3种中药纳米乳剂对耳念珠菌深部感染小鼠的抗菌效果
Antibacterial effects of three types of traditional Chinese medicine nanoemulsions on mice with deep-seated Candida auris infection
陈春屹 1余曦晨 1王迪 1廖万清 2罗罡 1康颖倩1
作者信息
- 1. 贵州医科大学基础医学院微生物学教研室,贵州贵阳 550025;贵州省普通高校病原生物学特色重点实验室,贵州贵阳 550025
- 2. 上海市医学真菌分子生物学重点实验室,上海 200003;海军军医大学长征医院皮肤科,上海 200003
- 折叠
摘要
目的 比较3种中药纳米乳剂对耳念珠菌感染小鼠深部感染的效果.方法 取茶树精油(TTO)、艾纳香油(BBO)、余甘子、白芨、白茯苓、白芷及柠檬酸,采用高压均质法及超声波法制备中药纳米乳剂(简称纳米剂)1(TTO、BBO及水相全部药物组成)、纳米剂2(油相仅有TTO、水相由全部药物组成)及纳米剂3(无油相、水相正常载药);取耳念珠菌CBS12766单克隆菌落制备耳念珠菌菌悬液,采用微量稀释法和平板涂布法检测3种中药纳米乳剂对耳念珠菌的最小抑菌浓度(MIC)和最小杀菌浓度(MBC);40只BALB/c小鼠尾静脉注射5 × 1010CFU/L菌悬液制作深部感染模型后均分为阳性对照组、异常行为1组、异常行为2组及异常行为3组,3种中药纳米乳化剂分别按照20µg/g剂量每天1次腹腔注射3 d,观察各组小鼠21 d内异常行为的发生情况;20只Balb/c小鼠随机分为阳性对照组、纳米剂1组、纳米剂2组、纳米剂3组尾静脉注射5 × 1010CFU/L菌悬液造模,3种中药纳米乳化剂分别按照20 µg/g剂量每天1次腹腔注射3 d,取肝、肾、心脏及脾脏匀浆检测载菌量、白细胞介素-6(IL-6)及IL-1β水平.结果 3种中药纳米乳剂对耳念珠菌的MIC与MBC均具有体外抑菌作用;小鼠异常行为曲线表明,与阳性对照组相比,3种中药纳米乳剂能够缓解小鼠病理状态(P＜0.05);与阳性对照组相比,纳米乳剂1～3组小鼠靶器官载菌量均降低(P＜0.05);与纳米剂2、3组相比,纳米剂1组杀菌效果更好(P＜0.05);与阳性对照组相比,小鼠靶器官促炎因子IL-6、IL-1 β表达均下降(P＜0.05);与纳米剂2、3组对比,纳米剂1促炎因子释放量显著下降(P＜0.05).结论 3种中药纳米乳剂对耳念珠菌深部感染均具有较好的抗菌效果,且纳米剂1效果比2、3更好.
Abstract
Objective To investigate the effects of three types of traditional Chinese medicine nanoemulsions on mice with deep-seated Candida auris(C.auris)infection.Methods Tea tree essential oil(TTO),Blumea balsamifera oil(BBO),Eucommia ulmoides,Paeonia lactiflora,Poria cocos,Angelica dahurica,and citric acid were used.High-pressure homogenization and ultrasonic testing were used to prepare nanoemulsions:Nanoagent 1(TTO,BBO,and all drugs in the aqueous phase),Nanoagents 2(TTO in the oil phase and all drugs in the aqueous phase)and Nanoagent 3(no oil phase and normal drug loading in the aqueous phase).Monoclonal Colony of Ear Candida CBS12766 was used to prepare ear C.auris Suspension.Microdilution method and plate coating method were applied to detect the minimum inhibitory concentration(MIC)and minimum bactericidal concentration(MBC)of three types of nanoagents against C.auris.After 40 BALB/c mice were injected 5 × 1010 CFU/L bacterial suspension via mouse tail vein to establish a deep-seated infection model,these mice were evenly divided into a positive control group,abnormal behavior group 1,abnormal behavior group 2 and abnormal behavior group 3.Three types of Nanoagents were intraperitoneally injected into the mice once a day at a dose of 20 μg/g for 3 days,and the occurrence of abnormal behavior in each group of mice was observed within 21 days.Twenty Balb/c mice were randomly divided into a positive control group,Nanoagent 1,Nanoagent 2,and Nanoagent 3 groups.A 5 × 105 CFU/L bacterial suspension was injected via mouse tail vein to generate a model.Three types of Nanoagents were intraperitoneally injected into the mice once a day at a dose of 20 μg/g for 3 d.Bacterial load,interleukin-6(IL-6),and IL-1β were measured in the homogenates of liver,kidney,heart,and spleen.Results Three types of Nanoagents exerted in vitro antibacterial effects on C.auris.The abnormal behavior curves of the mice showed that three types of Nanoagents could alleviate the pathological state of mice when compared to the positive control group(P＜0.05).When compared to the positive control group,the bacterial loads in the target organs of mice in groups 1-3 of Nanoagents were decreased(P＜0.05).When compared to groups 2 and 3 of Nanoagents,group 1 of Nanoagent had a better bactericidal effect(P＜0.05).When compared to the positive control group,the expressions of pro-inflammatory cytokines IL-6 and IL-1β in the target organs of the mice were decreased(P＜0.05).When compared to groups 2 and 3 of Nanoagents,the release of pro-inflammatory factors by Nanoagent 1 was significantly decreased(P＜0.05).Conclusion Three types of Nanoagents have good antibacterial effects on deep-seated C.auris infection.Nanoagent 1 is more effective than 2 and 3.
关键词
念珠菌属/感染/耳念珠菌/中药纳米乳剂/微量稀释法/促炎因子/最小抑菌浓度/最小杀菌浓度Key words
Candida/infection/Candida auris/nanoemulsion of Chinese medicine/microdilution method/pro-inflammatory cytokines/minimal inhibitory concentration/minimal bactericide concentra-tion引用本文复制引用
基金项目
国家自然科学基金(32060034)
西南喀斯特区域病原微生物防治学科创新引智基地(D20009)
贵州省高层次创新型人才项目(GCC[2022]036-1)
贵州省科技计划项目(黔科合基础ZK[2023]一般332)
出版年
2024
NETL
NSTL
万方数据