Five kinds of coumarin compounds were successfully purified from Angelica gigas Nakai by using recyclingpreparative HPLC and identified as decursin (1), decursinol angelate (2), 7-demethylsuberosine (3), marmesin (4), and decursinol (5) by NMR analyses. None of the purified compounds in ethanol show...
Five kinds of coumarin compounds were successfully purified from Angelica gigas Nakai by using recyclingpreparative HPLC and identified as decursin (1), decursinol angelate (2), 7-demethylsuberosine (3), marmesin (4), and decursinol (5) by NMR analyses. None of the purified compounds in ethanol showed DPPH radical scavenging activity, while the A. gigas extract (AGNEX) displayed a significant level of activity. Interestingly, compounds 3 in phosphate buffered saline (PBS) showed good $ABTS^+$ radical scavenging activity ($IC_{50}=8.1{\mu}g$/mL) as did compounds 4 and 5. The anti-inflammatory activities of the purified compounds were evaluated and compared using the NO concentration assay and western blot analysis on LPS-stimulated RAW 264.7 cells. NO production was significantly suppressed by all the compounds in a dose-dependent manner among which compounds 1, 2, and 3 showed very good activities with $IC_{50}$ values of 7.4, 6.5, and $7.6{\mu}g$/mL, respectively. Treatment with compounds 1-5 effectively suppressed the expression levels of iNOS, IL-1$\beta$, and COX-2, which are responsible for promoting the inflammatory process. Thus, the ethanol extract and coumarin compounds of A. gigas Nakai hold promise for use as potential anti-inflammatory agents.
Five kinds of coumarin compounds were successfully purified from Angelica gigas Nakai by using recyclingpreparative HPLC and identified as decursin (1), decursinol angelate (2), 7-demethylsuberosine (3), marmesin (4), and decursinol (5) by NMR analyses. None of the purified compounds in ethanol showed DPPH radical scavenging activity, while the A. gigas extract (AGNEX) displayed a significant level of activity. Interestingly, compounds 3 in phosphate buffered saline (PBS) showed good $ABTS^+$ radical scavenging activity ($IC_{50}=8.1{\mu}g$/mL) as did compounds 4 and 5. The anti-inflammatory activities of the purified compounds were evaluated and compared using the NO concentration assay and western blot analysis on LPS-stimulated RAW 264.7 cells. NO production was significantly suppressed by all the compounds in a dose-dependent manner among which compounds 1, 2, and 3 showed very good activities with $IC_{50}$ values of 7.4, 6.5, and $7.6{\mu}g$/mL, respectively. Treatment with compounds 1-5 effectively suppressed the expression levels of iNOS, IL-1$\beta$, and COX-2, which are responsible for promoting the inflammatory process. Thus, the ethanol extract and coumarin compounds of A. gigas Nakai hold promise for use as potential anti-inflammatory agents.
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제안 방법
In this study, NO production was significantly suppressed by AGNEX and compounds 1～3 in a dose-dependent manner with an IC50 of 10.3, 7.4, 6.5, and 7.6 μg/mL, respectively (Fig. 4).
gigas Nakai using recycling preparative HPLC in a single step. In this study, five coumarins, including decursin and decursinol angelate, were successfully purified using the recycling mode from AGNEX without the need for conventional column chromatography (Fig. 1). First, decursin (1) and decursinol angelate (2) were purified from AGNEX using the recycling mode together with five fractions that eluted earlier.
In this study, five types of major coumarins were successfully isolated from A. gigas Nakai by a simple and rapid procedure that used recycling preparative chromatography. In addition, the anti-inflammatory activities of these compounds were compared and evaluated using a radical scavenging assay, NO concentration assay and western blot analyses of inducible nitric oxide synthase (iNOS), interleukin-1β (IL-1β) and Cyclooxygenase 2 (COX-2) protein expression in LPS-stimulated RAW 264.
The chemical structures of these components were confirmed by comparing the UV, 1H, 13C and 2D-NMR spectral data to published spectra. The structures of the compounds were shown in Fig.
대상 데이터
HPLCgrade acetonitrile (ACN) and water were from Burdick & Jackson (Morristown, NJ, USA). Ethanol (95%) was from DUKSAN (Ansan, Korea). Other chemicals were of analytical grade.
gigas Nakai was purchased from the local market of Korea. The murine macrophage RAW 264.7 cell line was obtained from American Type Culture Collection and cultured in DMEM supplemented with 10% FBS, 2 mM L-glutamine, and 2 mM penicillin.
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