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Jeong-Yong Park1, 2
, Ji Yeon Lee1, 2, Hyung Don Kim1, Gwi Yeong Jang1, Kyung Hye Seo1
, Ji Yeon Lee1, 2, Hyung Don Kim1, Gwi Yeong Jang1, Kyung Hye Seo1
Abstract
Purpose
Astragalus membranaceus (AM) is an important traditional medicinal herb. Pharmacological research has indicated that AM has various physiological activities such as antioxidant, anti-inflammatory, immunoregulatory, anticancer, hypolipidemic, antihyperglycemic, and hepatoprotective activities. The bioactive substances responsible for the physiological activities in AM, including many antioxidant substances, change during the roasting process. This study investigated and compared the changes in the antioxidant constituents of AM caused by roasting.
Methods
DPPH (1,1-diphenyl-2-picryl hydrazyl) and ABTS+ (2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt) radical scavenging activities and their total phenolic content (TPC) were measured. High-performance liquid chromatography (HPLC) analysis was performed to confirm any changes in the isoflavonoids of roasted AM (R-AM),. The cell viability of UVB-induced HDF (Human dermal fibroblast) cells treated with AM and R-AM extracts was investigated. The comet assay was used to examine the inhibitory effects of R-AM extracts on DNA damage caused by oxidative stress.
Results
The DPPH and ABTS+ radical scavenging activities were 564.6±20.9 and 108.2±3.1 (IC50 value) respectively, from the 2R-AM. The total phenol content was 47.80±1.40 mg GAE/g from the 1R-AM. The values of calycosin and formononetin, which are the known isoflavonoid constituents of AM, were 778.58±2.72 and 726.80±3.45 µg/g respectively, from the 2R-AM. Treatment of the HDF cells with R-AM (50 ~ 200 µg/mL) did not affect the cell viability. Furthermore, the R-AM extracts effectively protected against UVB-induced DNA damage.
Figures and Tables
Fig. 1
Chromatogram of AMs using HPLC analysis. (A) Scheme for AM samples. NR-AM, none roasted-Astragalus membranaceus; 1R-AM, 1st roasted-Astragalus membranaceus; 2R-AM, 2nd roasted-Astragalus membranaceus; 3R-AM, 3rd roasted-Astragalus membranaceus; B–F, Chromatogram of AMs using HPLC analysis; 1, daidzein-7-O-β-D-glucoside; 2, calycosin-7-O-β-D-glucoside; 3, formononetin-7-O-β-D-glucoside; 4, daidzein; 5, calycosin; 6, formononetin.
Fig. 2
Cell viability of AMs on HDF cells. HDF cells were treated R-AM samples of 50, 100 and 200 µg/mL for 24 h. Cell viability was examined using MTS assay. NR-AM, none roasted-Astragalus membranaceus; 1R-AM, 1st roasted-Astragalus membranaceus; 2R-AM, 2nd roasted-Astragalus membranaceus; 3R-AM, 3rd roasted-Astragalus membranaceus.
Fig. 3
Inhibitory effect of AMs during repair of UVB-induced DNA damage on HDF cells. HDF cells were cultured in 96-well plates and exposed to the indicated fluorescent of UVB. Single cells were embedded in agarose immediately after UV irradiation, electrophoresed and photographed using a fluorescent microscope to determine the “Tail moment” of each Tail length * % DNA in tail (representing the extent of DNA damage). NR-AM, none roasted-Astragalus membranaceus; 1R-AM, 1st roasted-Astragalus membranaceus; 2R-AM, 2nd roasted-Astragalus membranaceus; 3R-AM, 3rd roasted-Astragalus membranaceus; (A) Representative images of wild and UVB-induced HDF cells. (B) Tail DNA intensity was quantified by each 3 arrow points in the Fig. 1A. Results are significantly different among groups by Duncan's multiple range test (p < 0.05).
Table 1
Antioxidant activity of AMs
1) AM, Astragalus membranaceus; NR-AM, none roasted-Astragalus membranaceus; 1R-AM, 1st roasted-Astragalus membranaceus; 2R-AM, 2nd roasted-Astragalus membranaceus; 3R-AM, 3rd roasted-Astragalus membranaceus
2) L-ascorbic acid was used for positive control.
All values are represented as means± SD (n = 3). Results with different letters significantly different among groups are analyzed by Duncan's multiple range test (p < 0.05).
Table 2
Total phenolic contetns of AMs
1) AM, Astragalus membranaceus; NR-AM, none roasted-Astragalus membranaceus; 1R-AM, 1st roasted-Astragalus membranaceus; 2R-AM, 2nd roasted-Astragalus membranaceus; 3R-AM, 3rd roasted-Astragalus membranaceus
2) TPC, total phenol content; GAE, gallic acid equivalent All data are presented as the mean±SD of triplicate determinations. a,b,c: means in a row followed by different superscripts are significantly different (p < 0.05) by Duncan's multiple range test.
Table 3
The major isoflavonoid contents of AMs
1) AM, Astragalus membranaceus; NR-AM, none roasted-Astragalus membranaceus; 1R-AM, 1st roasted-Astragalus membranaceus; 2R-AM, 2nd roasted-Astragalus membranaceus; 3R-AM, 3rd roasted-Astragalus membranaceus
All data are presented as the mean± SD of triplicate determinations.
Value with different letters indicate significance between groups by Duncan's multiple range test (p < 0.05).
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