Purification of Anti-inflammatory Constituents from Anisomeles indica

• Main Authors: Shih-Chuan Hsieh, 謝世川
• Other Authors: Yew-Min Tzeng
• Format: Others
• Language: zh-TW
• Published: 2007
• Online Access: http://ndltd.ncl.edu.tw/handle/65041665945824206382

碩士 === 朝陽科技大學 === 生物技術研究所 === 95 === Anisomeles indica O. Kuntze (Labiatae) is an annual and biennial herb. In an alternative medicine this plant has been used as damp heat, carminative, stomachic, detoxification, pain relief, rheumatoid arthritis, gastroenteritis and other inflammatory diseases. In the present study, plant morphology and microscopic characteristics are appraising to confirm A. indica species. The whole plants of A. indica were extracted with methanol under reflux. The combined methanol extract was portioned with chloroform and n-butanol. Each layer was concentrated under reduced pressure individually, and then subjected to silica gel column chromatography, thin-layer chromatography, and semi-Preparative HPLC. Twenty-three pure compounds were isolated among 12 compounds structures were also determined. From chloroform layer, three steroids namely：β-sitosterol、stigmasterol、β-sitosterol-3-O-β-D-glucoside were obtained. The n-butanol layer yielded six phenylpropanoids namely,: betonyoside A、acetoside、isoacetoside、calceolarioside A、compneoside II、anomeric cistanoside F and two flavonoids:apigenin 7-O-(6"-O-p-coumaroylglucoside)、apigenin 7-O-β-D-glucuronopyranoside, and one phenolic acid : 3, 4-dihydroxybenzoic acid. Now studies are under progress on these isolates for their anti-inflammatory properties. Further, the aqueous and methanol extracts of whole plants, leaves, flowers and stems; and chloroform and n-butanol fractions of methanol extract, from A. indica were investigated for their anti-inflammatory activity on murine peritoneal macrophages. Treatment with A. indica extracts did not reduce cell viability at any dose used. However, all the extracts significantly inhibited the enhanced production of NO radicals, and pro-inflammatory cytokines (TNF-α, and IL-12) induced by LPS/IFN-γ in a dose dependent manner. In NO production inhibitory assays, methanol extracts were more active than the same with water. For example, methanol extract of leaves (ML), flowers (MF), and its CHCl3 fraction (MC) inhibited NO production by 90, 80 and 98%, respectively, at 20 μg/ml concentration. In the cytokines inhibitory assay, the methanol extracts ML, MF, and MC were the most active and inhibited TNF-α by 86, 77, and 92 and IL-12 by 88, 75, and 89%, respectively, at 20 μg/ml concentration.