摘要
目的探讨洋葱中黄酮类化合物对结核分支杆菌的抑菌作用。方法选用56株结核分枝杆菌临床分离株作为研究对象,结核分枝杆菌标准菌株(H37RV)为对照用菌株,通过分格平板琼脂比例法抑菌试验,观察不同浓度洋葱中黄酮类化合物、黄酮与一线抗结核药物联合使用对结核分支杆菌的抑菌作用;小鼠腹腔巨噬细胞与结核分支杆菌共同孵育后,将细胞分为对照组、560μg/ml黄酮组、280μg/ml黄酮组、140μg/ml黄酮组、28μg/ml黄酮组、异烟肼(INH)组,给予相应药物干预,通过实时荧光定量PCR法检测巨噬细胞吞噬的结核分支杆菌DNA水平,通过ELISA法检测细胞培养液中细胞因子水平。结果与2. 0μg/ml INH、1. 0μg/ml利福平(REP)、5. 0μg/ml乙胺丁醇(EMB)、2. 0μg/ml链霉素(SM)比较,28μg/ml黄酮敏感率较低,560μg/ml、280μg/ml黄酮敏感率较高,差异有统计学意义(P <0. 05),560μg/ml黄酮及280μg/ml黄酮敏感率无明显差异(P> 0. 05)。与2. 0μg/ml INH、1. 0μg/ml REP、5. 0μg/ml EMB、2. 0μg/ml SM比较,各药物联合使用280μg/ml黄酮后敏感率均较高,差异有统计学意义(P <0. 05)。与INH组比较,280μg/ml黄酮组、560μg/ml黄酮组巨噬细胞吞噬的结核分支杆菌DNA扩增Ct值及拷贝数较高,28μg/ml黄酮组巨噬细胞吞噬的结核分支杆菌DNA扩增Ct值及拷贝数较低,差异有统计学意义(P <0. 05)。与INH组比较,280μg/ml黄酮组、560μg/ml黄酮组细胞培养液中水平IFN-γ、IL-1β水平较高,28μg/ml黄酮组细胞培养液中水平IFN-γ、IL-1β、IL-6水平较低,差异有统计学意义(P <0. 05)。结论洋葱中黄酮类化合物对结核分支杆菌有明显的体外抑菌作用,能够协同INH、REP、EMB及SM抑菌,还能促进巨噬细胞释放IFN-γ、IL-1β等细胞因子,提高巨噬细胞吞噬结核分枝杆菌的能力,在≤280μg/ml范围内,黄酮类化合物浓度越高作用越强。
Objective To investigate the inhibitory effects of flavonoids from onion on mycobacterium tuberculosis. Methods A total of 56 strains of mycobacterium tuberculosis isolated in clinical practice were selected as research objects,at the same time, mycobacterium tuberculosis standard strain （H37RV） was served as control strain.The agar plate agar method was used to observe the inhibitory effects of different concentrations of flavonoids from onion,flavonoids combined with first-line anti tuberculosis drugs on mycobacterium tuberculosis.Moreover after co incubation of mouse peritoneal macrophages with mycobacterium tuberculosis,the cells were divided into control group,560μg/ml flavone group,280μg/ml flavone group,140μg/ml flavone group,28μg/ml flavone group and INH group,receiving the corresponding drug intervention,then DNA levels of mycobacterium tuberculosis in macrophages were detected by real-time fluorescent quantitative PCR,and cytokine levels in cell culture medium were detected by ELISA. Results As compared with that of 2.0μg/ml isoniazid （INH）, 1.0μg/ml rifampicin （REP）,5.0μg/ml ethambutol （EMB） and 2.0μg/ml streptomycin （SM）,the sensitive rate of 28μg/ml flavonoids was lower,the sensitive rate of 560μg/ml flavonoids and 280μg/ml flavonoids were higher,there were significant differences （ P 〈0.05）. However there was significant difference in sensitive rate between 560μg/ml flavonoids group and 280μg/ml flavonoids group （ P 〉0.05）. As compared with that of 2.0μg/ml INH,1.0μg/ml REP,5.0μg/ml EMB, 2.0μg/ml SM,the sensitivity rate of every drug congbined with 280 μg/ml flavonoids was higher,there was significant difference （ P 〈0.05）.The amplified DNA Ct value and copy number of mycobacterium tuberculosis engulfed by macrophage in 280μg/ml flavone group and 560μg/ml flavone group were significantly higher than those in INH group （ P 〈0.05）,however,which in 28μg/ml flavone group were significantly lower than those in INH group （ P 〈0.05）.
作者
方草
FANG Cao(Department of Pharmacy,The Second Hospital Affiliated to Hainan Medical College,Hainan,Haikou 570311,China)
出处
《河北医药》
CAS
2018年第20期3045-3048,3053,共5页
Hebei Medical Journal
基金
海南省自然科学基金项目(编号:813242)
