• Journal of Ginseng Research
• /
• 제39권3호
• /
• pp.221-229
• /
• 2015

Background: Minor ginsenosides, those having low content in ginseng, have higher pharmacological activities. To obtain minor ginsenosides, the biotransformation of American ginseng protopanaxadiol (PPD)-ginsenoside was studied using special ginsenosidase type-I from Aspergillus niger g.848. Methods: DEAE (diethylaminoethyl)-cellulose and polyacrylamide gel electrophoresis were used in enzyme purification, thin-layer chromatography and high performance liquid chromatography (HPLC) were used in enzyme hydrolysis and kinetics; crude enzyme was used in minor ginsenoside preparation from PPD-ginsenoside; the products were separated with silica-gel-column, and recognized by HPLC and NMR (Nuclear Magnetic Resonance). Results: The enzyme molecular weight was 75 kDa; the enzyme firstly hydrolyzed the C-20 position 20-O-${\beta}$-D-Glc of ginsenoside Rb1, then the C-3 position 3-O-${\beta}$-D-Glc with the pathway $Rb1{\rightarrow}Rd{\rightarrow}F2{\rightarrow}C-K$. However, the enzyme firstly hydrolyzed C-3 position 3-O-${\beta}$-D-Glc of ginsenoside Rb2 and Rc, finally hydrolyzed 20-O-L-Ara with the pathway $Rb2{\rightarrow}C-O{\rightarrow}C-Y{\rightarrow}C-K$, and $Rc{\rightarrow}C-Mc1{\rightarrow}C-Mc{\rightarrow}C-K$. According to enzyme kinetics, $K_m$ and $V_{max}$ of Michaelis-Menten equation, the enzyme reaction velocities on ginsenosides were Rb1 > Rb2 > Rc > Rd. However, the pure enzyme yield was only 3.1%, so crude enzyme was used for minor ginsenoside preparation. When the crude enzyme was reacted in 3% American ginseng PPD-ginsenoside (containing Rb1, Rb2, Rc, and Rd) at $45^{\circ}C$ and pH 5.0 for 18 h, the main products were minor ginsenosides C-Mc, C-Y, F2, and C-K; average molar yields were 43.7% for C-Mc from Rc, 42.4% for C-Y from Rb2, and 69.5% for F2 and C-K from Rb1 and Rd. Conclusion: Four monomer minor ginsenosides were successfully produced (at low-cost) from the PPD-ginsenosides using crude enzyme.

• Journal of Ginseng Research
• /
• 제44권2호
• /
• pp.215-221
• /
• 2020

Background: Panax ginseng has been used for a variety of medical purposes in eastern countries for more than two thousand years. From the extensive experiences accumulated in its long medication use history and the substantial strong evidence in modern research studies, we know that ginseng has various pharmacological activities, such as antitumor, antidiabetic, antioxidant, and cardiovascular system-protective effects. The active chemical constituents of ginseng, ginsenosides, are rich in structural diversity and exhibit a wide range of biological activities. Methods: Ginsenoside constituents from P. ginseng flower buds were isolated and purified by various chromatographic methods, and their structures were identified by spectroscopic analysis and comparison with the reported data. The 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H- tetrazolium bromide method was used to test their cytotoxic effects on three human cancer cell lines. Results: Six ginsenosides, namely 6'-malonyl formyl ginsenoside F₁ (1), 3β-acetoxyl ginsenoside F₁ (2), ginsenoside Rh₂₄ (6), ginsenoside Rh₂₅ (7), 7β-hydroxyl ginsenoside Rd (8) and ginsenoside Rh₂₆ (10) were isolated and elucidated as new compounds, together with four known compounds (3-5 and 9). In addition, the cytotoxicity of these isolated compounds was shown as half inhibitory concentration values, a tentative structure-activity relationship was also discussed based on the results of our bioassay. Conclusion: The study of chemical constituents was useful for the quality control of P. ginseng flower buds. The study on antitumor activities showed that new Compound 1 exhibited moderate cytotoxic activities against HL-60, MGC80-3 and Hep-G2 with half inhibitory concentration values of 16.74, 29.51 and 20.48 μM, respectively.

• Journal of Applied Biological Chemistry
• /
• 제61권4호
• /
• pp.371-377
• /
• 2018

인삼(Panax. ginseng) 열매를 80% MeOH 수용액으로 3회 반복 추출한 뒤, 감압 농축한 추출물을 EtOAc, n-BuOH과 $H_2O$ 층으로 계통 분획을 실시하였다. EtOAc분획에 대하여 $SiO_2$ 및 ODS column chromatography를 반복실시하여 5종의 ginsenoside 화합물을 분리 및 정제하였다. NMR, IR, FAB/MS 데이터를 해석하여, 각각 ginsenoside F1 (1), ginsenoside F2 (2), ginsenpside F3 (3), ginsenoside Ia (4) 및 notoginsenoside Fe (5)로 구조 동정 하였다. 화합물 2-5는 인삼열매에서는 이번에 처음 분리 보고되었다. 분리한 5종의 화합물을 인체 암세포주(HCT-116, SK-OV-3, HeLa, HepG2, SK-MEL-5)에 처리하여 세포독성을 측정하였다. 이 중 화합물 2, 4, 및 5가 인체 암세포주에 대해 세포독성을 저해시키는 것을 알 수 있었다. 화합물 2는 SK-MEL-5, HepG2, HeLa세포에서 $IC_{50}$값이 82.8, 86.8, $78.3{\mu}M$로 확인되었다. 화합물 4는 HCT-116, SK-MEL-5, SK-OV-3, HepG2, HeLa 세포에서 $IC_{50}$ 값이 24.5, 25.4, 26.3, 22.0, $24.9{\mu}M$로 확인되었다. 화합물 5는 SK-MEL-5 세포에서 $IC_{50}$ 값이 $81.7{\mu}M$로 확인되었다. 인삼 열매에서 분리한 화합물2, 4, 및 5가 암세포주에 대해 강한 세포독성을 나타내는 것을 확인하였으며, 이 화합물들은 공통적으로 3번 수산기에 glucopyranose를 가지고 있음을 확인하였다.

• Journal of Ginseng Research
• /
• 제43권4호
• /
• pp.635-644
• /
• 2019

Background: To increase the pharmacological effects of red ginseng (RG, the steamed root of Panax ginseng Meyer), RG products modified by heat process or fermentation have been developed. However, the antiallergic effects of RG and modified/fermented RG have not been simultaneously examined. Therefore, we examined the allergic rhinitis (AR)-inhibitory effects of water-extracted RG (wRG), 50% ethanol-extracted RG (eRG), and bifidobacteria-fermented eRG (fRG) in vivo. Methods: RBL-2H3 cells were stimulated with phorbol 12-myristate-13-acetate/A23187. Mice with AR were prepared by treatment with ovalbumin. Allergic markers IgE, tumor necrosis factor-${\alpha}$, interleukin (IL)-4, and IL-5 were assayed in the blood, bronchoalveolar lavage fluid, nasal mucosa, and colon using enzyme-linked immunosorbent assay. Mast cells, eosinophils, and Th2 cell populations were assayed using a flow cytometer. Results: RG products potently inhibited IL-4 expression in phorbol 12-myristate-13-acetate/A23187-stimulated RBL-2H3 cells. Of tested RG products, fRG most potently inhibited IL-4 expression. RG products also alleviated ovalbumin-induced AR in mice. Of these, fRG most potently reduced nasal allergy symptoms and blood IgE levels. fRG treatment also reduced IL-4 and IL-5 levels in bronchoalveolar lavage fluid, nasal mucosa, and reduced mast cells, eosinophils, and Th2 cell populations. Furthermore, treatment with fRG reduced IL-4, IL-5, and IL-13 levels in the colon and restored ovalbumin-suppressed Bacteroidetes and Actinobacteria populations and ovalbumin-induced Firmicutes population in gut microbiota. Treatment with ginsenoside Rd significantly alleviated ovalbumin-induced AR in mice. Conclusion: fRG and ginsenoside Rd may alleviate AR by suppressing IgE, IL-4, IL-5, and IL-13 expression and restoring the composition of gut microbiota.

• Journal of Ginseng Research
• /
• 제34권4호
• /
• pp.288-295
• /
• 2010

Ginsenosides are the principal components responsible for the pharmacological and biological activities of ginseng. Ginsenoside Rd was transformed into compound K using cell-free extracts of food microorganisms, with Lactobacillus pentosus DC101 isolated from kimchi (traditional Korean fermented food) used for this conversion. The optimum time for the conversion was about 72 h at a constant pH of 7.0 and an optimum temperature of about $30^{\circ}C$. The transformation products were identified by thin-layer chromatography and high-performance liquid chromatography, and their structures were assigned using nuclear magnetic resonance analysis. Generally, ginsenoside Rd was converted into ginsenoside F2 by 36 h post-reaction. Consequently, over 97% of ginsenoside Rd was decomposed and converted into compound K by 72 h post-reaction. The bioconversion pathway to produce compound K is as follows: ginsenoside Rd$\rightarrow$ginsenoside F2$\rightarrow$compound K.

• 식물조직배양학회지
• /
• 제27권6호
• /
• pp.461-468
• /
• 2000

인삼 (Panax ginseng C.A. Meyer)의 뿌리로부터 유도 인삼모상근을 이용한 무기이온 흡수과정의 상승 및 길항작용에서 phosphorus (P $O_{4}$$^{-}$)와 nitrogen source (N $H_{4}$$^{+}$, N $O_{3}$$^{-}$)의 증가는 조직 내에 $Mg^{2+}$ 및 F $e^{2+}$ 의 흡수를 증가시키는 원인으로 확인되었다 특히, ammonium의 농도를 4배 높일 경우 F $e^{2+}$의 흡수는 13 mg/L, C $u^{2+}$의 흡수는 0.32mg/L이 증가하여 각각 47.5%와 123.1%로 현저한 이온흡수의 상승효과를 초래하였다. 이러한 결과는 인삼포장에서 인산과 질소원의 과도한 시비는 생장을 다소 증가시키고 뿌리의 철이온과 2가 무기이온 흡수도 증가시킴을 알 수 있다 유리당의 함량은 유리당의 종류에 따라 인산과 질소원의 최적 농도가 다르게 나타났다. 반면에 ginsenoside함량은 인산과 질소원 모두 가장 낮은 농도에서 가장 높았다. 인산의 경우에는 1/2 MS배지의 농도 (0.62 mM)보다 낮은 0.31 mM에서 R $b_2$는 44.7%, Rc는 29.9%가 높았다. Ammonium은 1/2 MS 배지 (10.30 mM)보다 낮은 5.15 mM에서 R $b_2$와 Rc를 각각 21.7%와 31.9% 그리고 nitrate 역시 1/2 MS배지 (9.4mM)보다 낮은 4.70 mM 에서 각각 17.6%와 25.5%의 뚜렷한 함량증가를 나타냈다. 질소원에 따른 ginsenoside 함량 증가에서는 N $H_{4}$$^{+}$가 N $O_{3}$$^{-}$보다 다소 효과적이었다. 따라서 인삼모상근의 ginsenoside생산을 높이기 위해서는 인산과 질소원의 농도를 조절한 새로운 배지가 적합한 것으로 확인되었다.

• 대한약학회:학술대회논문집
• /
• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
• /
• pp.108.3-109
• /
• 2003

In the present study, we investigated the antitumor effects of Ginsenoside Rh2 and $\beta$-glucan using an experimental metastatic mouse model intravenously injected with B 16 melanoma F10 cells. Oral administration to various concentration of $\beta$-glucan (50mg/kg, l00mg/kg and 200mg/kg) dose-dependently reduced the lung-metastatic potential of metastatic BI6 melanoma F10 cells in syngenic mice. At same dose, Ginsenoside Rh2(50mg/kg) has more antitumor effect than $\beta$-glucan(50mg/kg). (omitted)

• Journal of Ginseng Research
• /
• 제36권1호
• /
• pp.86-92
• /
• 2012

The glioblastoma multiforme (GBM) is the most common malignant brain tumor in adults. Despite combination treatments of radiation and chemotherapy, the survival periods are very short. Therefore, this study was conducted to assess the potential of ginsenoside $F_2$ (F2) to treat GBM. In in vitro experiments with glioblastoma cells U373MG, F2 showed the cytotoxic effect with $IC_{50}$ of 50 ${\mu}g/mL$ through apoptosis, confirmed by DNA condensation and fragmentation. The cell population of cell cycle sub-G1 as indicative of apoptosis was also increased. In xenograft model in SD rats, F2 at dosage of 35 mg/kg weight was intravenously injected every two days. This reduced the tumor growth in magnetic resonance imaging images. The immunohistochemistry revealed that the anticancer activity might be mediated through inhibition of proliferation judged by Ki67 and apoptosis induced by activation of caspase-3 and -8. And the lowered expression of CD31 showed the reduction in blood vessel densities. The expression of matrix metalloproteinase-9 for invasion of cancer was also inhibited. The cell populations with cancer stem cell markers of CD133 and nestin were reduced. The results of this study suggested that F2 could be a new potential chemotherapeutic drug for GBM treatment by inhibiting the growth and invasion of cancer.

• 동의생리병리학회지
• /
• 제21권4호
• /
• pp.856-859
• /
• 2007

We have investigated the antimetastatic and antitumor effects of Ginsenoside Rh2 and ${\beta}-glucan$ unsing an experimental metastatic mouse model intravenously injected with B 16 melanoma F 10 cells. Animal groups are divided into six groups according to the dosage of drug administration and the kind of drugs. The groups are control, ${\beta}-glucan$ with 50, 100 and 200 mg/kg, Geinsenoside Rh2 50 mg/kg, and ${\beta}-glucan$ 50 mg/kg + Ginsenoside Rh2 50 mg/kg. Oral administration of various concentration of ${\beta}-glucan$( 50, 100, and 200 mg/kg) were reduced the lung- metastatics induced by metastatic B16 melanoma F 10 cells injection with a dose dependent manner in the syngenic mice. At same dosage group, Ginsenoside Rh2 (50 mg/kg) has more antimetastatic effect than the ${\beta}-glucan$(50 mg/kg). The highest antimetastatic effects was observed in the ${\beta}-glucan$ 50 mg/kg + Ginsenoside Rh2 50 mg/kg group and has a similar tendency in the anti-tumor effects, including decrease of the average tumor weight and increase of the average survival rate. There are no differences of the average tumor weights were apparent in the ${\beta}-glucan$ groups, however there were little decrease of the average tumor weight in Ginsenoside 50 mg/kg group and ${\beta}-glucan$ 50 mg/kg + Ginsenoside Rh2 50 mg/kg group than that of the control group. The rate of average survival rate in the ${\beta}-glucan$ 50 mg/kg + Ginsenoside Rh2 50 mg/kg group, ${\beta}-glucan$ 200 mg/kg, ${\beta}-glucan$ 100 mg/kg and ${\beta}-glucan$ 50 mg/kg, and Ginsenoside 50 mg/kg groups were highly in order. These data suggest that antimetastatic and antitumor effect of combination of Ginsenodide Rh2 and ${\beta}-glucan$ be the highest in this study.

• 생약학회지
• /
• 제47권1호
• /
• pp.73-78
• /
• 2016

The purpose of this study is to develop a new preparation process of ginseng (Panax ginseng) flower buds extracts featuring high concentration of ginsenosides Rg2, Rg3, Rg5, F4 and Rh1, red ginseng special components. Chemical transformation from ginseng saponin glycosides to prosapogenin was analyzed by the HPLC. Extracts of ginseng flower buds were processed under several treatment conditions of ultrasonication (at $100^{\circ}C$). The results showed that the quantity of ginsenoside Rg6 increased by over 8.8% at the 16 hours of ultrasonication. Ginseng flower buds ethanol extract compared with other process times. The result of UGF-16 indicates that the ultrasonication processed ginseng flower buds extracts (at $100^{\circ}C$) treated for 16 hours produced the highest amount of ginsenoside F4 (8.833%), Rg3 (2.230%), Rg5 (2.339%) and Rg2 (1.002%).