高速逆流色谱法提纯盐酸黄连素对粪肠球菌抑菌效果的研究

Antibacterial effect study of berberine hydrochloride purified by high-speed counter-current chromatography on enterococcus faecalis

目的探讨高速逆流色谱法(HSCCC)提纯黄连中盐酸黄连素对粪肠球菌体外抑菌效果的影响。方法选择150株临床分离的粪肠球菌株为研究对象,随机分为Ⅰ组、Ⅱ组与Ⅲ组各50株,均配制成2000～3000cfu/ml的粪肠球菌液;将黄连制成中药煎剂,配制成浓度为1.0g/ml的A药液;将HSCCC提纯黄连中的盐酸黄连素配制成浓度为1.0g/ml的B药液;Ⅰ组将粪肠球菌液涂到用A药液与M-H培养基混合的培养基上,Ⅱ组将粪肠球菌液涂到用B药液与M-H培养基混合的药液培养基上,Ⅲ组将粪肠球菌液涂到M-H培养基上,上述培养基均采用环丙沙星(CIP)、红霉素(ERY)、四环素(TE)和青霉素(PEN)药敏纸观察培养基药敏情况。结果Ⅰ组中CIP、ERY、TE和PEN的耐药率分别为26.0%、40.0%、36.0%、20.0%;Ⅱ组中CIP、ERY、TE和PEN的耐药率分别为10.0%、26.0%、14.0%、8.0%;Ⅲ组中CIP、ERY、TE和PEN的耐药率分别为46.0%、80.0%、44.0%、26.0%。在对环丙沙星、红霉素、四环素和青霉素的耐药率方面,Ⅲ组>Ⅰ组>Ⅱ组,三组间耐药率差异均有统计学意义(P<0.05)。结论 HSCCC提纯黄连中盐酸黄连素对临床分离的粪肠球菌菌株具有较强的抑菌活性,其抑菌作用比黄连煎剂强。

Objective To explore the influence of bacteriostasis in vitro on enterococcus faecalis by berberine hy- drochloride purified from rhizoma coptidis by high-speed counter-current chromatography （HSCCC）. Methods 150 strains of wild enteroeoccus faecalis isolated from the clinic were selected and evenly divided into group Ⅰ ,group Ⅱ, and group Ⅲ in randomly.They were formulated as 2000-3000 cfu/ml enterococcus faecalis solution.After decocting, rhizoma coptidis was prepared as solution A with the concentration of 1.0 g/ml.The berberine hydrochloride purified by HSCCC was formulated as solution B with the concentration of 1.0 g/ml.The enterococcus faeealis solution was smeared on mixed culture medium of solution A and M-H in group Ⅰ ,mixed culture medium of solution B and M-H in group Ⅱ ,and merely on M-H medium in group Ⅲ respectively.The culture medium mentioned above were all past- ed with drug sensitivity paper of ciprofloxacin （CIP）,erythrocin （ERY）,tetraeycline （TE）,and penicillin （PEN） in order to observe the drug sensitivity of the culture medium. Results The drug resistance rates of CIP,ERY,TE,and PEN in group I were 26.0%,40.0%,36.0%,and 20.0% respectively;and it was 10.0%,26.0%,14.0% and 8.0% respectively in group Ⅱ ;in group Ⅲ was 46.0%,80.0%,44.0% and 26.0% in turn.In the aspect of drug resistance rates of CIP,ERY, TE and PEN,group Ⅲ 〉group Ⅰ 〉group Ⅱ ,and the differences of drug resistance rates in the three groups were all sta- tistically significant（P〈0.05）. Conclusion The berberine hydrochloride from rhizoma coptidis purified by HSCCC has a strong baeteriostafic activity on enterococcus faecalis isolated from the clinic,whose bacteriostatic action is stronger than that of rhizoma coptidis decoction.