黄连提取物抑制结核分枝杆菌的机理初探  

Preliminary study on inhibition mechanism of Coptis extract against Mycobacterium tuberculosis

作　　者：郎俊英 李桂梅 孙鹏[2] 杨丽敏[2] 张美仁 马晓辰 李斌[3] LANG Jun-ying;LI Gui-mei;SUN Peng;YANG Li-min;ZHANG Mei-ren;MA Xiao-chen;LI Bin;无(Hohhot Second Hospital,Hohhot,Inner Mongolia 010020,China;Inner Mongolia Medical University,Hohhot,Inner Mongolia 010051,China;The First Hospital of Hohhot City,Hohhot,Inner Mongolia 010031,China;Xing'an Vocational And Technical College,Ulanhot,Inner Mongolia 137400,China;Hohhot Occupational Disease Hospital,Hohhot,Inner Mongolia 010020,China)

机构地区：[1]呼和浩特市第二医院,内蒙古呼和浩特010020 [2]内蒙古医科大学,内蒙古呼和浩特010051 [3]呼和浩特市第一医院,内蒙古呼和浩特010031 [4]兴安职业技术学院,内蒙古乌兰浩特137400 [5]呼和浩特市职业病医院,内蒙古呼和浩特010020

出　　处：《中国热带医学》2022年第2期123-128,共6页China Tropical Medicine

基　　金：内蒙古自治区应用技术研究与开发资金计划项目(No.201702123)。

摘　　要：目的研究黄连提取物在体内及体外对结核分枝杆菌的抑制作用及部分机理。方法本实验用体外抑菌试验和体内造模实验检测黄连提取物抑制结核分枝杆菌(H37Ra)的作用及机理。抑菌试验:将黄连提取物按不同浓度添加到分枝杆菌固体培养基中,在培养基表面滴加H37Ra菌液,设空白组、利福平组做为对照,37℃培养2个月,观察有无菌落生长。造模实验:用结核分枝杆菌减毒株H37Ra感染小鼠,分为未感染组(正常组)、感染未治疗组(模型组)、感染治疗组(黄连组、利福平组)。做结核菌素试验验证造模阳性率,用ELISA实验检测小鼠血清IL-2的表达,使用RNA-seq检测小鼠肺的基因型改变。结果体外抑菌试验结果显示除空白组外其他各组均未见菌落生长,证明黄连提取物对结核菌有抑制作用。造模实验的结核菌素试验阳性率63%,证明造模成功。小鼠血清IL-2的表达显示黄连组IL-2的表达升高。小鼠肺RNA-seq检测测得数百条有意义的差异基因,揭示了黄连提取物抑制结核杆菌的作用机制,如Vipr2提高小鼠的细胞免疫,H2-Ea增强小鼠抵抗肺纤维化等。结论黄连提取物对结核分枝杆菌有抑制作用且抑制机理是多渠道、多靶点的。Objective To study the inhibitory effect of Coptis extract on Mycobacterium tuberculosis in vivo and in vitro,and its partial mechanism.Methods The inhibitory effect and mechanism of Coptis extract against Mycobacterium tuberculosis(H37Ra)were detected by Bacteriostatic test in vitro and modeling test in vivo.Bacteriostatic test:Coptis extract was added to the solid medium of Mycobacterium according to different concentrations,and H37Ra bacterial solution was dropped on the surface of the medium.The blank group and rifampicin group were set as the control group.Culture at 37℃ for two months,and the colony was observed.Model experiment:Mice were infected with Mycobacterium tuberculosis attenuated strain H37Ra and divided into uninfected group(normal group),infected untreated group(model group)and infected treatment group(Coptis extract group and rifampicin group).Tuberculin test was performed to verify the positive rate of model making,the expression of IL-2 in serum was detected by ELISA,and the genotype change of lung was detected by RNA-seq.Results In vitro bacteriostatic test showed that no colony growth was observed in all groups except blank group,which proved that Coptis extract had inhibitory effect on tuberculosis bacteria.The positive rate of tuberculin test in modeling experiment was 63%,which proved that modeling was successful.The expression of IL-2 in mouse serum showed that increased in the coptis group.Hundreds of significant differentially expressed genes were detected by RNA-seq assay in the lung of mice,revealing the mechanism of the coptis extract against M.tuberculosis,such as Vipr2 enhancing cellular immunity in mice,h2-Ea enhancing resistance to pulmonary fibrosis in mice,etc.Conclusion Coptidis extract has an inhibitory effect on M.tuberculosis and the inhibitory mechanism is multi-channel and multi-target.

关 键 词：黄连提取物 H37RA RNA-SEQ Vipr2 H2-Ea 

分 类 号：R378.911[医药卫生—病原生物学] R282.6[医药卫生—基础医学]