• Title/Summary/Keyword: In vitro plants

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Inhibitory Effect of Scopoletin Isolated from Sorbus commixta on TNF-α-Induced Inflammation in Human Vascular Endothelial EA.hy926 Cells through NF-κB Signaling Pathway Suppression (마가목 수피에서 분리한 scopoletin의 EA.hy926 혈관내피세포에서 NF-κB 신호전달을 통한 TNF-α로 유도된 혈관염증 저해 효과)

  • Kang, Hye Ryung;Kim, Hyo Jung;Kim, Bomi;Kim, Sun-Gun;So, Jai-Hyun;Cho, Soo Jeong;Kwon, Hyun Sook
    • Journal of Life Science
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    • v.30 no.4
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    • pp.343-351
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    • 2020
  • Sorbus commixta Hedl. has traditionally been used as a remedy for cough, asthma, and other bronchial disorders. In this study, three major triterpenoids-lupeol, β-sitosterol, and ursolic acid and a coumarin, scopoletin, were isolated from a CHCl3-soluble fragment of the bark of S. commixta. Their structures were identified by spectroscopic analyses, including mass spectrometry (MS), 1D-, and 2D- nuclear magnetic resonance spectroscopy (NMR), as well as by comparing the data with data reported in the literature. Scopoletin was isolated from this plant for the first time. It is a nutraceutical compound contained in many plants that has been reported to exert diverse biological activities, including anti-inflammatory effects. This study examined the inhibitory effect of scopoletin on TNF-α-induced vascular endothelial inflammation. Unlike the marginal impact of other compounds against low-density lipoprotein (LDL) oxidation and vascular endothelial inflammation, scopoletin showed remarkable activity on LDL oxidation (IC50 = 10.2 μM) and exerted vascular anti-inflammatory effects in EA.hy926 human endothelial cells activated by TNF-α. It suppressed the expression of adhesion molecules, such as ICAM-1, VCAM-1, and E-selectin, and blocked the adhesion between THP-1 monocytes and EA. hy926 endothelial cells. It also inhibited TNF-α-induced NF-κB translocation from the cytosol to the nucleus. Moreover, IκBα phosphorylation, which was increased by TNF-α treatment, was reduced after treatment with scopoletin. Thus, scopoletin inhibited TNF-α-induced vascular inflammation in endothelial cells by suppressing the NF-κB signaling pathway. These results demonstrate that owing to its anti-inflammatory activity in the vascular endothelium, scopoletin has the potential to inhibit atherosclerosis development.

In vitro Culture and Acclimatization of Regenerated Plants of Liliem cernum $K_{OMAROV}$ (솔나리 기내배양 및 재분화 식물체의 토양순화)

  • Kim, H.K.;Lim, Jung-Dae;Hyun, Tae-Kyung;Lee, Hyeon-Yong;Lee, Jin-Ha;Yu, Chang-Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.9 no.4
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    • pp.310-317
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    • 2001
  • The regenerated-bulblets placed in liquid free media resulted in good formation of roots and bulblets. On 1/4 MS free medium, roots and bulblets were predominantly induced. The 1/4 MS liquid medium supplemented with plant growth regulators was the best suitable condition for elongation of leaves and roots. Somatic embryos were frequently developed from embryogenic callus in liquid media with 2,4-D 1mg/ l . On free liquid media, the viability of callus reduced. As the salt strength of MS media reduces, the viability of callus reduced significantly. However, Leaves were induced from several callus clumps. When leaves, roots and bulb-scale segments were placed on MS media containing NAA 1mg/ l or 2,4-D 1mg/ l and various sucrose concentration, the best result about the differentiation, growth of leaf and the differentiation of leaf was obtained on MS media added 1.5% sucrose and 2,4-D 1mg/ l, 3% sucrose and NAA 1mg/ l, and 1.5% sucrose and NAA 1mg/ l, respectively. Also the better result differentiation, growth of root and differentiation of bulb was obtained on MS media with 6% sucrose and NAA 1mg/ l. Spermidine promoted the growth of leaf and the differentiation of bulb. However, spermine promoted the differentiation of leaf, the differentiation and the growth of root in MS solid media. On the MS liquid media, both spermine and spermidine stimulated organogenesis from bulb-scale segments. Regenerated plantlets were acclimatizated and grown in greenhouse in vermiculite + perlite (1 : 1 by volume) well. The optimal soil condition of rooting for plantlets regenerated was in peat moss.

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Evaluation of in-vitro Antithrombosis Activity of Lees of Korean Traditional Wine (전통주 주박의 항혈전 활성 평가)

  • Kim, Mi-Sun;Lee, Ye-Seul;Kim, Jong Sik;Shin, Woo-Chang;Sohn, Ho-Yong
    • Journal of Life Science
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    • v.24 no.8
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    • pp.865-872
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    • 2014
  • In this study, ethanol and hot water extracts of lees from Korean traditional wine (J-B, J-S, J-Y, J-H, and J-W) were prepared, and their effects on blood coagulation, platelet aggregation, and hemolysis of human red blood cells (hRBCs) were investigated to develop functional food ingredients from lees. The pH and brix of the lees ranged from 3.90 to 4.29 and 5.0 to $27.0^{\circ}$, respectively, and there was a huge difference in the water and ethanol content among the lees. The nuruk and additives used affected the color and physicochemical properties of lees. The J-W takju made from only rice and traditional nuruk, which has $13^{\circ}$ brix and 1.8% of alcohol, has potential as functional food ingredient. With regard to the extraction yields of lees, higher yields were obtained from J-H, which contains different medicinal plants, in ethanol, followed by J-W, J-B, J-S, and J-Y. Higher extraction yields of lees were obtained from J-S in hot water, followed by J-B, J-W, J-H, and J-Y, respectively. The ethanol extract of J-H and the hot water extract of J-Y had the highest contents of total polyphenol and total flavonoids among the lees extracts. The 10 lees extracts did not show hemolysis activity against hRBCs up to 5 mg/ml. In an anticoagulation activity assay, the ethanol extracts of three yakju lees (J-B, J-S, and J-Y) and the hot water extract of J-W inhibited thrombin activity, whereas the hot water extract of J-B, J-S, and J-H inhibited blood coagulation factors. In an antiplatelet aggregation activity assay, only the J-W takju lees showed significant inhibition activity. Our results suggest that lees from traditional wine had high potential as a novel antithrombosis agent.

The effect of two Terpenoids, Ursolic acid and Oleanolic acid on epidermal permeability barrier and simultaneously on dermal functions

  • Lim Suk Won;Jung Sung Won;Ahn Sung Ku;Kim Bora;Ryoo Hee Chang;Lee Seung Hun
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.29 no.2 s.43
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    • pp.205-232
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    • 2003
  • Ursolic acid (UA) and Oleanolic acid (ONA), known as urson, micromerol and malol, are pentacyclic triterpenoid compounds which naturally occur in a large number of vegetarian foods, medicinal herbs, and plants. They may occur in their free acid form or as aglycones for triterpenoid saponins, which are comprised of a triterpenoid aglycone, linked to one or more sugar moieties. Therefore UA and ONA are similar in pharmacological activity. Lately scientific research, which led to the identification of UA and ONA, revealed that several pharmacological effects, such as antitumor, hepato-protective, anti-inflammatory, anticarcinogenic, antimicrobial, and anti-hyperlipidemic could be attributed to UA and ONA. Here, we introduced the effect of UA and ONA on acutely barrier disrupted and normal hairless mouse skin. To evaluate the effects of UA and ONA on epidermal permeability barrier recovery, both flanks of 8-12 week-old hairless mice were topically treated with either 0.01-0.1 mg/ml UA or 0.1-1 mg/ml ONA after tape stripping, and TEWL (Transepidermal water loss) was measured . The recovery rate increased in those UA or ONA treated groups (0.1 mg/ml UA and 0.5 mg/ml ONA) at 6 h more than $20\%$ compared to vehicle treated group (p<0.05). Here, we introduced the effects of UA and ONA on acute barrier disruption and normal epidermal permeability barrier function. For verifying the effects of UA and ONA on normal epidermal barrier, hydration and TEWL were measured for 1 and 3 weeks after UA and ONA applications (2mg/ml per day). We also investigated the features of epidermis and dermis using electron microscopy (EM) and light microscopy (LM). Both samples increased hydration compared to vehicle group from f week without TEWL alteration (p<0.005). EM examination using RuO4 and OsO4 fixation revealed that secretion and numbers of lamellar bodies and complete formation of lipid bilayers were most prominent $(ONA{\geq}UA>Vehicle)$. LM finding showed that thickness of stratum corneum (SC) was slightly increased and especially epidermal thickening and flattening was observed (UA>ONA>Veh). We also observed that UA and ONA stimulate epidermal keratinocyte differentiation via $PPAR\;\alpha$. Protein expression of involucrin, loricrin, and filaggrin increased at least 2 and 3 fold in HaCaT cells treated with either $ONA\;(10{\mu}M)$ or UA $(10{\mu}M)$ for 24h respectively. This result suggested that the UA and ONA can improve epidermal permeability barrier function and induce the epidermal keratinocyte differentiation via $PPAR\;{\alpha}$. Using Masson-trichrome and elastic fiber staining, we observed collagen thickening and elastic fiber elongation by UA and ONA treatments. In vitro results of collagen and elastin synthesis and elastase inhibitory activity measurements were also confirmed in vivo findings. These data suggested that the effects of UA and ONA related to not only epidermal permeability barrier functions but also dermal collagen and elastic fiber synthesis. Taken together, UA and ONA can be relevant candidates to improve epidermal and dermal functions and pertinent agents for cosmeseutical applications.

Anti-inflammatory effects of Ishige sinicola ethanol extract in LPS-induced RAW 264.7 cell and mouse model (LPS로 유도된 RAW 264.7 Cell과 마우스 모델에 대한 넓패(Ishige sinicola) 에탄올 추출물의 항염증 효과)

  • Kim, Ji-Hye;Kim, Min-Ji;Kim, Koth-Bong-Woo-Ri;Park, Sun-Hee;Cho, Kwang-Su;Kim, Go-Eun;XU, Xiaotong;Lee, Da-Hye;Park, Ga-Ryeong;Ahn, Dong-Hyun
    • Food Science and Preservation
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    • v.24 no.8
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    • pp.1149-1157
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    • 2017
  • Inflammation is the first response of the immune system to infection or irritation in our body. The use of medicinal plants has been widely applied as an alternative source for drug development. One of marine natural resources, the anti-inflammatory effect of Ishige sinicola ethanol extract (ISEE), was evaluated by using LPS-induced RAW 264.7 cell and mice model. As a result, the production of nitric oxide (NO) and pro-inflammatory cytokines (IL-6, IL-$1{\beta}$, TNF-${\alpha}$) were inhibited with increasing concentration of ISEE without any cytotoxicity. Furthermore, ISEE suppressed the expression of not only inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), nuclear factor-kappa B (NF-${\kappa}B$) p65, and mitogen-activated protein kinases (MAPKs), including extracellular signal-regulated kinase (ERK) 1/2, p38, and c-Jun N-terminal kinase (JNK) in a dose-dependent manner. In mice ear edema test, the formation of edema was reduced at the highest dosage of ISEE and the reduction of the number of infiltrated mast cells was observed in histological analysis. These results indicate that ISEE has a potent anti-inflammatory activity and can be used as a pharmaceutical material for many kinds of inflammatory disease.

Total Polyphenol Contents and Antioxidant Activities of Methanol Extracts from Vegetables produced in Ullung Island (울릉도산 산채류 추출물의 총 폴리페놀 함량 및 항산화 활성)

  • Lee, Syng-Ook;Lee, Hyo-Jung;Yu, Mi-Hee;Im, Hyo-Gwon;Lee, In-Seon
    • Korean Journal of Food Science and Technology
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    • v.37 no.2
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    • pp.233-240
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    • 2005
  • To discover new functional materials using edible plants, antioxidant activities of methanol extracts from various parts of seven wild vegetables were investigated in vitro. Total polyphenol contents, determined by Folin-Denis method, varied from 16.74 to $130.22{\mu}g/mg$. Radical-scavenging activities of methanol extracts were examined using ${\alpha},\;{\alpha}-diphenyl-{\beta}-pirrylhydrazyl$ (DPPH) radicals and 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) assay. Inhibition effects on peroxidation of linoleic acid determined by ferric thiocyanate (FTC) method and on oxidative degradation of 2-deoxy-D-ribose in Fenton-type reaction system were dose-dependent. Athyrium acutipinulum Kodama (leaf and rood), Achyranthes japonica (Miq.) Nakai (seed), and Solidago virga-aurea var. gigantea Nakai (root) showed relatively high antioxidant activities in various systems.

In Vitro Assay on Biological Characteristics of Different Extracts from Acorus calamus L. var angustatus (창포(Acorus calamus L. var angustatus) 추출물의 생리활성 검정)

  • Heo, Buk Gu;Park, Yong Seo;Yoo, Yong Kweon;Han, Tae Ho;Park, Yun Jum;Sin, Jang Sik;Cho, Ja Yong
    • FLOWER RESEARCH JOURNAL
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    • v.16 no.3
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    • pp.168-173
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    • 2008
  • This study was conducted to determine biological characteristics of Acorus calamus L. angustatus by using aqueous, ethanol and methanol extracts from the leaves and the rhizomes of Acorus calamus L. angustatus plants. The highest total phenol contents were found in the extracts from the leaves of Acorus calamus L. angustatus ($68.4mg{\cdot}L^{-1}$) followed by rhizome ($49.3mg{\cdot}L^{-1}$). At $1,000mg{\cdot}L^{-1}$ the free radical scavenging activity of 1, 1-Diphenyl-2-picrylhydrazyl showed the highest activity in the extracts of the leaves and the rhizomes by 86.3% and 86.1%, respectively. Total flavonoid contents at $2,000mg{\cdot}L^{-1}$ extracts showed the greatest amount in the aqueous leaf extracts ($61.1mg{\cdot}L^{-1}$), and the ethanol rhizome extracts ($15.4mg{\cdot}L^{-1}$). Nitrite radical scavenging activity at $2,000mg{\cdot}L^{-1}$ was highest in the methanol extracts from leaves and rhizomes by 75.7% and 77.9%, respectively. Mushroom tyrosinase inhibition activity of leaves and rhizome from the Acorus calamus L. angustatus was very low, showing less than 1.3% regardless of the extract solvent, plant part, and concentration. These results indicated that methanol extracts from leaves exhibited higher biological activities than other extracts from rhizomes.

Elimination of Grapevine fleck virus from infected grapevines 'Kyoho' through meristem-tip culture of dormant buds (휴면아 경정 배양법을 통한 포도 '거봉' 에서 Grapevine fleck virus의 제거)

  • Kim, Mi Young;Cho, Kang Hee;Chun, Jae An;Park, Seo Jun;Kim, Se Hee;Lee, Han Chan
    • Journal of Plant Biotechnology
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    • v.44 no.4
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    • pp.401-408
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    • 2017
  • Herein, we report the meristem-tip culture from dormant buds of grape 'Kyoho' single-infected with Grapevine fleck virus (GFkV), which is phloem-limited and transmitted by graft inoculation. We produced GFkV-free shoots without thermo- or chemotherapy using meristem-tip explants approximately 0.3 mm (73 explants) and 0.8 mm long (five explants) including shoot apical meristem, 2-5 leaf primordia, and 1-4 uncommitted primordia from dormant buds of the infected woody cuttings (stored at $4^{\circ}C$). Explants were cultured on Murashige and Skoog (MS) medium supplemented with 3% sucrose, 3.0 mg/L benzyladenine (BA) and 0.1 mg/L indole-3-butyric acid (IBA). After 16 weeks of culture, shoot (10-mm long) regeneration frequency achieved from 0.3-mm explants was 4.1% and that obtained from 0.8-mm explants was 40.0%. Virus-free efficiency (expressed as the percentage of RT-PCR negative shoots regenerated) from 0.3- and 0.8-mm explants was 100% and 50%, respectively. Following in vitro multiplication, RT-PCR assays revealed identical results to assays of the first regenerated shoots. Our new methodological approach could be applied for eliminating other viruses in grapevines, as well as for producing virus-free plants in many other deciduous tree species, including fruit trees.

Metabolic comparison between standard medicinal parts and their adventitious roots of Cynanchum wilfordii (Maxim.) Hemsl. using FT-IR spectroscopy after IBA and elicitor treatment (IBA 및 elicitor 처리에 따른 백수오 기내 생산 부정근 및 표준품의 FT-IR 스펙트럼 기반 대사체 비교 분석)

  • Ahn, Myung Suk;So, Eun Jin;Jie, Eun Yee;Choi, So Yeon;Park, Sang Un;Moon, Byeong Cheol;Kang, Young Min;Min, Sung Ran;Kim, Suk Weon
    • Journal of Plant Biotechnology
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    • v.45 no.3
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    • pp.250-256
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    • 2018
  • To determine whether metabolite fingerprinting for whole cell extracts based on Fourier transform infrared spectroscopy (FT-IR) can be used to discriminate and compare metabolic equivalence, standard medicinal parts of Cynanchum wilfordii (Maxim.) Hemsl. and their adventitious roots were subjected to FT-IR. The principal component analysis (PCA) and partial least square discriminant analysis (PLS-DA) from FT-IR spectral data showed that whole metabolic pattern from the adventitious root of Cynanchum wilfordii was highly similar to its standard medicinal parts. These results clearly showed that mass proliferation of adventitious roots could be applied for the novel supply of standard medicinal parts of medicinal plants. Furthermore, FT-IR spectroscopy combined with multivariate analysis established in this study could be applied as an alternative tool for discriminating of whole metabolic equivalence from standard medicinal parts. Thus, it is proposed that these metabolic discrimination systems from the adventitious root of Cynanchum wilfordii could be applied for metabolic standardization of in vitro grown Cynanchum wilfordii.

Comparison of the Forage Quality and Productivity According to Varieties and Plant Parts of Imported Silage Corn (Zea mays, L) (도입 사일리지용 옥수수의 품종과 식물체 부위에 대한 사료가치와 생산성 비교)

  • Kim, Jong Geun;Li, Yan Feng;Wei, Sheng Nan;Jeong, Eun Chan;Kim, Hak Jin
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.40 no.2
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    • pp.98-105
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    • 2020
  • This experiment was conducted to a comparison of the productivity according to variety and forage quality by plant parts of imported silage corn (Zea mays, L) in Pyeongchang. The corns evaluated in this experiment were 8 varieties (P1184, P1151, P1194, P1543, P1345, P1429, P1443, and P2105) introduced from the United States, Pioneer Hybrid Co. The harvested corn was divided into 5 plant parts (leaf, stem, cob, husk, and grain), and the ratio of each part was calculated using dry weight and the feed value was analyzed. The emergence rate of corn was generally good except for the P1151 and P2105 varieties. The average tasseling date was July 24th and the silking date was July 27th, but the P2105 variety was late to July 28th and August 1st, and the remaining varieties were similar. P1345 was the highest (289 and 123 cm), and P1151 varieties were the lowest (267 and 101 cm) in the plant and ear height. Disease resistance was low in P1184, P1443 and P1429, and P1197 and P1345 were high. In the case of stover, the dry matter (DM) content was the lowest at 19.6% in the P1151 and the highest at 24.9% in the P1429. DM content of ear was the highest in the P2105 (55.5%), and P1184 (54.2%) and P1345 (54.3%) were also significantly higher (p<0.05). The DM yield of stover of P2105, P1429 and P1194 varieties was significantly higher (p<0.05), and ear yield of P2105, P1345 and P1443 was higher. The proportions of each part of plants (leaf, stem, cob, husk, and grain) divided by 5 was high, with 50-60% of the ear(grain+cob) ratio. The ratio of husk and cob was roughly similar, and the leaf and stem part showed a ratio of about 20%. The crude protein (CP) content was highest in leaf, followed by grain. The CP content of the stem was the lowest, and the husk was not significantly different among the varieties (p>0.05). The acid detergent fiber (ADF) content was similar to the rest parts except grain, but the leaf part tended to be lower, and other parts except the stem and leaf showed no significant difference between varieties (p>0.05). There was no significant difference in NDF (neutral detergent fiber) content in husk, but there was a difference between varieties in other parts (p<0.05). In addition, there was a special difference by plant parts for each variety, P2015 on the stem, P1197 on the leaf, P1151 on the cob, P1197 on the husk, and P1197 on the grains with high NDF content. IVDMD (in vitro dry matter digestibility) was not significantly different between stems and grains, but there was a difference between varieties in cobs and husks. According to the results, DM yield of P2105 variety was the best in the experiment, and the ratio of grain was excellent in P1543 and P1345. In addition, it was found that the feed value was higher in the leaves and grains, and the leaf and stem had higher feed values than husk or cob.