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Studies about the bioactive component analysis and an oral glucose tolerance test of Add-Omit-Saenghyeoryunbu-eum(AO-SHU) for confirmation of diabetes therapy (가감생혈윤부음(加減生血潤膚飮)의 당뇨병 치료효과 확인을 위한 생리활성성분 분석과 경구포도당부하 연구)

  • In, Jeongdo;Im, Daisig;Kim, Won-Ill
    • Herbal Formula Science
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    • v.24 no.2
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    • pp.80-99
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    • 2016
  • Objectives : Instrumental chemical analysis was utilized to investigate the effect of Add-Omit-Saenghyeoryunbu-eum(AO-SHU) on diabetic treatment. One of the most exciting, yet also controversial, arguments is the safety and biological mechanisms of the natural medicine on human body. Therefore, the aim of this study is to provide a better understanding on bioactive chemical components, hazards of heavy metal contamination and biological mechanism of the diabetic medicine composed of 12 different natural herbs. Methods : To study bioactive compound and metallic component in the diabetic medicine in detail, LC-MS/MS (Liquid Chromatography-Mass/Mass), GC (Gas Chromatography) and ICP (Inductively Coupled Plasma) were utilized to characterize the extract of the diabetic medicine and the result was compared with 18 marker substances selected from literature survey. In addition, in vitro assay experiments including GPR 119 activity and human DGAT-1 inhibition, and OGTT (Oral Glucose Tolerance Test) were performed to verify the effectiveness of this medicine on diabetic treatment. Results : Out of 18 marker substances, 9 bioactive compounds were identified from LC-MS/MS analysis which include Citruline, Catalpol, Berberine, Ginsenoside Rb1, Ginsenoside Rg1, Oleanolic acid, β-Sitosterol, Mangiferin, and Schizandrin. ICP study on 245 residual pesticides revealed that 239 species were not detected but 6 species, Dimethomorph, Trifloxystrobin, Pyraclostrobin, Isoprocarb, Carbaryl and Flubendiamide, while the amounts are trace levels, below permitted concentrations. The biological activity was observed in vitro assay and Oral Glucose Tolerance Test(OGTT), which are consistent with a preliminary clinical test result, a drop in blood sugar level after taking this herbal medicine. Conclusions : Instrumental chemical analysis using LC-MS/MS, GC, and ICP was conducted successfully to identify bioactive compounds in AO-SHU for the treatment of diabetes, finding 9 bioactive compounds. Furthermore, in vitro assay experiments and OGTT show that AO-SHU has its biological activities, which imply that it can be a candidate for the future diabetes remedy.

Immunomodulatory and anti-metastatic activities of polysaccharide isolated from red cabbage (적양배추에서 분리한 다당의 면역 및 항전이 활성)

  • Lee, Sue Jung;Shin, Kwang-Soon
    • Korean Journal of Food Science and Technology
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    • v.51 no.3
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    • pp.263-271
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    • 2019
  • In this study, we examined the immunostimulating characteristics of a hot water extract (RCW) and crude polysaccharides (RCP) of red cabbage. RCW and RCP did not show any cytotoxicity in B16BL6 cells and macrophages. Although the sugar compositions of RCW and RCP were similar, the uronic acid content of RCP was higher than that of RCW RCP significantly increased the production of various cytokines and NO, whereas RCW did not affect the production of cytokines and NO. In an ex vivo assay of natural killer (NK) cell activity, intravenous (i.v.) administration of RCP significantly augmented NK cytotoxicity against Yac-1 tumor cells at 3 days after RCP treatment. In an experimental lung metastasis model using B16BL6 melanoma cells, i.v. administration of RCP at a dose of $1,000{\mu}g$ per mouse significantly inhibited 47.3% of lung metastasis. These results suggest that crude polysaccharide isolated from red cabbage is a promising food ingredient for the prevention of tumor metastasis.

Changes in Growth, Active Ingredients, and Rheological Properties of Greenhouse-cultivated Ginseng Sprout during its Growth Period (하우스에서 재배된 새싹인삼의 재배시기별 생육, 유효성분 및 물성의 변화)

  • Seong, Bong Jae;Kim, Sun Ick;Jee, Moo Geun;Lee, Hee Chul;Kwon, A Reum;Kim, Hyun Ho;Won, Jun Yeon;Lee, Ka Soon
    • Korean Journal of Medicinal Crop Science
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    • v.27 no.2
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    • pp.126-135
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    • 2019
  • Background: The ginseng ginsenosides, which have various physiological activities, are known to be more abundant in the leaves than in the roots, and the consumers' interest in ginseng sprout as a functional vegetable has been increasing. Methods and Results: The aim of this study was to investigate the effects of growth period on growth properties, active ingredients and rheology of ginseng sprouts cultivated in a non-heated greenhouse equipped with a shade net for 60 days, starting from the end of May to the middle of July. The chlorophyll content of the leaves decreased, but their length and width increased with increasing cultivation days. In particular, growth increased significantly until 40 days, but only slightly after 50 days. The stem length did not increase greatly from the 20 th to the 30 th day of cultivation, but increased significantly from the 30 th to the 40 th day, and then further increased gradually. The weight of the leaves, stems, and roots increased slightly, but not change significantly. After 40 days of cultivation, the total ginsenoside content increased by 1.07 times in the leaves and decreased by 0.80 times in the roots with increasing cultivation days. The leaf contents of ginsenosides $Rg_1$, Re, $Rb_1$, Rc, $F_3$ and $F_4$ increased with increasing cultivation days. The rheological properties of ginseng sprout showed the greatest influence on stem hardening with increasing cultivation days. Conclusions: Therefore, based on the growth characteristics, active ingredients and physical properties, 40 days after sowing was considered to be an appropriate harvesting time for ginseng sprouts.

A novel protocol for batch-separating gintonin-enriched, polysaccharide-enriched, and crude ginsenoside-containing fractions from Panax ginseng

  • Rami Lee;Han-Sung Cho;Ji-Hun Kim;Hee-Jung Cho;Sun-Hye Choi;Sung-Hee Hwang;Hyewon Rhim;Ik-Hyun Cho;Man-Hee Rhee;Do-Geun Kim;Hyoung-Chun Kim;Seung-Yeol Nah
    • Journal of Ginseng Research
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    • v.47 no.3
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    • pp.366-375
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    • 2023
  • Background: Ginseng contains three active components: ginsenosides, gintonin, and polysaccharides. After the separation of 1 of the 3 ingredient fractions, other fractions are usually discarded as waste. In this study, we developed a simple and effective method, called the ginpolin protocol, to separate gintonin-enriched fraction (GEF), ginseng polysaccharide fraction (GPF), and crude ginseng saponin fraction (cGSF). Methods: Dried ginseng (1 kg) was extracted using 70% ethanol (EtOH). The extract was water fractionated to obtain a water-insoluble precipitate (GEF). The upper layer after GEF separation was precipitated with 80% EtOH for GPF preparation, and the remaining upper layer was vacuum dried to obtain cGSF. Results: The yields of GEF, GPF, and cGSF were 14.8, 54.2, and 185.3 g, respectively, from 333 g EtOH extract. We quantified the active ingredients of 3 fractions: L-arginine, galacturonic acid, ginsenosides, glucuronic acid, lysophosphatidic acid (LPA), phosphatidic acid (PA), and polyphenols. The order of the LPA, PA, and polyphenol content was GEF > cGSF > GPF. The order of L-arginine and galacturonic acid was GPF >> GEF = cGSF. Interestingly, GEF contained a high amount of ginsenoside Rb1, whereas cGSF contained more ginsenoside Rg1. GEF and cGSF, but not GPF, induced intracellular [Ca2+]i transient with antiplatelet activity. The order of antioxidant activity was GPF > GEF = cGSF. Immunological activities (related to nitric oxide production, phagocytosis, and IL-6 and TNF-α release) were, in order, GPF > GEF = cGSF. The neuroprotective ability (against reactive oxygen species) order was GEF > cGSP > GPF. Conclusion: We developed a novel ginpolin protocol to isolate 3 fractions in batches and determined that each fraction has distinct biological effects.

Effects of Puffing Red Ginseng Powder on Antioxidant Enzyme Activities in Benzo(α)Pyrene-Treated Mice (팽화홍삼분말이 벤조피렌을 투여한 마우스의 항산화 효소 활성에 미치는 영향)

  • Kim, Hyun-Jeong;Lee, Ki-Dong;Lee, In-Seon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.7
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    • pp.847-852
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    • 2008
  • In order to determine the effects of puffing red ginseng (PRG) powder on the antioxidant enzyme activities of hepatotoxicity in benzo($\alpha$) pyrene[B($\alpha$)P]-treated mice, the mice were divided into 5 groups. The dried red ginseng were prepared by puffing conditions of moisture content 10% and puffing pressure $5\;kgf/cm^2$, and then powdered. PRG powder was injected i.p. once a day for 5 successive days, followed by the administration of B($\alpha$)P treatment on the fifth day. We also evaluated the relationship between lipid peroxidation and PRG powder on oxidative stress. The increased activities of superoxide dismutase, catalase, and glutathione peroxidase observed following B($\alpha$)P-treatment were reduced by the treatment of PRG powder. Whereas the glutathione content and glutathione S-transferase activity depleted by B $\alpha$)P were significantly increased, the B($\alpha$)P-associated elevation of cytochrome P-450 activities and lipid peroxide content were reduced as the result of PRG powder treatment. Especially, PRG powder had higher antioxidant activities than RG powder. These results suggest that puffing red ginseng powder can protect against B($\alpha$)P intoxicification through its antioxidant properties.

Metabolomes and transcriptomes revealed the saponin distribution in root tissues of Panax quinquefolius and Panax notoginseng

  • Wei, Guangfei;Yang, Feng;Wei, Fugang;Zhang, Lianjuan;Gao, Ying;Qian, Jun;Chen, Zhongjian;Jia, Zhengwei;Wang, Yong;Su, He;Dong, Linlin;Xu, Jiang;Chen, Shilin
    • Journal of Ginseng Research
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    • v.44 no.6
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    • pp.757-769
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    • 2020
  • Background: Panax quinquefolius and Panax notoginseng are widely used and well known for their pharmacological effects. As main pharmacological components, saponins have different distribution patterns in the root tissues of Panax plants. Methods: In this study, the representative ginsenosides were detected and quantified by desorption electrospray ionization mass spectrometry and high-performance liquid chromatography analysis to demonstrate saponin distribution in the root tissues of P. quinquefolius and P. notoginseng, and saponin metabolite profiles were analyzed by metabolomes to obtain the biomarkers of different root tissues. Finally, the transcriptome analysis was performed to demonstrate the molecular mechanisms of saponin distribution by gene profiles. Results: There was saponin distribution in the root tissues differed between P. quinquefolius and P. notoginseng. Eight-eight and 24 potential biomarkers were detected by metabolome analysis, and a total of 340 and 122 transcripts involved in saponin synthesis that were positively correlated with the saponin contents (R > 0.6, P < 0.05) in the root tissues of P. quinquefolius and P. notoginseng, respectively. Among them, GDPS1, CYP51, CYP64, and UGT11 were significantly correlated with the contents of Rg1, Re, Rc, Rb2, and Rd in P. quinquefolius. UGT255 was markedly related to the content of R1; CYP74, CYP89, CYP100, CYP103, CYP109, and UGT190 were markedly correlated with the Rd content in P. notoginseng.

Increase of Solubility of Ginseng Radix by Extrusion Cooking (압출성형 공정에 의한 인삼의 수용성 성분 증대)

  • Jee, Ho-Kyun;Cho, Young-Jin;Kim, Chong-Tai;Jang, Young-Sang;Kim, Chul-Jin
    • Korean Journal of Food Science and Technology
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    • v.38 no.3
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    • pp.361-368
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    • 2006
  • Extruded ginseng was manufactured using twin-screw extruder under 300 rpm screw speed, 21 kg/hr feed rate, $80-150^{\circ}C$ extrusion temperature, and by addition of water (12.1-30.6%). Extraction yield and contents of total carbohydrate and uronic acid in extruded ginseng at room temperature extraction (RT) increased with increasing extrusion temperature, whereas those of boiling temperature extracts (BT) were not affected by increasing extrusion temperature. Contents of nonstarch polysaccharide (NSP) in RT and BT extracts increased 340 and 142%, respectively, compared to that of raw ginseng. Main sugar compositions of NSP in RT and BT extracts were arabinose, galactose, and glucose. Extraction yields of total and crude saponins in extruded ginseng at optimize extrusion condition were higher than that of raw ginseng. In RT extracts, molecular weights of polysaccharides from raw were higher than that of extruded ginseng polymer, whereas in BT extracts molecular weights of polysaccharides from extruded ginseng were higher than those of raw ginseng polysaccharides.

Generation of a High-Growth Influenza Vaccine Strain in MDCK Cells for Vaccine Preparedness

  • Kim, Eun-Ha;Kwon, Hyeok-Il;Park, Su-Jin;Kim, Young-Il;Si, Young-Jae;Lee, In-Won;Kim, Se mi;Kim, Soo-In;Ahn, Dong-Ho;Choi, Young-Ki
    • Journal of Microbiology and Biotechnology
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    • v.28 no.6
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    • pp.997-1006
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    • 2018
  • As shown during the 2009 pandemic H1N1 (A(H1N1)pdm09) outbreak, egg-based influenza vaccine production technology is insufficient to meet global demands during an influenza pandemic. Therefore, there is a need to adapt cell culture-derived vaccine technology using suspended cell lines for more rapid and larger-scale vaccine production. In this study, we attempted to generate a high-growth influenza vaccine strain in MDCK cells using an A/Puerto/8/1934 (H1N1) vaccine seed strain. Following 48 serial passages with four rounds of virus plaque purification in MDCK cells, we were able to select several MDCK-adapted plaques that could grow over $10^8PFU/ml$. Genetic characterization revealed that these viruses mainly had amino acid substitutions in internal genes and exhibited higher polymerase activities. By using a series of Rg viruses, we demonstrated the essential residues of each gene and identified a set of high-growth strains in MDCK cells ($PB1_{D153N}$, $M1_{A137T}$, and $NS1_{N176S}$). In addition, we confirmed that in the context of the high-growth A/PR/8/34 backbone, A/California/7/2009 (H1N1), A/Perth/16/2009 (H3N2), and A/environment/Korea/deltaW150/2006 (H5N1) also showed significantly enhanced growth properties (more than $10^7PFU/ml$) in both attached- and suspended-MDCK cells compared with each representative virus and the original PR8 vaccine strain. Taken together, this study demonstrates the feasibility of a cell culture-derived approach to produce seed viruses for influenza vaccines that are cheap and can be grown promptly and vigorously as a substitute for egg-based vaccines. Thus, our results suggest that MDCK cell-based vaccine production is a feasible option for producing large-scale vaccines in case of pandemic outbreaks.

Effects of Korean Red Ginseng extract on tissue plasminogen activator and plasminogen activator inhibitor-1 expression in cultured rat primary astrocytes

  • Ko, Hyun Myung;Joo, So Hyun;Kim, Pitna;Park, Jin Hee;Kim, Hee Jin;Bahn, Geon Ho;Kim, Hahn Young;Lee, Jongmin;Han, Seol-Heui;Shin, Chan Young;Park, Seung Hwa
    • Journal of Ginseng Research
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    • v.37 no.4
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    • pp.401-412
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    • 2013
  • Korean Red Ginseng (KRG) is an oriental herbal preparation obtained from Panax ginseng Meyer (Araliaceae). To expand our understanding of the action of KRG on central nervous system (CNS) function, we examined the effects of KRG on tissue plasminogen activator (tPA)/plasminogen activator inhibitor-1 (PAI-1) expression in rat primary astrocytes. KRG extract was treated in cultured rat primary astrocytes and neuron in a concentration range of 0.1 to 1.0 mg/mL and the expression of functional tPA/PAI-1 was examined by casein zymography, Western blot and reverse transcription-polymerase chain reaction. KRG extracts increased PAI-1 expression in rat primary astrocytes in a concentration dependent manner (0.1 to 1.0 mg/mL) without affecting the expression of tPA itself. Treatment of 1.0 mg/mL KRG increased PAI-1 protein expression in rat primary astrocytes to $319.3{\pm}65.9%$ as compared with control. The increased PAI-1 expression mediated the overall decrease in tPA activity in rat primary astrocytes. Due to the lack of PAI-1 expression in neuron, KRG did not affect tPA activity in neuron. KRG treatment induced a concentration dependent activation of PI3K, p38, ERK1/2, and JNK in rat primary astrocytes and treatment of PI3K or MAPK inhibitors such as LY294002, U0126, SB203580, and SP600125 (10 ${\mu}M$ each), significantly inhibited 1.0 mg/mL KRG-induced expression of PAI-1 and down-regulation of tPA activity in rat primary astrocytes. Furthermore, compound K but not other ginsenosides such as Rb1 and Rg1 induced PAI-1 expression. KRG-induced up-regulation of PAI-1 in astrocytes may play important role in the regulation of overall tPA activity in brain, which might underlie some of the beneficial effects of KRG on CNS such as neuroprotection in ischemia and brain damaging condition as well as prevention or recovery from addiction.

Effects of Cultured Wild Ginseng Roots on the Alcoholic Fermentation (산삼배양근 첨가가 알콜 발효에 미치는 영향)

  • Jeong Heon-Sang;Kang Tae-Su;Woo Koan-Sik;Paek Kee-Yeoup;Yu Kee-Won;Yang Seung-Joon
    • Food Science and Preservation
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    • v.12 no.4
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    • pp.402-410
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    • 2005
  • In order to manufacture the alcoholic drinks using cultured wild ginseng roots(CWGR) of 5 and $10\%$ (w/v), sugar content of fermentation media was adjusted to 24-25 $^{\circ}$Brix with white sugar and glucose. And 3 kinds of yeast (S. cerevisiae(KCCM 50757), S. cerevisiae (KCCM 50583) and S. bayanus(ATCC 10601) were used and then the quality of alcoholic drinks was analyzed by physical, chemical and sensory evaluation. Alcohol content was highest value of $15.8\%$ in $10\%$ of CWGR, white sugar, and S. bayanus(ATCC 10601). Major alcohols were ethanol and 1-propanol. Number of yeast cells increased to 5 days fermentation and slightly decreased afterwards. The pH was decreased abruptly from 5.0 in initial fermentation to 3.1-4.1 in 5 days fermentation. Total sugar contents were decreased continuously with fermentation periods and showed 7.0-10.5 $^{\circ}$Brix in 20 days fermentation. Saponin patterns and contents were various and higher in wine treated with S. bayanus(ATCC 10601). From the sensory evaluation, the highest score of overall quality was observed in the alcoholic beverage of $10\%$(w/v) of CWGR, glucose, and S. cerevisiae(KCCM 50583).