Anti-inflammatory and antioxidant activities of constituents isolated from Pueraria lobata roots
Anti-inflammatory and antioxidant activities of constituents isolated from Pueraria lobata roots
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DOI：
10.1007/s12272-012-0508-x
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发表时间：
2012-05-01
影响因子：
6.7
通讯作者：
Choi, Jae Sue
中科院分区：
文献类型：
--
作者：
Jin, Seong Eun;Son, You Kyung;Choi, Jae Sue
为了评价葛根及其活性成分的抗炎和抗氧化活性，体外抑制RAW 264.7细胞中脂多糖（LPS）诱导的一氧化氮（NO）产生、诱导型一氧化氮合酶（iNOS）、环氧化酶-2（考克斯-2）蛋白表达和叔丁基过氧化氢（t-BHP）诱导的活性氧（ROS）产生的活性，测定了其对1，1-二苯基-2-苦基肼（DPPH）、过氧亚硝酸根（ONOO-）、一氧化氮（NO中心点）、超氧阴离子（中心点O-2（-））和总活性氧（ROS）的体外清除活性，以及对ONOO-介导的酪氨酸硝化的抑制活性。重复进行柱色谱，从抗炎和抗氧化剂EtOAc级分中分离出四种已知化合物：大豆苷元;染料木黄酮;葛根素;（+）-葛根素B-2-O-吡喃葡萄糖苷;从抗炎正己烷级分中分离出四种已知化合物：羽扇豆酮;羽扇豆醇;葛根素;香豆雌酚;从抗氧化剂n-BuOH级分中分离出七种已知化合物：尿囊素; 3 ′-羟基葛根素;大豆苷元8-C-芹菜糖基-（1 -> 6）-葡萄糖苷;葛根素;染料木苷; 3 '-甲氧基葛根素;大豆苷。在这些化合物中，羽扇豆酮和羽扇豆醇降低了LPS刺激的RAW 264.7细胞中的NO产生以及iNOS和考克斯-2蛋白水平。此外，羽扇豆醇对t-BHP引起的细胞内ROS生成具有显著的抑制活性。同时，3 '-羟基葛根素具有明显的ONOO-、NO中心点、总ROS清除活性和较弱的中心点O-2（-）清除活性，而3'-甲氧基葛根素具有ONOO-清除活性和较弱的NO中心点、O-2（-）清除活性，表明葛根素中3 '-羟基的存在对ONOO-、NO中心点的清除起重要作用，和总活性氧，以及抑制ONOO-介导的酪氨酸硝化机制。这些结果表明，野葛根及其成分可能是一种有用的治疗和预防各种炎症性疾病和氧化应激相关疾病的方法。
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 center dot), superoxide anion (center dot 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 center dot, total ROS scavenging activities, and weak center dot O-2 (-) scavenging activity, while 3'-methoxypuerarin showed ONOO- scavenging activity and weak NO center dot 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 center dot, 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.