过山枫中扁蒴藤素含量测定及其抑制HFLS-RA活性和迁移作用  

Content Determination of Pristimerin in Celastrus aculeatus Merr. and Its Inhibitory Effect on Viability and Migration of HFLS-RA

作　　者：邓秋狄[1] 白殊同[1] 高皖皎 佟丽[1] 

机构地区：[1]南方医科大学中医药学院,广东广州510515

出　　处：《中药新药与临床药理》2015年第6期818-821,共4页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金项目(31240045)

摘　　要：目的建立高效液相色谱法（HPLC）测定过山枫药材中扁蒴藤素含量的方法,研究扁蒴藤素对人类风湿关节炎滑膜成纤维细胞（HFLS-RA）活性和迁移能力的影响。方法采用C18色谱柱（4.6 mm×250 mm,5μm）,甲醇-0.1%磷酸水为流动相进行梯度洗脱,流速1.40 m L·min-1,柱温30℃,检测波长424 nm;MTT法测定HFLS-RA的活性;划痕实验观察扁蒴藤素对HFLS-RA迁移的影响。结果扁蒴藤素在浓度为0.08~0.64g·L-1呈良好的线性关系（r=0.9995）,平均回收率为101.09%,RSD=2.58%;浓度为0.50~1.0μmol·L-1的扁蒴藤素,抑制HFLS-RA活性具有剂量和时间的依赖性关系,能够明显减少血管内皮生长因子（VEGF）诱导HFLS-RA的迁移。结论所建立的扁蒴藤素含量测定方法简便、准确,可用于过山枫药材质量的控制;扁蒴藤素具有抑制HFLS-RA活性和迁移的作用。Objective To establish an HPLC method for the content determination of pristimerin in Celastrus aculeatus Merr. and to study the effect of pristimerin on the viability and migration of human fibroblast-like synoviocytes- rheumatoid arthritis（HFLS-RA）. Methods The determination of pristimerin was carried out on C 18 column（4.6 mm× 250 mm, 5 μm） with the mobile phase consisting of methanol-0.1% phosphate acid by gradient elution at a flow rate of 1.40 mL.min-1. The detection wavelength was set at 424 nm and the column temperature was maintained at 30 ℃. The viability of HFLS-RA was quantified by MTT assay, and the migration ability of HFLS-RA was detected by wound-healing migration test. Results Good linearity of pristimerin was obtained in the range of 0.08-0.64 g.L-1（r=0.9995）. The average recovery was 101.09 %, and RSD was 2.58 %. In concentration range of 0.5 to 1.0 μmol.L-1, pristimerin inhibited cell viability in a dose- and time-dependent manner, and obviously reduced HFLS-RA migration induced by vascular endothelial growth factor （VEGF）. Conclusion The method is simple and rapid, and could be used for evaluating the quality of Celastrus aculeatus Merr.. Pristimerin possesses certain inhibitory effects on viability and migration of HFLS-RA.

关 键 词：过山枫 扁蒴藤素 高效液相色谱法 HFLS-RA迁移 

分 类 号：R284.1[医药卫生—中药学]