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Ginsenoside F4 inhibits platelet aggregation and thrombus formation by dephosphorylation of IP3RI and VASP

  • Shin, Jung-Hae;Kwon, Hyuk-Woo;Lee, Dong-Ha
    • Journal of Applied Biological Chemistry
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    • v.62 no.1
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    • pp.93-100
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    • 2019
  • The root of Panax ginseng is used in ethnomedicine throughout eastern Asia and various recent studies have proved that Panax ginseng has inhibitory effects on cardiovascular disease. Each factor causing cardiovascular disease is known to have a very complex process which is achieved by a diverse number of mechanisms. Among these factors, platelets are the most important because they directly participate in thrombogenesis. Therefore, inhibiting the activity of platelets is an essential element for prevention of cardiovascular diseases. Our previous study showed the antiplatelet effects of Korean red ginseng extract and two of its components, ginsenoside Rg3 and ginsenoside Ro. However, the inhibitory mechanism of other ginsenosides remains unclear. Therefore, we investigated the inhibitory mechanism of ginsenoside F4 (G-F4) from Korean red ginseng on the regulation of signaling molecules involved in human platelet aggregation. With the use of G-F4, collagen-induced human platelet aggregation was inhibited in a dose-dependent manner, and it suppressed collagen-induced elevation of $[Ca^{2+}]_i$ mobilization through elevated phosphorylation of inositol 1, 4, 5-triphosphate receptor I ($Ser^{1756}$). In addition, G-F4 inhibited fibrinogen binding to ${\alpha}IIb/{\beta}_3$ during collagen-induced human platelet aggregation. Thus, in the present study, G-F4 showed an inhibitory effect on human platelet activation, suggesting its potential use as a new natural medicine for preventing platelet-mediated cardiovascular diseases.

Antioxidant Activities of Ethanol Extracts from Black Ginseng Prepared by Steaming-Drying Cycles (흑삼 제조과정 중 증포 횟수에 따른 에탄올 추출물의 항산화 활성)

  • Kim, Hyo-Jin;Lee, Ji-Yeon;You, Bo-Ram;Kim, Hye-Ran;Choi, Jae-Eul;Nam, Ki-Yeul;Moon, Byung-Doo;Kim, Mee-Ree
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.2
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    • pp.156-162
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    • 2011
  • The objective of this study is to evaluate antioxidant activities of black ginseng prepared by nine repeated steaming-drying cycles. Ethanol extracts from each cycle of ginseng showed 33.5~41.0% of yields, 36.2~44.5% of moisture content and $64\sim66^{\circ}Brix$ of soluble solids. As the number of steaming-drying cycles increased, pH decreased, while the absorbance at 420 nm increased remarkably after the 4th cycle. Although the amounts of Rg1 and Rb1 contents quite decreased, the total phenol content of black ginseng (the final cycle of ginseng) was increased to 126%, compared with that of white ginseng. Antioxidant activities, determined by ferric-reducing antioxidant potential (FRAP), 2,2'-azinobis(3 ethybenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activity, 1,1-diphenyl-2-picrydrazyl (DPPH) and hydroxyl radical scavenging activities, increased remarkably as the number of steaming-drying cycles increased. Especially, FRAP value increased 155.6%. Also, $IC_{50}$ values for DPPH and hydroxyl radical scavenging activities of the final 9th-cycling product, decreased 4.5 folds and 9.7 folds, respectively, compared with those of white ginseng. Based on these results, it was suggested that antioxidant activities of black ginseng improve according to the increasing number of steaming-drying cycles, which was derived from increase of total phenol content.

Effect of $CO_2$ Enrichment on the Differentiation of Multi-shoots and Saponin contents in Tissue culture of Korean ginseng (Panax ginseng C. A. Meyer) (인삼(人蔘) 조직배양(組織培養)에서 $CO_2$처리(處理)가 multi-shoot 분화(分化) 및 사포닌 함량(含量)에 미치는 영향(影響))

  • Chung, Chan-Moon;Bae, Kil-Kwan
    • Korean Journal of Medicinal Crop Science
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    • v.7 no.4
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    • pp.296-302
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    • 1999
  • This experiment was conducted to study the effect of $CO_2$(0, 2, 500, 5, 000, 10, 000ppm) enrichment by enabling ventilation on micropropagation of multi-shoot and on the saponin contents in vitro in Korean ginseng (Panax ginseng C. A. Meyer). Embryo was cultured in Murashige and Skoog medium added 3mg/ l of Indolbutyric acid, Benzyladenin and Gibberellic acid $(GA_3)$, respectively. $CO_2$, enrichment had little effects on the number of adventitious buds and shoots originated from adventitious buds. The ratio of differentiated shoots to adventitious buds were about 50% in $CO_2$, enrichment treatment. The shoots originated from adventitious bud showed more rapid growth and had larger leaf area than the shoots originated from the leaf primordia did. The number of shoot primordia was the highest in 2, 500ppm of $CO_2$ enrichment treatment. On the contrary, 10,000ppm of $CO_2$, enrichment made smaller the number of shoot primordia and ratio of shoots to shoot primordia. The range of shoots differentiated was from shoot primordia were 15. 4 to 23. 9. The rate of dry weight of cultured shoots showed lowest (7. 5%) in control and highest (8. 59%) in 2, 500ppm of $CO_2$, enrichment. Rate of in vitro flower in control was 7.6% and that in 2500ppm of $CO_2$ was about twice (15.7-16.3%) as much as in control. Flower number per a embryo cultured was about 1.2-1.3. In the multi-shoots with callus enriched by 2, 500ppm of $CO_2$, the contents of crude saponin and ginsenosides in multi-shoots alone were higher than in multi-shoots with callus. The characteristics of ginsenosides in multi-shoots were especially the higher content of ginsenoside Rd, Re, and $Rg_1$.

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New Methods for Separation of Crude Ginseng Saponins (인삼 조사포닌의 새로운 분리 방법)

  • Shin, Ji-Young;Choi, Eon-Ho;Wee, Jae-Joon
    • Korean Journal of Food Science and Technology
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    • v.33 no.2
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    • pp.166-172
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    • 2001
  • In order to increase ginsenoside content, to reduce chemical change, to shorten extracting procedure, new methods of extraction and fractionation of crude ginseng saponin were established and compared for their chemical composition. Those are hot MeOH extraction/n-BuOH fractionation (BuOH method) and hot MeOH extraction/Diaion HP-20 adsorption/MeOH elution (HP-20 method), which are already known methods, and additional three new methods: hot MeOH extraction/cation AG 50W $adsorption/H_2O$ elution/n-BuOH extraction (AG 50W method), cool MeOH extraction/Diaion HP-20 adsorption/MeOH elution (cool extraction method) and direct extraction with EtOAc/n-BuOH (direct extraction method). AG 50W method provided a crude saponin showing the highest content of ginsenosides of 61.5% and the lowest contents of protein and free amino acids of 0.93% and 0.19%, respectively. The protein content was the highest as 14.18% in the crude saponin by HP-20 method, while free sugar content was the highest as 13.5% by BuOH method, indicating that these are factors that lower the rate of ginsenoside in crude saponins by those methods. On the other hand, it was revealed that AG 50W method produced large amount of prosapogenins during the pass through the cation exchange resin (AG 50W) column being strongly acidic. Crude saponin from direct extraction method showed relatively higher composition of ginsenoside $Rg_1$ and Re. The results suggest that contents and composition of ginsenosides and other chemical components in crude ginseng saponin greatly depend on the condition of the extraction and fractionation.

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Preparation and Quality Characteristics of the Fermentation product of Ginseng by Lactic Acid Bacteria (FGL) (유산균을 이용한 발효인삼 제조 및 품질 특성)

  • Park, Soo-Jin;Kim, Dong-Hyun;Paek, Nam-Soo;Kim, Sung-Soo
    • Journal of Ginseng Research
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    • v.30 no.2
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    • pp.88-94
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    • 2006
  • Ginseng as a raw material for production of probiotic ginseng product by lactic acid bacteria (LAB) was evaluated in this study. Either white ginseng (WG) or red ginseng (RG) (1% or 5%, w/v) were directly inoculated with a 24 hold seed culture of twenty seven substrains of four different LAB ($1.0{\times}10^6CFU/ml$); Lactobacillus spp., Streptococcus/Enterococcus spp., Leuconostoc/Lactococcus spp. and Bifidobacterium spp., and incubated at $37^{\circ}C$ for 24 or 48 h. Among 27 kinds of LAB, seven substrains of Lactobacillus (MG208, MG311, MG315, MG501, MG501C, MG505, MG590) and one Bifidobacterium (MG723) were selected based on their dose dependent stimulation of the growth of LAB in the presence of ginseng and changes in pH, acidity and viable cell counts during fermentation were examined. Lactobacillus MG208 specifically was found to show the best growth on 5% RG and reached nearly $14.0{\times}10^8CFU/ml$ after 48 h of fermentation and produced the titratable acidity as $0.84{\pm}0.02%$, whereas the pH was significantly lowered from $6.80{\pm}0.01\;to\;3.42{\pm}0.02$. These results indicated that ginseng can be an appropriate material to prepare the fermentation product by several strains of LAB. Therefore we should further check whether probiotic ginseng product may have synergistic health benefits of both probiotics and ginseng to serve for vegetarians and lactose-allergic consumers.

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).

The improvement of ginsenoside accumulation in Panax ginseng as a result of γ-irradiation

  • Kim, Dong Sub;Song, Mira;Kim, Sun-Hee;Jang, Duk-Soo;Kim, Jin-Baek;Ha, Bo-Keun;Kim, Sang Hoon;Lee, Kyung Jun;Kang, Si-Yong;Jeong, Il Yun
    • Journal of Ginseng Research
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    • v.37 no.3
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    • pp.332-340
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    • 2013
  • In this study, gamma rays were used to irradiate embryogenic calli induced from cotyledon explants of Panax ginseng Meyer. After the embryogenic calli were irradiated, they were transferred to adventitious roots using an induction medium; next, mutated adventitious root (MAR) lines with a high frequency of adventitious root formations were selected. Two MAR lines (MAR 5-2 and MAR 5-9) from the calli treated with 50 Gy of gamma rays were cultured on an $NH_4NO_3$-free Murashige and Skoog medium with indole-3-butyric acid 3 mg/L. The expression of genes related to ginsenoside biosynthesis was analyzed using reverse transcription polymerase chain reaction with RNA prepared from native ginseng (NG), non-irradiated adventitious root (NAR) and 2 MAR lines. The expression of the squalene epoxidase and dammarenediol synthase genes was increased in the MAR 5-2 line, whereas the phytosterol synthase was increased in the MAR 5-9 line. The content and pattern of major ginsenosides (Rb1, Rb2, Rc, Rd, Re, Rf, and Rg1) were analyzed in the NG, NAR, and 2 MAR lines (MAR 5-2 and MAR 5-9) using TLC and HPLC. In the TLC analysis, the ginsenoside patterns in the NG, NAR, and 2 MAR lines were similar; in contrast, the MAR 5-9 line showed strong bands of primary ginsenosides. In the HPLC analysis, compared with the NG, one new type of ginsenoside was observed in the NAR and 2 MAR lines, and another new type of ginsenoside was observed in the 2 MAR lines irradiated with gamma rays. The ginsenoside content of the MAR 5-9 line was significantly greater in comparison to the NG.

The Effect of Dried Roots of Rehmannia glutinosa Extract on Osteoblast in Rat Fetus Calvarial Cells (건지황(乾地黃) 추출물이 Rat fetus 두개골로부터 분리한 조골세포에 미치는 영향)

  • Im, Kyu-Jung;Choi, Kyung-Hee;Jung, Eun-Hye;Yoo, Dong-Youl
    • The Journal of Korean Obstetrics and Gynecology
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    • v.26 no.3
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    • pp.33-43
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    • 2013
  • Objectives: Osteoporosis is characterized by bone loss and morbidity with osteoporotic fracture. In this study, the author aimed to evaluate the effect of dried roots of Rehmannia glutinosa extract (RGE) on osteoblast proliferation in murine calvarial cells. Methods: The osteoblast separated from murine calvariae was cultivated for 6 days and evaluated the cell function. After the addition of RGE on the culture medium, we determined the effect of RGE on the cell viability, cell proliferation, protein synthesis, alkaline phosphatase activity, collagen synthesis and calcified nodule formation of the cultivated osteoblast. Results: The results were summarized as follows. 1. RGE did not change the survival rate of rat calvarial osteoblast. 2. RGE increased the proliferation of rat calvarial osteoblast. 3. RGE increased ALP activity of rat calvarial osteoblast., 4. RGE slightly affected protein synthesis of rat calvarial osteoblast. 5. RGE increased collagen synthesis of rat calvarial osteoblast. 6. RGE slightly affected calcified nodule formation of rat calvarial osteoblast. Conclusions: From these results, it is concluded that RG might improve the osteoporosis resulted from augmentation of osteoblast proliferation.

Quality Control of Ginseng Products(Part I) - The saponins isolated from ginseng roots and leaves - (인삼제품(人蔘製品)의 품질개량(品質改良)에 관(關)한 연구(硏究) (제일보(第一報)) - 인삼근(人蔘根) 및 엽(葉) Saponin의 비교연구(比較硏究) -)

  • Cho, Han-Ok;Cho, Sung-Hwan;Kim, Soo-Ja
    • Applied Biological Chemistry
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    • v.22 no.1
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    • pp.10-17
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    • 1979
  • The saponins isolated form the herb of Panax ginseng C.A. Meyer were investigated as compared with ginseng root saponins. By adopting DEAE cellulose ion exchange chromatography the pure saponins were isolated from Korean ginseng roots and leaves. The ginseng root and leaf saponins showed some differences in the pattern of the two-dimensional thin layer chromatogram. The ratio of panaxadiol to panaxatriol in the saponins was 1.7 in the roots and 3.5 in the leaves. Infra-red spectrum of ginseng leaf saponins isolated by liquid chromatography was identical with that of root saponins.

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Antioxidant and Hepatoprotective Effects of Hydroponic-cultured Ginseng Folium by fermentation (발효에 의한 수경재배 인삼 잎의 항산화 및 간 보호 효과)

  • Lee, Ah Reum;Park, Jae Ho
    • The Korea Journal of Herbology
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    • v.30 no.4
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    • pp.101-108
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    • 2015
  • Objectives : Positive effects of Ginseng has great research attentions such as anticancer, anti-diabetic, antiaging, liver, immune function, CNS, etc. In this study, we investigated Hydroponic-cultured Ginseng Folium fermented byBacillus subtilisto establish fermentation conditions for enhancing functionality.Methods : Ginseng Folium were cultivated hydroponic-cultured and were extracted with methanol. We inoculateBacillus subtilisfor fermentation by adding to 0%, 3% and 5% sugar respectively and checked antioxidant activities, total phenolic content and total saponin content in 2 days intervals during 11 days. The antioxidant activities were studied by the 1,1-diphenyl-2-picryl hydrazyl(DPPH) radical, 2, 2'-Azino-bis(3-ethylbenzothiazoline-6 sulfonic acid) diammonium salt(ABTS) radical scavenging assay and Reducing power assay. We analyzed the Total phenol content, crude saponin content and ginsenoside content. Moreever, Hepatoprotective effects by Glutamic oxaloacetic transaminase(GOT) and Glutamic pyruvic transaminase(GPT) in Sprague-Dawley rat.Results : The results of DPPH and ABTS were 66.89% and 96.72%, respectively. The reducing power was resulted in optical density of 0.7312 with 3% sugar after 9 days of fermentation. and the concentration at 200 ㎍/㎖. Total phenol content was 36.92㎎/g with 3% sugar after 9 days of fermentation, in which crude saponin content wasn't changed, and ginsenoside content such as Rg3, Re and Rb was increased. Activities of GOT and GPT concentration were decreased in rat.Conclusions : This study suggests that hydroponic-cultured Ginseng Folium fermented byBacillus subtilisin 9 days showed significant efficacy of hepato-protection as well as antioxidant compared to the others. In addition, it shows not only improved value but also utilized hydroponic-cultured Ginseng Folium by fermentation.