目的 采用G蛋白偶联受体及酶活检测实验方法，探究痹祺胶囊对类风湿性关节炎关键靶点的调节作用，明确其作用机制。方法 选取与抑制滑膜炎症相关的靶点[环氧化酶-2（cyclooxygenase-2，COX-2）、一氧化氮合酶（nitric oxide synthase，NOS）、核因子-κB（nuclear factor-κB，NF-κB）、趋化因子受体4（C-C chemokine receptor type 4，CCR4）]，与抑制血管翳生成相关的靶点[血管内皮生长因子受体（vascular endothelial growth factor receptor 2，VEGFR2/KDR）]，与抑制基质降解相关的受体[基质金属蛋白酶-3（matrix metalloproteinase-3，MMP-3）]，以及与活血相关的靶点[凝血酶、磷酸二酯酶（cGMP-inhibited 3′,5′-cyclic phosphodiesterase，PDE3A）、肾上腺素受体α1（alpha-1A adrenergic receptor，ADRA1A）]为研究载体，通过胞内钙离子荧光和酶抑制剂检测技术评价痹祺胶囊中19个主要单体成分对受体的拮抗作用以及对酶的抑制活性。结果 19个化合物中，士的宁、党参炔苷、茯苓酸、丹参素、丹参酮II_A、丹酚酸B、迷迭香酸、人参皂苷Rb₁、阿魏酸、藁本内酯、牛膝皂苷D可显著抑制COX-2活性；马钱子碱、士的宁、党参炔苷、白术内酯III、茯苓酸、丹参素、丹参酮II_A、丹酚酸B、迷迭香酸、三七皂苷R₁、人参皂苷Rg₁、人参皂苷Rb₁、藁本内酯、蜕皮甾酮、甘草苷可显著抑制NOS活性；丹参素、迷迭香酸、丹酚酸B对NF-κB有较好拮抗作用；藁本内酯、牛膝皂苷IV、甘草次酸可拮抗CCR4；茯苓酸、牛膝皂苷IV、丹酚酸B、藁本内酯、甘草次酸能抑制VEGFR2/KDR；党参炔苷、丹参素、丹酚酸B、迷迭香酸、藁本内酯可抑制MMP-3活性；丹参素、丹参酮II_A、阿魏酸可抑制凝血酶活性，迷迭香酸、丹酚酸B、藁本内酯、甘草苷可抑制PDE3A活性；马钱子碱、士的宁、藁本内酯、牛膝皂苷IV、甘草次酸可拮抗ADRA1A。结论 痹祺胶囊可能通过干预COX-2、NOS、NF-κB、CCR4、VEGFR2/KDR、MMP-3、凝血酶、PDE3A及ADRA1A等关键靶点，发挥抗炎止痛、抑制血管翳形成及基质降解、活血作用，进而起到治疗类风湿性关节炎的药效作用。其药效物质基础可能为马钱子碱、士的宁、党参炔苷、白术内酯III、茯苓酸、丹参素、迷迭香酸、阿魏酸、丹参酮II_A、丹酚酸B、三七皂苷R₁、人参皂苷Rg₁、人参皂苷Rb₁、藁本内酯、蜕皮甾酮、牛膝皂苷D、甘草次酸、甘草苷。

Objective To explore the regulatory effect of Biqi Capsule (痹祺胶囊) on key targets of rheumatoid arthritis by G protein coupled receptor (GPCR) and enzyme activity assay. Methods The receptors cyclooxygenase-2 (COX-2), nitric oxide synthase (NOS), nuclear factor-κB (NF-κB) and C-C chemokine receptor type 4 (CCR4) related to the inhibition of synovial inflammation, vascular endothelial growth factor receptor 2 (VEGFR2/KDR) associated with the inhibition of pannus formation, matrix metalloproteinase-3 (MMP-3) related to the inhibition of matrix degradation, thrombin, cGMP-inhibited 3′,5′-cyclic phosphodiesterase (PDE3A), alpha-1A adrenergic receptor (ADRA1A) related to blood circulation were selected as research vectors. Intracellular calcium fluorescence detection and enzyme inhibitor detection techniques were used to evaluate the antagonistic effect of 19 main monomer components in Biqi Capsule on receptor and the inhibitory activity on enzyme. Results Among the 19 compounds, strychnine, lobetyolin, pachymic acid, danshensu, tanshinone IIA, salvianolic acid B, rosmarinic acid, ginsenoside Rb1, ferulic acid, ligustilide, and achyranthes saponin D significantly inhibited COX-2 activity; Brucine, strychnine, lobetyolin, atractylenolide III, pachymic acid, danshensu, tanshinone II_A, salvianolic acid B, rosmarinic acid, notoginsenoside R₁, ginsenoside Rg₁, ginsenoside Rb₁, ligustilide, ecdysterone and liquiritin could significantly inhibit NOS activity; Danshensu, rosmarinic acid and salvianolic acid B had good antagonistic effects on NF-κB; Ligustilide, achyranthes saponin IV and glycyrrhetinic acid could antagonize CCR4; Pachymic acid, achyranthes saponin IV, salvianolic acid B, ligustilide and glycyrrhetinic acid could inhibit VEGFR2/KDR; Lobetyolin, danshensu, salvianolic acid B, rosmarinic acid and ligustilide could inhibit MMP-3 activity; Danshensu, tanshinone II_A and ferulic acid could inhibit the activity of thrombin; Rosmarinic acid, salvianolic acid B, ligustilide and liquiritin could inhibit PDE3A activity; Brucine, strychnine, ligustilide, achyranthes saponin IV and glycyrrhetinic acid could antagonize ADRA1A. Conclusion Biqi Capsule may exert anti-inflammatory, analgesic, blood activating, inhibiting pannus formation, and matrix degradation effects by interfering with key targets of COX-2, NOS, NF-κB, CCR4, VEGFR2/KDR, MMP-3, thrombin, PDE3A, ADRA1A, and thus play a role in treatment of rheumatoid arthritis. Its pharmacodynamic material basis may be brucine, strychnine, lobetyolin, atractylenolide III, pachymic acid, danshensu, rosmarinic acid, ferulic acid, tanshinone II_A, salvianolic acid B, notoginsenoside R₁, ginsenoside Rg₁, ginsenoside Rb₁, ligustilide, ecdysterone, achyranthes saponin D, glycyrrhetinic acid and liquiritin.

国家自然科学基金重点项目（U21A20406）