• Proceedings of the Ginseng society Conference
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• 1998.06a
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• pp.146-159
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• 1998

A new processed ginseng with fortified activity is developed. The process comprise with the heat treatment of fresh or white ginseng at higher temperature and pressure than those used for the preparation of red ginseng. This new processed ginseng showed 7 times higher antioxidant activity and more than 30 times stronger vasodilating activity than those shown in raw ginseng. Other activities found in the new processed ginseng include cancer chemoprevention, antinephrotoxic, and antineurotoxic activities. Less polar ginsenosides isolated from processed ginseng exhibited anti-platelet aggregation activity and anti-cancer activity. Many ginsenosides were isolated from this new processed ginseng, namely 20(S)-$Rg_3$,20(R)-$Rg_3$, $Rg_5$, $Rg_6$, $F_4$, $Rh_4$,20(S)-$Rg_3$,20(R)-$Rg_3$ and $Rg_4$. In addition to these known compounds, seven new ginsenosides, named as gisenoside $Rk_1$, $Rk_2$, $Rk_3$, $Rs_4$, $Rs_5$, $Rs_6$, and $Rs_7$ were isolated. The major constituents of new processed ginseng were 20(S)-$Rg_3$,20(R)-$Rg_3$, $Rk_1$ and $Rg_5$ which are minors in red ginseng. Since the chemical constituents and biological activities of this new processed ginseng are quite different from those of white or red ginseng, we designated it as $'$sun ginseng (仙蔘)$'$.s;$.

• Proceedings of the Ginseng society Conference
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• 2007.05a
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• pp.23-24
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• 2007

Purpose: 인삼이 항당뇨 활성을 가진다는 연구가 많은 연구자들에 의해 진행되었고, 이는 인삼의 구성 성분 중 ginsenoside에 기인한다는 보고가 있다. 본 연구는 ginsenoside의 항당뇨 작용기전을 in vitro에서 알아보고자 3T3-L1 지방세포에서 glucose uptake와 췌장 베타세포인 HIT-T15 세포에서 insulin 분비 효과를 확인하였다. 이를 위하여 인삼을 식초로 처리한 긴삼의 70% MeOH 분획으로부터 protopanaxadiol 계인 ginsenoside $Rb_2$, $Rg_3$ 그리고 protopanaxtriol 계인 $Rg_2$를 분리하여 본 실험에 사용하였다. Method: Ginsenoside $Rb_2$, $Rg_2$, $Rg_3$가 지방 세포에서 glucose uptake에 미치는 효과를 확인하기 위하여 3T3-L1 세포를 DMEM (Dulbecco's Modified Eagle's Medium) 배지에서 분화 유도시켰으며 3T3-L1 preadipocyte가 80% 정도 자라면 분화 유도 배지 (5% fetal bovine serum (FBS), 0.5 mM isobutylmethylxanthine (IBMX), 1 mM dexamethasone 그리고 $10{\mu}g/ml$ insulin가 포함된 DMEM)로 4일, $10{\mu}g/ml$ insulin가 포함된 DMEM으로 2일, FBS만 포함된 DMEM으로 2일 배양하여 총 8일 동안 분화를 유도하였다. 분화 유도된 3T3-L1 adipocytes 에 각각 $Rb_2$, $Rg_2$, $Rg_3$를 $20{\mu}M$로 처리하여 16시간 배양하여 low glucose DMEM에서 3시간 배양한 후에 $37^{\circ}C$에서 insulin 10 ng/ml 과 각각 $Rb_2$, $Rg_2$, $Rg_3$가 포함된 Krebs Ringer Hepes buffer(KRP buffer)에서 20분간 배양하였다. 2-deoxy-D-[$^3H$]-glucose를 넣고 10분 후에 차가운 PBS로 반응을 종결시켜 lysis buffer로 cell을 모은 후 scintillation counter를 이용하여 glucose를 측정하였다. Insulin 분비 효과는 HIT-T15 세포와 일차 배양한 흰쥐 소도세포(islets)를 사용하여 확인하였다. HIT-T15 세포는 24 well plate에 well 당 $2{\times}10^5$ 개씩 분주하여 24시간 동안 배양한 후 시료를 처리하였으며 소도 세포는 Sprague-Dawley rat의 췌장에 collagenase가 포함된 Hanks' Balanced Salt Solution(HBSS)을 주입하여 분리하고 islets을 얻었다. 분리한 소도세포를 $1{\sim}2$일 동안 배양하여 $Rb_2$, $Rg_2$, $Rg_3$가 각각 $20{\mu}M$의 농도로 첨가된 insulin 측정용 buffer인 Krebs-Ringer buffer (KRB+0.3% BSA, KRBB)에 $37^{\circ}C$에서 1시간 incubation 시킨 후 배양액으로 분비된 인슐린의 양을 측정하였다. 한편 ginsenoside의 인슐린 분비 촉진 기전을 알아보기 위한 실험에서는 ATP-sensitive $K^+$ channel opener인 diazoxide (0.5 mM)가 ginsenoside에 의해 촉진된 인슐린 분비를 억제하는지 살펴보았다. Result: glucose uptake assay 에서는 $Rg_2$가 가장 크게 glucose uptake를 증가시켰고 $Rb_2$, $Rg_3$는 그 활성이 크지 않았다. 한편 Insulin 분비 효과는 diol계인 $Rg_3$에서 용량 의존적으로 인슐린의 분비를 촉진시켰으며 $20{\mu}M$ 농도에서 대조군과 비교해 1.5배 이상의 분비 촉진 효과를 보였고 triol계인 $Rg_2$ 에서는 이러한 효과가 나타나지 않았다. $Rg_3$의 인슐린 분비 촉진 기전을 0.5 mM 의 diazoxide를 이용하여 확인한 결과 $Rg_3$에 의해 촉진된 인슐린 분비를 감소시켰다. 이로 미루어보아 $Rg_3$의 인슐린 분비 촉진 기전은 ATP-sensitive $K^+$ 채널의 봉쇄에 의한 것임을 확인할 수 있었다.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.12
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• pp.6191-6196
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• 2012

Aims and Background: Traditional chemotherapy strategies for human leukemia commonly use drugs based on cytotoxicity to eradicate cancer cells. One predicament is that substantial damage to normal tissues is likely to occur in the course of standard treatments. Obviously, it is urgent to explore therapies that can effectively eliminate malignant cells without affecting normal cells. Our previous studies indicated that ginsenoside $Rg_1$ ($Rg_1$), a major active pharmacological ingredient of ginseng, could delay normal hematopoietic stem cell senescence. However, whether $Rg_1$ can induce cancer cell senescence is still unclear. Methods: In the current study, human leukemia K562 cells were subjected to $Rg_1$ exposure. The optimal drug concentration and duration with K562 cells was obtained by MTT colorimetric test. Effects of $Rg_1$ on cell cycle were analyzed using flow cytometry and by SA-${\beta}$-Gal staining. Colony-forming ability was measured by colony-assay. Telomere lengths were assessed by Southern blotting and expression of senescence-associated proteins P21, P16 and RB by Western blotting. Ultrastructural morphology changes were observed by transmission electron microscopy. Results: K562 cells demonstrated a maximum proliferation inhibition rate with an $Rg_1$ concentration of $20{\mu}\;mol{\cdot}L^{-1}$ for 48h, the cells exhibiting dramatic morphological alterations including an enlarged and flat cellular morphology, larger mitochondria and increased number of lysosomes. Senescence associated-${\beta}$-galactosidase (SA-${\beta}$-Gal) activity was increased. K562 cells also had decreased ability for colony formation, and shortened telomere length as well as reduction of proliferating potential and arrestin $G_2$/M phase after $Rg_1$ interaction. The senescence associated proteins P21, P16 and RB were significantly up-regulated. Conclusion: Ginsenoside $Rg_1$ can induce a state of senescence in human leukemia K562 cells, which is associated with p21-Rb and p16-Rb pathways.

• Journal of Ginseng Research
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• v.43 no.4
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• pp.580-588
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• 2019

Background: Ginsenoside Rg1 has been shown to clear senescence-associated beta-galactosidase (SA-${\beta}$-gal) in cultured cells. It remains unknown whether Rg1 can influence SA-${\beta}$-gal in exercising human skeletal muscle. Methods: To examine SA-${\beta}$-gal change, 12 young men (age $21{\pm}0.2years$) were enrolled in a randomized double-blind placebo controlled crossover study, under two occasions: placebo (PLA) and Rg1 (5 mg) supplementations 1 h prior to a high-intensity cycling (70% $VO_{2max}$). Muscle samples were collected by multiple biopsies before and after cycling exercise (0 h and 3 h). To avoid potential effect of muscle biopsy on performance assessment, cycling time to exhaustion test (80% $VO_{2max}$) was conducted on another 12 participants (age $23{\pm}0.5years$) with the same experimental design. Results: No changes of SA-${\beta}$-gal were observed after cycling in the PLA trial. On the contrary, nine of the 12 participants showed complete elimination of SA-${\beta}$-gal in exercised muscle after cycling in the Rg1 trial (p < 0.05). Increases in apoptotic DNA fragmentation (PLA: +87% vs. Rg1: +133%, p < 0.05) and $CD68^+$ (PLA:+78% vs. Rg1:+121%, p = 0.17) occurred immediately after cycling in both trials. During the 3-h recovery, reverses in apoptotic nuclei content (PLA:+5% vs. Rg1 -32%, p < 0.01) and increases in inducible nitrate oxide synthase and interleukin 6 mRNA levels of exercised muscle were observed only in the Rg1 trial (p < 0.01). Conclusion: Rg1 supplementation effectively eliminates senescent cells in exercising human skeletal muscle and improves high-intensity endurance performance.

• Korean Journal of Food Science and Technology
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• v.44 no.4
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• pp.411-416
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• 2012

This study was carried out to investigate the effects of the extrusion process on the change of components in ginseng. The extraction yields from ginseng by distilled water extraction were highest in the extruded ginsengs, whereas it was lowest in the white ginseng. The contents of crude saponin were highest in the extruded ginseng, and they increased as the extrusion temperature was raised. The total contents of 11 kinds of ginsenosides increased in the order of red, white and extruded ginsengs. In particular, red ginseng showed higher contents of Rg1, Rg3 and Rb2, whereas Re was highest in white ginseng. In addition, the contents of Rg2, Rh1, Rh2 and Rg3 in the extruded white ginseng became higher. Free sugar contents were greatest in red ginseng. However, they were lowest in the extruded ginseng. White ginseng had a greater L value, whereas extruded ginseng demonstrated higher a and b values. In conclusion, the extraction yields, the contents of saponin, and ginsenoside-Rg2, Rh1, Rh2 and Rg3 were increased through the extrusion process.

• Journal of Ginseng Research
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• v.38 no.3
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• pp.194-202
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• 2014

Background: Ginsenosides, the major ingredients of Panax ginseng, have been studied for many decades in Asian countries as a result of their wide range of pharmacological properties. The less polar ginsenosides, with one or two sugar residues, are not present in nature and are produced during manufacturing processes by methods such as heating, steaming, acid hydrolysis, and enzyme reactions. $^1H$-NMR and $^{13}C$-NMR spectroscopic data for the identification of the less polar ginsenosides are often unavailable or incomplete. Methods: We isolated 21 compounds, including 10 pairs of 20(S) and 20(R) less polar ginsenosides (1-20), and an oleanane-type triterpene (21) from a processed ginseng preparation and obtained complete $^1H$-NMR and $^{13}C$-NMR spectroscopic data for the following compounds, referred to as compounds 1-21 for rapid identification: 20(S)-ginsenosides Rh2 (1), 20(R)-Rh2 (2), 20(S)-Rg3 (3), 20(R)-Rg3 (4), 6'-O-acetyl-20(S)-Rh2 [20(S)-AcetylRh2] (5), 20(R)-AcetylRh2 (6), 25-hydroxy-20(S)-Rh2 (7), 25-hydroxy-20(S)-Rh2 (8), 20(S)-Rh1 (9), 20(R)-Rh1 (10), 20(S)-Rg2 (11), 20(R)-Rg2 (12), 25-hydroxy-20(S)-Rh1 (13), 25-hydroxy-20(R)-Rh1 (14), 20(S)-AcetylRg2 (15), 20(R)-AcetylRg2 (16), Rh4 (17), Rg5 (18), Rk1 (19), 25-hydroxy-Rh4 (20), and oleanolic acid 28-O-b-D-glucopyranoside (21).

• Journal of Ginseng Research
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• v.27 no.3
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• pp.110-114
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• 2003

Objective; Antistress effect of Korean red ginseng (RG) on postmenopausal women with severe climacteric syndrome (CS) were evaluated from the viewpoint of traditional KAMPO-medicine and Western medicine. Methods; All patients with CS were treated with daily oral administration of 6g RG for 30 days. Nine patients with CS were evaluated with the use of diagnostic scores for KI-deficiency (deficiency of vital energy) and OKETSU (blood stagnation) syndrome from the viewpoint of KAMPO-medicine. In the same patients with CS, peripheral blood levels of ${\beta}$-endorphin and total plasminogen activator inhibitor-1 (t-PAI-1) were measured before and after treatment with RG. In another group, 12 patients with CS, psychological test using CMI, STAI and SDS were performed from the viewpoint of Western medicine. Stress related hormones, such as ACTH, cortisol and DHEA-S in those 12 patients with CS were also measured before and after treatment with RG. Results; KI-deficiency score and OKETSU score in patients with CS were significantly (p<0.001) higher than those in patients without CS. After treatment with RG, both scores were markedly (p<0.001) decreased compared to before treatment with RG. ${\beta}$-endorphin levels in patients with CS were significantly (p<0.05) higher than those in patients without CS. Total PAI-levels in patients with CS were increased before treatment with RG. No significant difference, however, were observed between patients with and without CS. After treatment with RG, both levels of ${\beta}$-endorphin and total PAI-1 in patients with CS were significantly (p<0.001 and p<0.05, respectively) decreased compared to before treatment with RG. CMI and STAI scores in patients with CS were significantly (p<0.05) higher than those in patients without CS. SDS scores in patients with CS were also markedly (p<0.00l) higher than in those without CS. After treatment with RG, all scores decreased within normal range. DHEA-S levels in patients with CS were about a half of those without CS. Consequently, cortisol/DHEA-S (C/D) ratio was significantly(p<0.001) higher in patients with CS than in those without CS. Although the decreased DHEA-S levels were not restored to the levels in patients without CS, the C/D ratio decreased significantly (p<0.05) after treatment with RG. Conclusion; Reinforcement of vital energy and improvement of stagnant blood circulations by oral administration of RG were elucidated from the viewpoint of traditional KAMPO-medicine. From the viewpoint of Western medicine, effect of RG on postmenopsusal women with CS seemed to be brought about in part by not only an improvement of psychoneuroendocrine dysfunctions but also an amelioration of blood coagulation systems.

• Journal of Ginseng Research
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• v.44 no.2
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• pp.341-349
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• 2020

Background: The replicative senescence of human dermal fibroblasts (HDFs) is accompanied by growth arrest. In our previous study, the treatment of senescent HDFs with Rg3(S) lowered the intrinsic reactive oxygen species (ROS) levels and reversed cellular senescence by inducing peroxiredoxin-3, an antioxidant enzyme. However, the signaling pathways involved in Rg3(S)-induced senescence reversal in HDFs and the relatedness of the stereoisomer Rg3(R) in corresponding signaling pathways are not known yet. Methods: We performed senescence-associated β-galactosidase and cell cycle assays in Rg3(S)-treated senescent HDFs. The levels of ROS, adenosine triphosphate (ATP), and cyclic adenosine monophosphate (cAMP) as well as the mitochondrial DNA copy number, nicotinamide adenine dinucleotide (NAD)⁺/1,4-dihydronicotinamide adenine dinucleotide (NADH) ratio, and NAD-dependent sirtuins expression were measured and compared among young, old, and Rg3(S)-pretreated old HDFs. Major signaling pathways of phosphatidylinositol 3-kinase/Akt, 5' adenosine monophosphate-activated protein kinase (AMPK), and sirtuin 1/3, including cell cycle regulatory proteins, were examined by immunoblot analysis. Results: Ginsenoside Rg3(S) reversed the replicative senescence of HDFs by restoring the ATP level and NAD⁺/NADH ratio in downregulated senescent HDFs. Rg3(S) recovered directly the cellular levels of ROS and the NAD⁺/NADH ratio in young HDFs inactivated by rotenone. Rg3(S) mainly downregulated phosphatidylinositol 3-kinase/Akt through the inhibition of mTOR by cell cycle regulators like p53/p21 in senescent HDFs, whereas Rg3(R) did not alter the corresponding signaling pathways. Rg3(S)-activated sirtuin 3/PGC1α to stimulate mitochondrial biogenesis. Conclusion: Cellular molecular analysis suggests that Rg3(S) specifically reverses the replicative senescence of HDFs by modulating Akt-mTOR-sirtuin signaling to promote the biogenesis of mitochondria.

• Korean Journal of Plant Resources
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• v.19 no.1
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• pp.139-143
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• 2006

To increase the contents of functional ginsenosides by conversion, especially ginsenoside-$Rg_3$ and $Rh_2$, the extracts of red ginseng were treated with high temperature and citric acid or apricot extract. When the extracts were subject to $120^{\circ}C$ for 2 hours, the content of ginsenoside-$Rg_3$ was increased 2 times than in control. When the extracts were subject to $120^{\circ}C$ and acidic conditions adjusted with citric acid, the ginsenoside-$Rg_3$, was detected 2.8 times, but other ginsenoside were decreased heavily to 65%. When the extract were treated with for 12 hours at $80^{\circ}C$, the content of ginsenoside-$Rg_3$ was increased to 3.3 times, Also, when the red ginseng extracts were treated with apricot extract, the ginsenoside-$Rg_3$ was detected to 4 times than in control, but other ginsenoside were decreased lightly to 35%, not same as at the $120^{\circ}C$ treatment.

• Journal of Ginseng Research
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• v.45 no.3
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• pp.433-441
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• 2021

Background: Multiple sclerosis (MS) and its animal model, the experimental autoimmune encephalomyelitis (EAE), are primarily characterized as dysfunction of the blood-brain barrier (BBB). Ginsenoside-Rg3-enriched Korean Red Ginseng extract (Rg3-KRGE) is known to exert neuroprotective, anti-inflammatory, and anti-oxidative effects on neurological disorders. However, effects of Rg3-KRGE in EAE remain unclear. Methods: Here, we investigated whether Rg3-KRGE may improve the symptoms and pathological features of myelin oligodendroglial glycoprotein (MOG)_35-55 peptide - induced chronic EAE mice through improving the integrity of the BBB. Results: Rg3-KRGE decreased EAE score and spinal demyelination. Rg3-KRGE inhibited Evan's blue dye leakage in spinal cord, suppressed increases of adhesion molecule platelet endothelial cell adhesion molecule-1, extracellular matrix proteins fibronection, and matrix metallopeptidase-9, and prevented decreases of tight junction proteins zonula occludens-1, claudin-3, and claudin-5 in spinal cord following EAE induction. Rg3-KRGE repressed increases of proinflammatory transcripts cyclooxygenase-2, inducible nitric oxide synthase, interleukin (IL)-1 beta, IL-6, and tumor necrosis factor-alpha, but enhanced expression levels of anti-inflammatory transcripts arginase-1 and IL-10 in the spinal cord following EAE induction. Rg3-KRGE inhibited the expression of oxidative stress markers (MitoSOX and 4-hydroxynonenal), the enhancement of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase 2 (NOX2) and NOX4, and NADPH activity in the spinal cord of chronic EAE mice. Furthermore, apocynin, a NOX inhibitor, mimicked beneficial effects of Rg3-KRGE in chronic EAE mice. Conclusion: Our findings suggest that Rg3-KRGE might alleviate behavioral symptoms and pathological features of MS by improving BBB integrity through modulation of NOX2/4 expression.