Seven new naphthoquinone diglycosides (1-7), three new anthraquinones (8-10), and eight known analogues were obtained from the aerial parts of Mitracarpus hirtus collected from West Africa in a bioassay-guided phytochemical investigation. All isolated compounds were elucidated by comparison with the literature and interpretation of spectroscopic data, and the absolute configurations of the new naphthoquinone diglycosides (1-10) were confirmed by chemical methods and ECD calculations. Notably, compound 1 was found to be the first naphthoquinone diglycoside containing carboxylic acid and isopentenyl side chains isolated from a species in the genus Mitracarpus. Compounds 6-18 showed antibacterial activity against multiple Helicobacter pylori strains with MIC values ranging from 0.0625 to 64 μg/mL. Particularly, 1-hydroxybenzoisochromanquinone (17) and benzo[g]isoquinoline-5,10-dione (18), with MIC values of 0.0625 and 0.125 μg/mL, displayed 32-512-fold higher potencies than a positive control, metronidazole. Compound 18 also demonstrated high antibiofilm activity and killed biofilm-encased Helicobacter pylori cells more effectively than metronidazole.

Ethnopharmacological relevance: Lonicerae Japonicae Caulis, the dried stem and branch of Lonicera japonica Thunb., is a Chinese Materia Medica known as Ren Dong Teng in Chinese with long use history in the traditional Chinese medicine (TCM) prescriptions. Lonicerae Japonicae Caulis possesses heat-clearing and detoxifying functions according to the TCM theory. In recent years, a large amount of experimental and clinical studies proved good anti-inflammatory effects of some heat-clearing and detoxifying herbs. The present study aims to reveal the anti-inflammatory property and functional substances of Lonicerae Japonicae Caulis. Materials and methods: For anti-inflammatory activity test, LPS-induced RAW 264.7 macrophages, DSS-induced SPF male C57BL/6J mice model, and LPS-induced SPF male ICR mice model were used in vitro and in vivo, respectively. The behavioral changes, organ damage, and the expression of inflammatory factors such as TNT-alpha and IL-6 mRNA expression were measured for activity evaluation. Lonicerae Japonicae Caulis samples were prepared by solvent extraction and subsequent column chromatography. The main components were identified and determined using UPLC-UV analysis as well as NMR interpretation after purification. To testify the contribution of main components for the anti-inflammatory activity, different samples were also prepared by compound-knockout strategy. Results: Ethanol extract of Lonicerae Japonicae Caulis could attenuate sickness symptoms in mice such as diarrhea, less activity, and depression. It could also alleviate multiple organ damage, and significantly inhibit the expression of pro-inflammatory factors such as TNF-alpha, IL-1 beta, IL-6 and IFN-gamma in mice. Furthermore, the isochlorogenic acid-rich and biflavonoid-rich fractions and isochlorogenic acids A and C, and ochnaflavone could significantly down-regulate the mRNA expression of TNF-alpha and IL-6 in LPS-induced RAW 264.7 macrophages. Conclusions: Lonicerae Japonicae Caulis possesses anti-inflammatory property. Its isochlorogenic acid-rich and biflavonoid-rich fractions do the major contribution. And their main components, isochlorogenic acids A and C, and ochnaflavone, take main responsibility for the anti-inflammatory property.

目的 优选黄芩茎叶总黄酮制备工艺,为黄芩茎叶资源的合理利用开发提供依据.方法 以黄芩茎叶中8个黄酮类化合物的转移率为指标,采用单因素考察结合正交试验优选黄芩茎叶黄酮的提取工艺;比较酸沉法和大孔树脂法纯化茎叶总黄酮的转移率,并对大孔树脂纯化工艺进行优选.结果 以12倍量40％乙醇热回流提取3次,每次1 h,为最佳提取工艺.提取物的纯化结果显示,大孔树脂法中黄芩茎叶黄酮的总体转移率(92.53％)显著高于酸沉法(76.69％).在pH为1.01时,酸沉法虽然可以使野黄芩苷等化合物转移率达90％以上,但含量丰富的二氢黄酮类化合物异红花素-7-O-β-D-葡萄糖醛酸苷、红花素-7-O-β-D-葡萄糖醛酸苷的转移率仅为64.76％、68.57％,而使用大孔树脂法纯化,其转移率均可达90％以上,可以实现各目标成分的高效转移.经优选,0.26 g/mL药液上样,上样量为每10 g HP-20型干树脂上样2.08 g干燥提取物,树脂柱径高比为1:3,4 BV纯水除杂后,以30％乙醇洗脱4 BV,为最佳纯化工艺,所得总黄酮质量分数可达66.03％.结论 建立了稳定且环境友好型黄芩茎叶总黄酮制备工艺,可为黄芩茎叶的进一步开发利用提供参考.

为探讨黄芩茎叶、藤茶及两者联合应用对金黄色葡萄球菌和痢疾杆菌的抑制作用,研究采用微量肉汤稀释法分别测定黄芩茎叶和藤茶对金黄色葡萄球菌和痢疾杆菌的最小抑菌浓度(Minimal inhibit concentration,MIC)及时间-杀菌曲线;采用棋盘式微量稀释法测定两者联合使用后的FIC指数.结果 显示,黄芩茎叶和藤茶对金黄色葡萄球菌的MIC分别为1、0.25 mg/mL,对痢疾杆菌的MIC分别为4、0.50 mg/mL.两者联合使用,对金黄色葡萄球菌和痢疾杆菌的FIC指数分别为0.75、0.625,表现出相加的作用效果.研究提示基于联合使用,可进一步提高黄芩茎叶的抑菌效力,为实现黄芩茎叶的资源化利用提供科学依据.

目的:建立门氏护胃颗粒的薄层鉴别与含量测定研究方法.方法:采用薄层色谱对门氏护胃颗粒中人参、连翘、甘草、干姜进行定性鉴别,采用HPLC法对连翘酯苷A和连翘苷进行含量测定.结果:薄层色谱斑点分离效果良好,斑点显色清晰,且无干扰,重复性好.连翘酯苷A与连翘苷在0.61～6.1μg、0.246～2.46μg内线性关系良好(r=0.999):平均含量分别为1.00、0.37mg/g;平均回收率(n=6)分别为96.06％,97.09％.结论:主药薄层色谱定性鉴别特征性强,含量测定方法重现性好,操作简便,可作为门氏护胃颗粒的质量标准研究方法.