学研究所,北京100700 [2]中国中医科学院医学实验中心,北京100700 [3]昆明理工大学生命科学与技术学院,昆明650500

出　　处：《北京中医药》2024年第6期655-660,共6页Beijing Journal of Traditional Chinese Medicine

基　　金：中国中医科学院自主选题项目(Z0735);中国中医科学院科技创新工程中医临床基础学科创新团队项目(CI2021B003);中国中医科学院科技创新工程中医临床基础学科重大攻关项目子课题(CI2021A00704-2);云南省科学技术厅课题(Z0797)。

摘　　要：目的 探究三七(PN)对耐甲氧西林金黄色葡萄球菌(MRSA)细胞壁的作用及机制。方法 实验分对照组(Control组)和PN组,Control组MRSA未干预,PN组MRSA被2 mg/mLPN干预24 h。酶联免疫吸附试验(ELISA)检测PN对MRSA细胞壁肽聚糖(PGN)和脂磷壁酸(LTA)的影响;透射电镜(TEM)观察MRSA细胞壁厚度;转录组测序技术(RNA-Seq)筛选PN干预MRSA的药效相关基因,通过基因本体(GO)与京都基因与基因组百科全书(KEGG)对其进行富集分析;实时荧光定量PCR技术验证PN干预MRSA的细胞壁相关基因。结果 与Control组比较,PN组PGN、LTA降低(P<0.05);PN组处于分裂期的细菌数量增多,体积变大,细菌分裂隔膜模糊,细胞壁厚度薄(P<0.05)。2组间有552个差异基因,相较于Control组,PN组285个基因上调、267个基因下调;差异基因涉及的生物过程(BP)主要富集在氧化还原、前体代谢产物和能量产生、细胞氨基酸分解代谢、有氧呼吸过程,细胞组分(CC)主要富集在细胞解剖实体、细胞器官、非膜界细胞器,分子功能(MF)主要富集在氧化还原酶活性、结构分子活性、电子转移活性;差异基因共富集85条信号通路,其中包括脂肪酸、赖氨酸降解;差异基因MraY、MurD、MurC、FemA、FemB、sgtB与MRSA细胞壁相关。与Control组比较,PN组MraY、MurD、FemA、FemB表达降低(P<0.05),MurC、sgtB表达升高(P<0.05)。结论 PN可抑制MRSA细胞壁,与降低细胞壁相关基因表达和促进脂肪酸、赖氨酸降解等有关。Objective To explore the effect and mechanism of panax notoginseng(PN)on methicillin-resistant staphylococcus aureus(MRSA)cell wall.Methods There were PN treatment group and control group in this study.MRSA in control group was not intervened,while MRSA in PN group was intervened by 2 mg/mLPN for 24 hours.The effects of PN on wall peptidoglycan(PGN)and wall phosphate(LTA)in MRSA cells were detected using ELISA.The effect of PN on the thickness of MRSA cell wall was observed through transmission electron microscopy.Genes related to the efficacy of PN were screened using RNA Seq technology,then these genes were verified by RT-PCR.Results Compared with the control group,the levels of PGN and LTA in MRSA in PN group were significantly reduced(P<0.05).In PN group,the number and volume of bacteria in dividing stage increased,the dividing membrane of bacteria was blurred and the cell wall thickness was thin(P<0.05).There were 552 different genes between the two groups.Compared with the control group,285 genes in PN group were up-regulated and 267 genes were down-regulated.Biological processes(BP)related to differential genes are mainly concentrated in redox,precursor metabolites and energy production,cellular amino acid catabolism and aerobic respiration,cellular components(CC)are mainly concentrated in cellular anatomical entities,cellular organs and non-membrane organelles,and molecular functions(MF)are mainly concentrated in redox enzyme activity,structural molecular activity and electron transfer activity.The differential genes enriched 85 signal pathways,including fatty acid and lysine degradation.The differential genes MraY,MurD,MurC,FemA,FemB and sgtB are related to MRSA cell wall.Compared with the control group,the expressions of MraY,MurD,FemA and FemB in PN group decreased(P<0.05),while the expressions of MurC and sgtB increased(P<0.05).Conclusion PN exerts the therapeutic effect on MRSA by inhibiting its cell wall which is related to reducing the expression of cell wall-related genes and promoting the degrada

关 键 词：三七 耐甲氧西林金黄色葡萄球菌 细胞壁 抑制作用 肽聚糖 脂磷壁酸 

分 类 号：R285[医药卫生—中药学]