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目的:对愈肾汤总黄酮的化学成分和活性进行研究,为处方的活性成分揭示和作用机制阐明奠定基础。方法:采用体外H2O2诱导人脐静脉内皮细胞(HUVEC)损伤实验评价愈肾汤总黄酮的活性。并采用硅胶柱、大孔吸附树脂、葡聚糖凝胶色谱和高效液相制备等方法分离纯化愈肾汤总黄酮的化学成分,根据理化性质和波谱数据鉴定其结构。结果:愈肾汤总黄酮具有明显的抗过氧化氢诱导的HUVEC损伤和下调PAI-1表达(P<0.05)的作用。从愈肾汤总黄酮部位分离得到9个黄酮类化合物分别为毛蕊异黄酮(1)、蒙花苷(2)、3’,4’-二羟基-7-O-β-D-葡萄糖苷(3)、3’-羟基-4’-甲氧基-7-O-β-D-葡萄糖苷(4)、槲皮素(5)、山柰酚(6)、芦丁(7)、红车轴草异黄酮-7-O-β-D-葡萄糖苷(8)、4’-羟基二氢黄酮-7-O-β-D-葡萄糖苷(9)。结论:愈肾汤总黄酮具有抗HUVEC氧化损伤作用,所分离的9个黄酮化合物均为首次从该方中分离得到,化合物1,3,4,8,9可能来自于黄芪,化合物2,5～7可能来自于小蓟和仙鹤草,其中3,4和9为首次从组方的单味药中分离得到。
OBJECTIVE: To study the chemical constituents and activities of the total flavonoids in Yushen Decoction and to lay the foundation for elucidation of the active ingredients and elucidation of the mechanism of action. Methods: HUVEC induced injury of human umbilical vein endothelial cells (HUVEC) in vitro was used to evaluate the total flavonoids activity of Yushen Tang. The chemical constituents of the total flavonoids of Yunshen Decoction were separated and purified by silica gel column, macroporous adsorption resin, Sephadex GLC and HPLC. The structures of the compounds were identified by their physical and chemical properties and spectral data. Results: The total flavones of Yu Shen Tang had significant anti-HUVEC-induced HUVEC injury and down-regulation of PAI-1 expression (P <0.05). Nine flavonoids were isolated from the total flavonoids fraction of Yushen Tang, which were as follows: calycosin (1), monatin (2), 3 ’, 4’-dihydroxy-7-O-β-D-glucoside ), 3’-hydroxy-4’-methoxy-7-O-β-D-glucoside (4), quercetin (5), kaempferol (6), rutin Grass isoflavone-7-O-β-D-glucoside (8), 4’-hydroxyflavone-7-O-β-D-glucoside (9). CONCLUSION: The total flavonoids of Yanshen Decoction has the anti-HUVEC oxidative damage effect. The nine flavonoids isolated from this extract were obtained from this compound for the first time. Compounds 1, 3, 4, 8 and 9 may be derived from Astragalus membranaceus, compounds 2 and 5 ~ 7 may come from the thistle and Agrimony, which 3,4 and 9 for the first time from the group of single herbs isolated.