雏菊叶龙胆酮的抑菌作用研究  

Study on the antibacterial activity of Bellidifolin

作　　者：陈雪晴 张峥嵘[2] 马丽 孙加燕 查元 慈傲特 朱金燕[1] CHEN Xueqing;ZHANG Zhengrong;MA Li;SUN Jiayan;ZHA Yuan;CI Aote;ZHU Jinyan(School of Pharmacy,Anqing Medical College,Anqing 246052,Anhui,China;School of Chinese Medicine,Anhui University of Chinese Medicine,Hefei 230012,Anhui,China)

机构地区：[1]安庆医药高等专科学校药学院,安徽安庆246052 [2]安徽中医药大学中医学院,安徽合肥230012

出　　处：《右江民族医学院学报》2025年第1期44-49,共6页Journal of Youjiang Medical University for Nationalities

基　　金：安徽省高校自然科学研究项目(2022AH052544,2023AH053424)。

摘　　要：目的探讨雏菊叶龙胆酮的抑菌作用及机制。方法雏菊叶龙胆酮作用大肠杆菌、金黄色葡萄球菌和白色念珠菌后,通过微量稀释法测定最低抑菌浓度(MIC)、最低杀菌浓度(MBC),进行耐药性诱导,以荧光染料4’,6-二脒基-2-苯基吲哚(DAPI)、碘化丙啶(PI)检测菌的死活情况,扫描电镜观察细胞形态,利用RT-PCR分析大肠杆菌外输泵基因AcrA、AcrB的表达。结果雏菊叶龙胆酮对大肠杆菌、金黄色葡萄球菌和白色念珠菌MIC分别为50μg/mL、100μg/mL、100μg/mL;MBC分别为200μg/mL、250μg/mL、250μg/mL。作用于金黄色葡萄球菌和白色念珠菌时随着诱导代数的增加,未出现耐药性。Live/Dead染色结果显示雏菊叶龙胆酮能够透过细胞膜,且随着药物浓度的增加,死亡的菌细胞数逐渐增加。扫描电镜显示雏菊叶龙胆酮对3种菌的形态具有不同程度的影响,其中大肠杆菌细胞菌体的表面形态出现明显的溶解和坍塌,随着浓度的增加,大部分菌体严重皱缩、干瘪、扭曲变形、菌体表面凹凸不平,且可降低大肠杆菌其外排泵基因AcrA、AcrB的相对表达量(P<0.01)。结论雏菊叶龙胆酮对大肠杆菌、金黄色葡萄球菌和白色念珠菌均具有一定抑菌作用,细菌对其不易产生耐药性,其中对大肠杆菌的抑菌活性最佳,主要通过损伤大肠杆菌细胞形态,改变细菌细胞膜的通透性。且对大肠杆菌耐药菌的外排泵基因表达量具有抑制作用。Objective To investigate the antibacterial activity and mechanism of Bellidifolin.Methods After treating Escherichia coli(E.coli),Staphylococcus aureus(S.aureus),and Candida albicans(C.albicans)with Bellidifolin,the minimum inhibitory concentration(MIC)and minimum bactericidal concentration(MBC)were determined by the microdilution method.Drug resistance induction was conducted,and the viability of bacteria was detected using fluorescent dyes 4’,6-diamidino-2-phenylindole(DAPI)and propidium iodide(PI).The cell morphology was observed by scanning electron microscopy(SEM),and the expression of the efflux pump genes AcrA and AcrB in E.coli was analyzed by RT-PCR.Results The MIC of Bellidifolin against E.coli,S.aureus and C.albicans was 50μg/mL,100μg/mL and 100μg/mL,respectively.The MBCs were 200μg/mL,250μg/mL and 250μg/mL,respectively.No drug resistance was observed in S.aureus and C.albicans with increasing induction generations.The results of live/dead staining showed that Bellidifolin could penetrate the cell membrane,and the number of dead bacterial cells increased with the concentration of the drug.SEM showed that it had different degrees of influence on the morphology of the three kinds of bacteria,among which the surface morphology of E.coli cells showed obvious dissolution and collapse.With the increase of concentration,most of the bacteria were seriously shrunken,shriveled,distorted and uneven on the surface.Furthermore,Bellidifolin reduced the relative expression of the efflux pump genes AcrA and AcrB in E.coli(P<0.01)Conclusion Bellidifolin has a certain antibacterial effect against E.coli,S.aureus,and C.albicans.Bacteria are not prone to develop resistance to it.Among them,it has the best antibacterial activity against E.coli,mainly by damaging the cell morphology of E.coli and changing the permeability of the bacterial cell membrane,and has a certain inhibitory effect on the expression of the efflux pump genes of drug-resistant E.coli.

关 键 词：雏菊叶龙胆酮 抑菌作用 大肠杆菌 金黄色葡萄球菌 白色念珠菌 

分 类 号：R285[医药卫生—中药学]