| Berberis agricola Ahrendt belongs to the genera Berberis, Berberidaceae family indigenous to Tibetan Autonomous Region. The stems and roots of Berberis agricola are commonly used as febrifuge, antiseptic, cathartic, diuretic, antidysenteric, antibilious. And they are also used for treatment of inflammations, such as faucitis, conjunctivitis, enteritis, mastitis. Phytochemical investigation of the ethanol extracts from the aerial part led to the isolation of 11 compounds by silica gel column chromatography and preparation thin layer chromatography. Their structures were identified by the spectroscopic analysis methods including IR, UV, EI-MS, 1H-NMR and 13C-NMR spectra. There were seven alkaloids, three organic acids and one phytosterol: berberine(1), columbamine(2), jatrorrhizine(3), berberrubine(4), berbamine(5), cetanic acid(6), tetracosanoic acid(7), bebamine-2'-β-N-oxide(8), guattegaumerine(9), 4-hydroxybenzoic acid(10),β-sitosterol (11). All of them were obtained for the first time from Berberis agricola Ahrendt.The antibacterial activities of berberine (1), columbamine (2) and jatrorrhizine (3) from Berberis agricola Ahrendt were studied. The minimum inhibitive concentrations (MIC) of the three alkaloids against 13 species of bacteria (Staphylococcus aureus, Klebsiella pneumoniae pneumoniae, nonfermenters, Pseudomonas aeruginosa, Serratia marcescens , Entembacer cloacae, Proteus mirabilis, Stenotorphomonas maltophilia, Enterococcus sp, Enterobacter aerogens , Proteus vuigars,β-hemolytic streptococcus, Corynebacterium pseudodiphtheriticum) were observed by tube dilution method. It was showed that compounds 1-3 displayed significant antibacterial activities against Staphylococcus aureus with the MIC 31.25, 62.5, 62.5μg/mL, respectively. Only columbamine showed antibacterial activities against Proteus mirabilis (MIC=500μg/ml) among these alkaloids. Berberine had a broader antibacterial spectrum than the other two. Berberine had no obviously antibacterial activities against Entembacer cloacae and Stenotorphomonas maltophilia with the MIC＞500μg/mL, but its MIC against the other 10 species of bacteria were between 31.25-500μg/mL. Stenotorphomonas maltophilia, Enterococcus sp,β-hemolytic streptococcus could be inhibited by jatrorrhizine with the MIC 250μg/mL.Compounds 1-7 were tested for in vitro cytotoxic bioassay. The result showed that compounds 5 and 7 exhibited cytotoxic activities. The median effective dose (EC50) of 5 against HCT8 and A549 were 8.34μg/mL and 7.89μg/mL, respectively, and 7 against A549 was 10.84μg/mL. The cytotoxic bioassay of other compounds is in progress. |
