(R)-(+)-长叶薄荷酮对耐药结核分枝杆菌抑制作用及其生物被膜调控基因的影响  

Inhibitory effects of(R)-(+)-pulegone on drug-resistant Mycobacterium tuberculosis and itsbiofilm-regulating genes

作　　者：王晓娟 熊文娟 宫海燕 WANG Xiaojuan;XIONG Wenjuan;GONG Haiyan(Department of Laboratory,The Fifth Affiliated Hospital of Xinjiang Medical University,Urumqi 830011,China;Centerof Medical Laboratory,the First Dongguan Affiliated Hospital of Guangdong Medical University,Dongguan Guangdong 523000,China)

机构地区：[1]新疆医科大学第五附属医院检验科,乌鲁木齐830011 [2]广东医科大学附属东莞第一医院医学检验中心,广东东莞523000

出　　处：《新疆医科大学学报》2024年第3期409-413,420,共6页Journal of Xinjiang Medical University

基　　金：2020年省部共建中亚高发病成因与防治国家重点实验室开放课题项目(SKL-HIDCA-2020-WF3)。

摘　　要：目的 研究(R)-(+)-长叶薄荷酮对耐药结核分枝杆菌体外抑制作用及其生物被膜调控基因Rv0024、Rv2872、Ra1362的影响。方法 通过聚合酶链反应(Polymerase chain reaction, PCR)筛选含生物被膜调控基因Rv0024、Rv2872、Ra1362的耐药结核分枝杆菌,采用刃天青显色法和液体培养计数法测定(R)-(+)-长叶薄荷酮对生物被膜介导的耐药结核分枝杆菌的最小抑菌浓度(Minimal inhibit concentration, MIC)和最低杀菌浓度(Minimum bactericidal concentration, MBC),应用反转录-聚合酶链式反应(Reverse transcription-polymerase chain reaction, RT-PCR)检测(R)-(+)-长叶薄荷酮对耐药结核分枝杆菌生物被膜调控基因Rv0024、Rv2872、Ra1362的影响。结果 在3×10^(7) CFU/mL和3×10^(6) CFU/mL菌液浓度下,刃天青显色法测得(R)-(+)-长叶薄荷酮对耐药结核分枝杆菌的MIC和MBC分别为14.79、29.59 mg/mL;液体培养计数法测得(R)-(+)-长叶薄荷酮对耐药结核分枝杆菌的MIC_(90)和MBC分别为19.89、24.62 mg/mL。RT-PCR法结果表明长叶薄荷酮可抑制Rv0024、Ra1362的表达,促进Rv2872的表达(P<0.05)。结论 (R)-(+)-长叶薄荷酮通过调控耐药结核分枝杆菌生物被膜主要基因Rv0024、Rv2872、Ra1362的表达,达到抑制耐药结核分枝杆菌的生长的作用。Objective This study investigated in vitro inhibitory effects of(R)-(+)-pulegone on drug-resistant Mycobacterium tuberculosis and its influence on biofilm-regulating genes Rv0024,Rv2872and Ra1362.Methods Drug-resistant Mycobacterium tuberculosis strains containing the biofilm-regulating genes Rv0024,Rv2872 and Ra1362 were screened using polymerase chain reaction(PCR).The minimum inhibitory concentration(MIC)and minimum bactericidal concentration(MBC)of(R)-(+)-pulegone against biofilm-mediated drug-resistant Mycobacterium tuberculosis were determined using the methylene blue dye method and liquid culture counting method,respectively.Reverse transcription-polymerase chain reaction(RT-PCR)was employed to assess the impact of(R)-(+)-pulegone on the expression of biofilm-regulating genes Rv0024,Rv2872 and Ra1362.Results The MIC and MBC of(R)-(+)-pulegone against drug-resistant Mycobacterium tuberculosis were found to be 14.79 mg/mL and 29.59 mg/mL,respective-ly,using the methylene blue dye method at a bacterial density of 3×10^(7) CFU/mL and 3×10^(6) CFU/mL.The MIC_(90) and MBC of(R)-(+)-pulegone determined by the liquid culture counting method were 19.89 mg/mL and 24.62 mg/mL,respectively.The results of RT-PCR showed that long leaved menthone could inhibit the expression of Rv0024 and Ra1362,and promote the expression of Rv2872(P<0.05).Conclusions(R)-(+)-pulegone inhibits the growth of drug-resistant Mycobacterium tuberculosis by regu-lating the expression of the main genes Rv0024,Rv2872and Ra1362 in the biofilm of drug-resistant Myco-bacterium tuberculosis.

关 键 词：(R)-(+)-长叶薄荷酮 耐药结核分枝杆菌 抑制作用 生物被膜 调控基因 

分 类 号：R378[医药卫生—病原生物学]