大黄及黄柏提取物体外抗沙眼衣原体活性研究

Research on monomers activities of against chlamydia trachomatis in vitro extracted from rhubarb root and rhizome and amur cork-tree

目的:分别检测中药大黄、黄柏提取物即包括大黄酸、盐酸小檗碱等在内的16种中药单体体外抗沙眼衣原体(Ct)活性,探讨复杂中药构成中起主要作用的成分;研究具有抗沙眼衣原体活性的各单体间是否存在协同作用。方法:1应用微量Mc Coy细胞培养法分别检测大黄酸、盐酸小檗碱等在内的16种中药单体体外抗Ct的MIC值和MBC值;2运用棋盘格稀释法,将在初步检测中显示对Ct有抑制和杀灭作用的中药单体间分别进行两两组合,在96孔板上进行稀释,应用微量Mc Coy细胞培养法检测两中药单体联用的部分抑菌浓度(FIC)。结果:1大黄提取物芦荟大黄素、大黄素、大黄酸具有较强的抗Ct活性,其MIC与MBC相同,分别为2.44、4.89、19.54μg/m L;黄柏提取物盐酸小檗碱具有卓越的抗Ct活性,其MIC与MBC亦相同,为1.22μg/m L;2中药单体大黄酸与芦荟大黄素、大黄酸和大黄素联合使用时有协同抗Ct活性作用,FIC值分别为0.38、0.50;大黄素与芦荟大黄素、芦荟大黄素与盐酸小檗碱联合使用时既无拮抗,亦无协同,FIC值分别为0.53、1.03。结论:1大黄提取物芦荟大黄素、大黄素、大黄酸和黄柏提取物盐酸小檗碱4种中药单体均具有较强的体外的抗Ct活性,优于原药水提液;2大黄的3种不同提取物大黄酸与芦荟大黄素、大黄酸与大黄素联合使用时有协同抗Ct作用,为大黄抗Ct活性提供了药理基础,为临床使用大黄治疗Ct感染提供了药理依据。

Objective：①To test the activities of against chlamydia trachomatis （Ct） of 16 extracts in rhubarb root and rhizome and amur cork-tree which with rheum acid and berberine hydrochloride included, and investigate the main active components in complex constituents of traditional Chinese medicine （TCM）. ②To research the synergistic action among various monomers with activities of against Ct. Methods： ①The minimal inhibitory concentration （MIC） and minimal bactericidal concentration （MBC） of 16 monomers with activities of against Ct were tested through culturing McCoy cell in vitro by using micro-culture technique. ②Combination of two monomers with inhibition and killing effect on Ct were diluted by using checkerboard dilution method in 96-well plates, and the fractional inhibitory concentration （FIC） of the combination of monomers was determined through culturing McCoy cell in vitro by using micro-culture technique. Results： ①he activities of against Ct of rheum acid, aloe-emodin and rheum emodin in rhubarb root and rhizome was fairly strong, and its MIC and MBC were both of 1.221ag/mL. ②There were synergistic effects of against Ct when combined use with aloe-emodin and rheum acid, rheum acid and rheum emodin, and the FIC values of the two combinations were 0.38 and 0.50 respectively. There was neither antagonism nor synergy when combined use with rheum emodin and aloe-emodin, aloe-emodin and berberine hydrochloride, and the FIC values of the two combinations were 0.53 and 1.03 respectively. Conclusion： ①The activities of against Ct of rheum acid, aloe- emodin and rheum emodin in rhubarb root and rhizome and berberine hydrochloride in amur cork-tree is fairly strong, which is superior to the water extract of original herbs, and worth to be further studied. ②There are synergistic effects of against Ct when combine use with aioe-emodin and rheum acid, rheum acid and rheum emodin extracted from rhubarb root and rhizome, which lay the pharmacological foundation for researches on against Ct activities of rhubarb root and rhizome and provides pharmacological basis for treating Ct infection with rhubarb root and rhizome in clinical.