Kang Byoung-Man, An Byoung-Kwan, Jung Won-Seok, Jung Ho-Kyung, Cho Jung-Hee, Cho Hyun-Woo, Jang Se Ji, Yun Young Beom, Kuk Yong In
Division of Traditional Korean Medicine Resource, National Development Institute of Korean Medicine, Jangheung 59338, Republic of Korea.
Department of Development in Oriental Medicine Resources, College of Life Science and Natural Resources, Sunchon National University, Sunchon 57922, Republic of Korea.
Int J Mol Med. 2016 Nov;38(5):1614-1620. doi: 10.3892/ijmm.2016.2765. Epub 2016 Sep 30.
The aim of this study was to identify major anti-inflammatory compounds in Alopecurus aequalis Sobol. (A. aequalis). The ethanol extract and the hexane-, dichloromethane-, ethyl acetate- and n-butanol-soluble fractions derived from A. aequalis were evaluated in order to determine their inhibitory effects on nitric oxide (NO) production in RAW 264.7 cells stimulated with lipopolysaccharide (LPS). The ethanol extract decreased NO production in a dose-dependent manner without any evidence of cytotoxicity at a concentration range of 0-200 µg/ml. The ethyl acetate soluble fraction was the most potent among the four soluble fractions. A compound was isolated by reversed-phase high-performance liquid chromatography from the ethyl acetate soluble fraction and this was identified to be tricin. Tricin inhibited the LPS-induced NO production in a dose-dependent manner without any evidence of cytotoxity at a concentration range of 1-100 µg/ml. Tricin also inhibited the LPS-induced production of prostaglandin E2. Western blot analysis indicated that tricin decreased the LPS-induced increase in the protein levels of inducible NO synthase and cyclooxygenase. In addition, tricin suppressed the production of intracellular reactive oxygen species in the LPS-stimulated RAW 264.7 cells, as measured by flow cytometry. Taken together, our results clearly indicate that tricin is a major functional anti-inflammatory compound which can be isolated from A. aequalis extracts.
本研究的目的是鉴定日本看麦娘中的主要抗炎化合物。对日本看麦娘的乙醇提取物以及衍生自日本看麦娘的己烷、二氯甲烷、乙酸乙酯和正丁醇可溶级分进行了评估，以确定它们对脂多糖（LPS）刺激的RAW 264.7细胞中一氧化氮（NO）产生的抑制作用。乙醇提取物在0 - 200μg/ml浓度范围内以剂量依赖方式降低NO产生，且无任何细胞毒性迹象。乙酸乙酯可溶级分在这四个可溶级分中活性最强。通过反相高效液相色谱法从乙酸乙酯可溶级分中分离出一种化合物，经鉴定为小麦黄素。小麦黄素在1 - 100μg/ml浓度范围内以剂量依赖方式抑制LPS诱导的NO产生，且无任何细胞毒性迹象。小麦黄素还抑制LPS诱导的前列腺素E2的产生。蛋白质印迹分析表明，小麦黄素降低了LPS诱导的诱导型一氧化氮合酶和环氧化酶蛋白水平的升高。此外，通过流式细胞术检测，小麦黄素抑制了LPS刺激的RAW 264.7细胞中细胞内活性氧的产生。综上所述，我们的结果清楚地表明，小麦黄素是一种可从日本看麦娘提取物中分离出的主要功能性抗炎化合物。
