摘要
以携带多重耐药基因cfr的IncX4型质粒pSD11为试验材料,研究小檗碱、黄芩苷、芦荟大黄素、姜黄素4种中药单体对耐药质粒稳定性的影响,为中药单体消除病原菌耐药性提供理论依据.通过测定最小抑菌浓度(MIC)及药物作用下携带质粒pSD11的大肠杆菌的生长曲线和生长动力学参数,分析中药单体的抑菌效果.结合实时荧光定量PCR(RTPCR)及平板计数研究药物对质粒稳定性的影响,同时检测胞外乳酸脱氢酶和碱性磷酸酶的含量分析中药单体对细胞壁、细胞膜的影响.结果显示,小檗碱、黄芩苷、芦荟大黄素对大肠杆菌MG1655/pSD11(Escherichia coli MG1655/pSD11)的MIC均为1280μg/mL,姜黄素>160μg/mL.4种中药单体对E.coli MG1655/pSD11具有一定的抑菌效果,生长动力学参数结果表明1/2 MIC小檗碱和黄芩苷显著延长E.coli MG1655/pSD11的滞后期;1/2 MIC姜黄素和1/2、1/4MIC芦荟大黄素极显著降低E.coli MG1655/pSD11的最大生长速率、延长时代时间.经1/2和1/4 MIC的小檗碱和黄芩苷处理后,质粒pSD11的稳定性极显著降低,且细菌胞外乳酸脱氢酶和碱性磷酸酶含量增加.本研究表明,4种中药单体均能有效抑菌,且小檗碱和黄芩苷能够降低IncX4型质粒pSD11的稳定性.(图5表3参25附图1)
The IncX4 plasmid pSD11,which carries the multidrug resistance gene cfr was used to study the effects of berberine,baicalin,aloe-emodin,and curcumin on stability of drug-resistant plasmids,thus providing a theoretical basis for the elimination of drug resistance in pathogenic bacteria by traditional Chinese medicine(TCM)monomers.The antibacterial effect of the four TCM monomers was analyzed by measuring the minimum inhibitory concentration(MIC),growth curve,and growth kinetic parameters of Escherichia coli strain MG1655/pSD11(E.coli MG1655/pSD11)while exposed to drugs.Real-time Quantitative PCR(RT-PCR)and plate counting were used to investigate the effects of the four TCM monomers on plasmid stability.Additionally,the extracellular levels of lactate dehydrogenase and alkaline phosphatase were determined in order to investigate the monomer’s effects on the cell wall and cell membrane.Results suggest that the four TCM monomers have an antibacterial effect on E.coli MG1655/pSD11 with an MIC value of 1280μg/mL for berberine,baicalin,and aloeemodin and>160μg/mL for curcumin.Meanwhile,investigation of growth kinetic parameters of E.coli MG1655/pSD11 indicated that berberine and baicalin(1/2 MIC)significantly prolonged the lag time,whereas curcumin(1/2 MIC)and aloe-emodin(1/2,1/4 MIC)significantly reduced the maximum growth rate and prolonged the generation time.After treatment with 1/2 and 1/4 MIC of berberine and baicalin,the stability of plasmid pSD11in E.coli MG1655/pSD11 was significantly reduced,while the extracellular levels of lactate dehydrogenase and alkaline phosphatase increased.In summary,our findings indicate that the TCM monomers effectively inhibit bacteria,while berberine and baicalin were shown to reduce stability of the IncX4 plasmid pSD11.
作者
陈璐
邓翔文
郑婷婷
黄小红
邓辉
CHEN Lu;DENG Xiangwen;ZHENG Tingting;HUANG Xiaohong;DENG Hui(University Key Laboratory for Integrated Chinese Traditional and Western Veterinary Medicine and Animal Healthcare in Fujian Province,Fuzhou 350002,China;Fujian Key Laboratory of Traditional Chinese Veterinary Medicine and Animal Health,Fujian Agriculture and Forestry University,Fuzhou 350002,China)
出处
《应用与环境生物学报》
CAS
CSCD
北大核心
2023年第1期57-63,共7页
Chinese Journal of Applied and Environmental Biology
基金
国家自然科学基金青年科学基金项目(31602106)
福建省重大专项专题项目(2021NZ029008)
福建农林大学科技创新专项基金项目(CXZX2020062A,CXZX2020063A)资助
