• Plant Biotechnology Reports
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• 제4권4호
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• pp.253-260
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• 2010

Plant secondary metabolites have always been a focus of study due to their important roles in human medicine and nutrition. We transferred the isoflavone synthase (IFS) gene into soybean [Glycine max (L.) Merr.] using the Agrobacterium-mediated transformation method in an attempt to produce transformed soybean plants which produced increased levels of the secondary metabolite, isoflavone. Although the trial to produce transgenic plant failed due to unestablished hygromycin selection, transformed callus cell lines were obtained. The induction rate and degree of callus were similar among the three cultivars tested, but light illumination positively influenced the frequency of callus formation, resulting in a callus induction rate of 74% for Kwangan, 67% for Sojin, and 73% for Duyou. Following seven to eight subcultures on selection media, the isoflavone content of the transformed callus lines were analyzed by high-performance liquid chromatography. The total amount of isoflavone in the transformed callus cell lines was three- to sixfold higher than that in control callus or seeds. Given the many positive effects of isoflavone on human health, it may be possible to adapt our transformed callus lines for industrialization through an alternative cell culture system to produce high concentrations of isoflavones.

• Natural Product Sciences
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• 제5권3호
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• pp.113-120
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• 1999

Nitric oxide (NO) is a free radical synthesized from L-arginine by nitric oxide synthase (NOS). It is believed that NO is an important mediator in numerous physiological and inflammatory responses. Particularly, a large amount of NO released from the inducible nitric oxide synthase (iNOS) is mostly associated with inflammatory processes. Overproduction of NO in these processes including sepsis and autoimmune diseases can have deleterious consequences and pathophysiologic relevance. Therefore, for the discovery of new inhibitory agents against iNOS activity, we have evaluated about 100 kinds of natural products after partition into three layers (n-hexane, ethyl acetate and aqueous) from 100% methanol extracts to study inhibitory effects on iNOS activity induced by lipopolysaccharide (LPS) in RAW264.7 cells culture system. As a positive control, curcumin, which is known as an anti-tumor promoter, anti-inflammatory agent as an iNOS inhibitor, was used and showed the dose-dependent inhibitory effect $(IC_{50},\;2.5\;{\mu}g/ml)$. Among tested fractions, the n-hexane fraction of Cimicifuga heracleifolia $(IC_{50}:\;9.65\;{\mu}g/ml)$, Forsythiae fructus $(IC_{50}:\;6.36\;{\mu}g/ml)$, Saposhnikovia divaricata $(IC_{50}:\;5.92\;{\mu}g/ml)$, and the ethyl acetate fraction of Chrysanthemum sibiricum $(IC_{50}:\;2.56\;{\mu}g/ml)$, Gastrodia elata $(IC_{50}:\;3.46\;{\mu}g/ml)$, and the aqueous fraction of Dianthus chinensis $(IC_{50}:\;6.73\;{\mu}g/ml)$, Euonymus alatus $(IC_{50}:\;6.78\;{\mu}g/ml)$, Mechania urticifoloria $(IC_{50}:\;8.01\;{\mu}g/ml)$ showed strong inhibitory activity against LPS-stimulated iNOS. Especially, the ethyl acetate fraction of Chrysanthemum sibiricum $(IC_{50}:\;2.56\;{\mu}g/ml)$, which exhibited the strongest inhibition against iNOS, was fractionated with silica-gel column chromatography. These subfractions exhibited dose-dependent inhibition against iNOS activity in the range of $2.59-5.6\;{\mu}g/ml$ except for fraction No. 3, 4, 5, 6, 8, 9, and 16. Our study shows that Chrysanthemum sibiricum has the strongest inhibitory effect against iNOS activity and has similar effect to curcumin. Therefore, further studies for the identification of active principles from Chrysanthemum sibiricum and investigation for the mechanism of the inhibition of iNOS by active principles will be performed.

• Natural Product Sciences
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• 제11권2호
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• pp.67-74
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• 2005

The ethanolic extracts of the leaves and bulbs of Allium victorialis var. platyphyllum (Liliaceae) collected from Daegwallyoung (D) and Ullung Island (U) in Korea were obtained using three different extracting methods. The first extracts, DL-1 DB-1, UL-1 and UB-1, were obtained from leaves (L) and bulbs (B) dried at $90^{\circ}C$, respectively, and the second extracts, DL-2, DB-2, UL-2 and UB-2, were obtained by extracting the leaves and bulbs of fresh plant parts. The third extracts DL-3, DB-3, UL-3 and UB-3 were obtained by incubating leaves and bulbs at $36^{\circ}C$. The six extracts obtained from A. victorialis var. platyphyllum at Daegwanllyoung (cultivated site) were orally administered to examine for a possible antihepatotoxic effect in $CCl_4-induced$ rats. DL-1 exhibited the most pronounced effect. The extracts inhibited serum ALT, AST, SDH, ${\gamma}-GT$, ALP and LDH activities elevated by $CCl_4$ injection and attenuated decreased glutathione S-transferase, glutatione reductase and ${\gamma}-glutamylcysteine$ synthetase activities and a decreased hepatic glutathione. However, the extracts obtained from Ullung Is. (native site) were less active than the extracts from Daegwallyoung, suggesting that A. victorialis var. platyphyllum from the cultivated site is more useful for functional food than of native site. These results also suggest that the antihepatotoxic effect is due to a higher content of hepatic glutathione. Gas chromatography of the twelve extracts showed significantly different sulfides, disulfides or trisulfides contents belonging to volatile sulfur substances (VSS). Nine components were identified on the basis of their mass spectra, namely, dimethyl disulfide, dimethyl trisulfide, diallyl disulfide, dipropyl disulfide, allyl methyl sulfide, allyl methyl trisulfide, 2-vinyl-4H-1,3-dithiin, 3,4-dihydro-3-vinyl-1,2-dithiin, and allithiamine. Extract DL-1 had the highest VSS content. Dried plant materials contained larger amounts of the VSSs than other extracts, and the leaves contained larger amount than the bulbs. These results suggest that heat treatment increases the antiheaptotoxic ability of A. victorialis var. platyphyllum by increasing the proportion of VSSs.

• Natural Product Sciences
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• 제11권2호
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• pp.103-108
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• 2005

Bioflavone quercetin is thought to have an important role to inhibit bone loss by affecting osteoclastogenesis and regulating a number of systemic and local factors such as hormones and cytokines. In this study, we examined how quercetin acts on cytokine production and mineralization of osteoblast in the presence of tumor necrosis factor-alpha $(TNF-{\alpha})$ which has been known to play a pivotal role in bone metabolic diseases. Quercetin inhibited $TNF-{\alpha}-induced$ secretion of $IFN-{\gamma}$ and IL-6 in differentiated MC3T3-E1 cells. As indicated by the markers that are characteristics of the osteoblast phenotype, such as alkaline phosphatase (ALP) activity and calcium deposition, quercetin treatment slightly prevented the $TNF-{\alpha}-induced$ dramatic inhibition of differentiation and mineralization of MC3T3-E1 cells. Further, quercetin inhibited the production of nitric oxide induced by $TNF-{\alpha}$ in the cells. Collectively, our findings indicate that quercetin inhibites $TNF-{\alpha}-induced$ secretion of inflammatory cytokines in differentiated MC3T3-E1 cells without any cytotoxic effects.

• Natural Product Sciences
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• 제21권3호
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• pp.150-154
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• 2015

Phytochemical investigation of Kandelia candel resulted in the isolation of six triterpenes (1 - 5) and two glyceryl glycosides (6 and 7) and their structures were determined by comparing the spectroscopic data with those of reported values. In present study, we described the inhibitory effects of fractions and isolated compounds from K. candel on pro-inflammatory cytokines (IL-12 p40, IL-6, and TNF-α) production in lipopolysaccharide (LPS) stimulated bone marrow-derived dendritic cells (BMDCs). Results indicated that compounds 3, 6, and 7 showed potent inhibition on IL-6 production (IC₅₀ values at less than 0.5 μM, respectively). Meanwhile, compounds 6 and 7 exhibited strong inhibitory effects on the production of TNF-α (IC₅₀ values of 1.7 ± 0.1 and 5.5 ± 0.2 μM). Compounds 1 and 3 were also showed the inhibitory effects on IL-12 p40 production (IC₅₀ values of 8.9 ± 0.4 and 3.3 ± 0.1 μM, respectively).

• Natural Product Sciences
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• 제20권3호
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• pp.137-145
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• 2014

The biological activities of licorice F1 (Glycyrrhiza glabra ${\times}$ G. uralensis) lines (G) were investigated, revealing strong radical scavenging activity targeting 1,1-diphenyl-2-picrylhydrazyl (DPPH) and hydroxyl (${\cdot}OH$) radicals. At a concentration of $100{\mu}g/mL$, most of the licorice F1 lines scavenged DPPH and ${\cdot}OH$ by more than 80%. Gs-1, -2, and -6 can be considered good scavengers of DPPH radical and G-7 have higher antioxidant activity against ${\cdot}OH$ radical. In addition, licorice F1 lines exerted effective anti-microbial activities against Escherichia coli (Gs-12, -17, and -18) and Staphylococcus aureus (Gs-3, -4, -5, -21, and -26). Moreover, Gs-2, - 20, -31, and -32 effectively inhibited the growth of Helicobacter pylori. Among licorice F1 lines, Gs-25 exhibited high anti-inflammatory effects on nitric oxide produced by lipopolysaccharide- and interferon-${\gamma}$-activated RAW 264.7 cells. Furthermore, Gs-1, -12, and -20 inhibited the growth of AGS human gastric adenocarcinoma cells by more than 60% at a concentration of $100{\mu}g/mL$ and Gs-5, -11, -19, and -32 showed inhibitory effects against rat lens aldose reductase ($IC_{50}$ values, 1.69, 6.07, 6.12, and $4.54{\mu}g/mL$, respectively). The total content of glycyrrhizin (1), glycyrrhetinic acid (2), glabridin (3), and isoliquiritigenin (4) in licorice F1 lines was high in Gs-11, -15, and -30. The present study therefore indicated that Gs-2, -26, -31, and -32 of licorice F1 possessing strong anti-oxidative, anti-microbial, anti-inflammatory, anti-cancer, and aldose reductase inhibitory effects may be used as a possible source material for natural health supplements in the future.

• Natural Product Sciences
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• 제19권1호
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• pp.36-48
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• 2013

The aim of the present study was to investigate the effects of several fractions obtained from methylene chloride ($CH_2Cl_2$) extract of self-fermented pine needle (SFPNE) on the acetylcholine (ACh)-evoked CA release from the isolated perfused model of the rat adrenal medulla and to establish the mechanism of the most active fraction (Fr.)-induced inhibitory action on the CA release. We obtained 6 fractions from $CH_2Cl_2$ extract of self-fermented pine needle. For the ACh (5.32 mM)-evoked CA release, the following rank order of inhibitory potency was obtained: Fr.4-5 > Fr.8-11 ${\gg}$ Fr.3 > Fr.6 = Fr.7 > Fr.1-2. Fr. 4 - 5 (60 ${\mu}g/mL$) perfused into an adrenal vein for 90 min produced relatively time-dependent inhibition of the CA secretory responses to ACh (5.32 mM), DMPP (100 ${\mu}M$), McN-A-343 (100 ${\mu}M$) and high $K^+$ (56 mM). Fr. 4 - 5 itself did not affect basal CA secretion. Also, in the presence of Fr. 4 - 5 (60 ${\mu}g/mL$), the CA secretory responses to angiotensin II (AngII, 0.1 ${\mu}M$), veratridine (50 ${\mu}M$), Bay-K-8644 (10 ${\mu}M$), and cyclopiazonic acid (10 ${\mu}M$) were significantly reduced, respectively. In the simultaneous presence of Fr. 4 - 5 (60 ${\mu}g/mL$) and L-NAME (30 ${\mu}M$), the inhibitory responses of Fr. 4 - 5 on the CA secretion evoked by ACh, DMPP, high $K^+$, AngII, Bay-K-8644 and veratridine were considerably recovered to the extent of the corresponding control secretion compared with that of Fr. 4 - 5-treatment alone. The level of NO released from adrenal medulla after the treatment of Fr. 4 - 5 (60 ${\mu}g/mL$) was greatly elevated compared with the basal level. Taken together, these results demonstrate that Fr. 4 - 5 inhibits the CA secretion from the isolated perfused rat adrenal medulla evoked by stimulation of cholinergic receptors as well as by direct membrane-depolarization. It seems that this inhibitory effect of Fr. 4 - 5 is mediated by blocking the influx of $Ca^{2+}$ and $Na^+$ into the adrenomedullary chromaffin cells as well as by inhibition of $Ca^{2+}$ release from the cytoplasmic calcium store, which is evoked at least partly through the increased NO production due to the activation of NO synthase. Based on these results, it is also thought that Fr. 4 - 5 isolated from $CH_2Cl_2$ extract of pine needle may contain beneficial antihypertensive components to prevent or treat hypertension.

• Natural Product Sciences
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• 제23권3호
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• pp.192-200
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• 2017

Skin aging is a complex biological process due to intrinsic and extrinsic factors. Free radical oxidative is one of extrinsic factors that induce activation of collagenase, elastase and hyaluronidase. Natural product from plants has been used as antioxidant and antiaging. This study aimed to evaluate antioxidant and antiaging properties of Hibiscus sabdariffa extract (HSE) and its compounds including myricetin, ascorbic acid, and ${\beta}$ carotene. The phytochemical of H. sabdariffa was determined using modified Farnsworth method and presence of phenols, flavonoids and tannins were in moderate content, whereas triterpenoids and alkaloids were in low content. Total phenolic content performed using Folin-Ciocalteu method, was $23.85{\mu}gGAE/mg$. Quantitative analysis of myricetin, ${\beta}-carotene$, and ascorbic acid of HSE was performed with Ultra-High Performance Liquid Chromatography (UHPLC) that shows $78.23{\mu}g/mg$ myricetin, $0.034{\mu}g/mg$ ${\beta}-carotene$, whilst ascorbic acid was not detected. HSE has lower activity on DPPH ($IC_{50}=195.73{\mu}g/mL$) compared to ${\beta}-carotene$, the lowest in ABTS assay ($IC_{50}=74.58{\mu}g/mL$) and low activity in FRAP assay ($46.24{\mu}MFe(II)/{\mu}g\;$) compared to myricetin, ${\beta}-carotene$. Antiaging was measured through inhibitory activity of collagenase, elastase, and hyaluronidase. HSE had weakest collagenase inhibitory activity ($IC_{50}=750.33{\mu}g/mL$), elastase inhibitory activity ($103.83{\mu}g/mL$), hyaluronidase inhibitory activity ($IC_{50}=619.43{\mu}g/mL$) compared to myricetin, ${\beta}-carotene$, and ascorbic acid. HSE contain higher myricetin compared to ${\beta}-carotene$. HSE has moderate antioxidants and lowest antiaging activities. Myricetin is the most active both antioxidant and antiaging activities.

• Natural Product Sciences
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• 제26권1호
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• pp.83-89
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• 2020

Osteoporosis is a worldwide disease leading to significant economic and societal burdens globally. Osteoporosis is caused by unbalanced bone remodeling between the rate of osteoclast bone resorption and osteoblast bone formation. Acer tegmentosum Maxim (AT) is a traditional herbal medicine containing multiple biological activities such as anti-oxidant and anti-inflammatory purposes. However, its role in osteoporosis has not been fully studied. Therefore, we investigated whether AT has a potent inhibitory effect on osteoporosis and its mechanism through a systemic evaluation in ovariectomized (OVX) mice. OVX mice were orally administrated with the AT at doses of 50, 100, and 200 mg/kg for 10 weeks. Histological images and histomorphometry analyses were performed by H&E and Toluidine blue satin, and the expression levels of receptor activator for nuclear factor-kB ligand (RANKL), nuclear factor of activated T cells cytoplasm 1 (NFATc1), c-Fos, and matrix metalloproteinase 9 (MMP9) related to the osteoclast differentiation were investigated using immunohistochemical analysis. Administration of AT prevented bone loss and the alternations of osteoporotic bone parameters at the distinct regions of the distal femur and spongiosa region in OVX mice. Further, administration of AT increased periosteal bone formation in a dose-dependent manner. Meanwhile, AT inhibited not only the expression of NFATc1 and c-Fos, which are two major regulators of osteoclastogenesis but also reduced bone resorbed encoding expression of MMP9 and RANKL. Our results indicated that administration of AT prevented bone loss and the alternations of osteoporotic bone parameters at the distinct regions of the distal femur and spongiosa region in OVX mice. Also AT has the bone protective effect through the suppression of osteoclast and promotion of osteoblast, suggesting that it could be a preventive and therapeutic candidate for anti-osteoporosis.

• 생명과학회지
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• 제16권6호
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• pp.949-954
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• 2006

콩은 전 세계 시장의 48%를 차지하는 중요한 유료작물이다. 콩 종자를 구성하는 양적인 부분과 질적인 부분의 개선은 콩 육종 목표의 중요한 부분이다. 단백질함량과 종실의 크기는 두부와 콩나물의 질을 평가하는 중요한 특성이다. 따라서 본 연구는 콩에서 종실의 크기와 단백질 및 oil 함량을 조절하는 양적형질유전자좌(QTLs)를 확인하기 위하여 실시하였다. 시험재료로는 큰올콩과 신팔달콩을 교배한 후 $F_2$유래 $F_10$세대의 RIL을 이용하였으며, 이를 바탕으로 종실의 무게와 oil 및 단백질 함량과 관련된 QTLs를 탐색하였다. 종실의 무게와 단백질 및 oil 함량의 협의의 유전력은 각각 0.8과 0.78 및 0.71을 나타내었다. 종실의 무게와 관련된 QTLs는 연관군 F, I와 K의 세 개의 독립적인 QTLs를 확인하였다. 단백질함량과 관련된 QTLs는 연관군 D1b, E, H, I와 L의 다섯 개의 독립적인 QTLs를 확인하였다. 그리고 oil 함량과 관련된 QTLs는 연관군 D1b, E, G, I, J와 N의 여섯 개의 독립적인 QTLs를 확인하였다. oil 및 단백질 함량과 관련된 QTLs는 연관군 D1b, E와 I에서 공통적으로 나타났다. 따라서 이들 주요 QTL은 콩 품종 육성과정에서 품질의 개선하기 위한 선발 마커로서 활용가치가 높은 것으로 판단된다.