Basic Herbal Medicine Research Group, Korea Institute of Oriental Medicine, Daejeon, Korea.
Arch Pharm Res. 2012 May;35(5):823-37. doi: 10.1007/s12272-012-0508-x. Epub 2012 May 29.
In order to evaluate the anti-inflammatory and antioxidant activities of Pueraria lobata roots and its active components, in vitro inhibitory activities against lipopolysaccharide (LPS)-induced nitric oxide (NO) production, inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) protein expression, and tert-butylhydroperoxide (t-BHP)-induced reactive oxygen species (ROS) generation in RAW 264.7 cells, as well as in vitro scavenging activities against 1,1-diphenyl-2-picrylhydrazyl (DPPH), peroxynitrite (ONOO(-)), nitric oxide (NO·), superoxide anion (·O(2)(-)) and total ROS, and inhibitory activities against ONOO(-)-mediated tyrosine nitration, were determined. Repeated column chromatography was performed to isolate four known compounds from the anti-inflammatory and antioxidant EtOAc fraction: daidzein; genistein; puerarin; (+)-puerarol B-2-O-glucopyranoside; four known compounds from the anti-inflammatory n-hexane fraction: lupenone; lupeol; puerarol; coumestrol; seven known compounds from the antioxidant n-BuOH fraction: allantoin; 3'-hydroxypuerarin; daidzein 8-C-apiosyl-(1→6)-glucoside; puerarin; genistin; 3'-methoxypuerarin; daidzin. Among these compounds, lupenone and lupeol reduced NO production, as well as iNOS and COX-2 protein levels in LPS-stimulated RAW 264.7 cells. Furthermore, lupeol showed significant inhibitory activity against intracellular ROS generation by t-BHP. Meanwhile, 3'-hydroxypuerarin showed marked ONOO(-), NO·, total ROS scavenging activities, and weak ·O(2)(-) scavenging activity, while 3'-methoxypuerarin showed ONOO(-) scavenging activity and weak NO· and O(2)(-) scavenging activities, suggesting that existence of the 3'-hydroxyl group in puerarin plays an important role in the scavenging of ONOO(-), NO·, and total ROS, as well as inhibiting the ONOO(-)-mediated tyrosine nitration mechanism. These results indicate that P. lobata roots and its constituents may be a useful therapeutic and preventive approach to various inflammatory diseases and oxidative stress-related disease.
为了评估葛根及其活性成分的抗炎和抗氧化活性，采用体外法测定了葛根乙酸乙酯提取物对脂多糖（LPS）诱导的一氧化氮（NO）生成、诱导型一氧化氮合酶（iNOS）、环氧化酶-2（COX-2）蛋白表达及叔丁基过氧化物（t-BHP）诱导的活性氧（ROS）生成的抑制作用，同时测定了葛根提取物对 1,1-二苯基-2-苦基肼（DPPH）、过氧亚硝酸盐（ONOO(-)）、一氧化氮（NO·）、超氧阴离子（·O(2)(-)）和总 ROS 的清除活性，以及对 ONOO(-)介导的酪氨酸硝化的抑制活性。采用反复柱层析法从抗炎和抗氧化 EtOAc 部分分离得到 4 种已知化合物：大豆苷元；染料木素；葛根素；(+)-葛根醇 B-2-O-吡喃葡萄糖苷；从抗炎 n-己烷部分分离得到 4 种已知化合物：羽扇豆酮；羽扇醇；葛根醇；黄豆黄素；从抗氧化正丁醇部分分离得到 7 种已知化合物：尿囊素；3'-羟基葛根素；大豆苷 8-C-阿拉伯糖苷（1→6）-葡萄糖苷；葛根素；金雀异黄素；3'-甲氧基葛根素；大豆苷。在这些化合物中，羽扇豆酮和羽扇醇降低了 LPS 刺激的 RAW 264.7 细胞中 NO 的产生以及 iNOS 和 COX-2 蛋白水平。此外，羽扇醇对 t-BHP 诱导的细胞内 ROS 生成具有显著的抑制作用。同时，3'-羟基葛根素表现出显著的 ONOO(-)、NO·、总 ROS 清除活性和较弱的·O(2)(-)清除活性，而 3'-甲氧基葛根素表现出 ONOO(-)清除活性和较弱的 NO·和 O(2)(-)清除活性，提示葛根中的 3'-羟基在清除 ONOO(-)、NO·和总 ROS 以及抑制 ONOO(-)介导的酪氨酸硝化机制中发挥重要作用。这些结果表明，葛根及其成分可能是治疗和预防各种炎症性疾病和氧化应激相关疾病的一种有效方法。
