• Journal of Ginseng Research
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• v.40 no.4
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• pp.400-408
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• 2016

Background: Ginsenoside Rh2 (GRh2) is the main bioactive component in American ginseng, a commonly used herb, and its antitumor activity had been studied in previous studies. PDZ-binding kinase/T-LAK cell-originated protein kinase (PBK/TOPK), a serine/threonine protein kinase, is highly expressed in HCT116 colorectal cancer cells. Methods: We examined the effect of GRh2 on HCT116 cells ex vivo. Next, we performed in vitro binding assay and in vitro kinase assay to search for the target of GRh2. Furthermore, we elucidated the underlying molecular mechanisms for the antitumor effect of GRh2 ex vivo and in vivo. Results: The results of our in vitro studies indicated that GRh2 can directly bind with PBK/TOPK and GRh2 also can directly inhibit PBK/TOPK activity. Ex vivo studies showed that GRh2 significantly induced cell death in HCT116 colorectal cancer cells. Further mechanistic study demonstrated that these compounds inhibited the phosphorylation levels of the extracellular regulated protein kinases 1/2 (ERK1/2) and (H3) in HCT116 colorectal cancer cells. In vivo studies showed GRh2 inhibited the growth of xenograft tumors of HCT116 cells and inhibited the phosphorylation levels of the extracellular regulated protein kinases 1/2 and histone H3. Conclusion: The results indicate that GRh2 exerts promising antitumor effect that is specific to human HCT116 colorectal cancer cells through inhibiting the activity of PBK/TOPK.

• Journal of Ginseng Research
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• v.39 no.4
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• pp.322-330
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• 2015

Background: UV-irradiated keratinocytes secrete various proinflammatory cytokines. UV-induced skin damage is mediated by growth factors and proinflammatory cytokines such as granulocyte macrophage colony stimulating factor (GM-CSF). In a previous study, we found that the saponin of Korean Red Ginseng (SKRG) decreased the expression of GM-CSF in UVB-irradiated SP-1 keratinocytes. In this study, we attempted to find the inhibitory mechanism of SKRG on UVB-induced GM-CSF expression in SP-1 keratinocytes. Methods: We investigated the inhibitory mechanism of SKRG and ginsenosides from Panax ginseng on UVB-induced GM-CSF expression in SP-1 keratinocytes. Results: Treatment with SKRG decreased the expression of GM-CSF mRNA and protein induced by irradiation of UVB in SP-1 keratinocytes. The phosphorylation of ERK was induced by UVB at 10 min, and decreased with SKRG treatment in SP-1 keratinocytes. In addition, treatment with SKRG inhibited the UVB-induced phosphorylation of epidermal growth factor receptor (EGFR), which is known to be an upstream signal of ERK. From these results, we found that the inhibition of GM-CSF expression by SKRG was derived from the decreased phosphorylation of EGFR. To identify the specific compound composing SKRG, we tested fifteen kinds of ginsenosides. Among these compounds, ginsenoside-Rh3 decreased the expression of GM-CSF protein and mRNA in SP-1 keratinocytes. Conclusion: Taken together, we found that treatment with SKRG decreased the phosphorylation of EGFR and ERK in UVB-irradiated SP-1 keratinocytes and subsequently inhibited the expression of GM-CSF. Furthermore, we identified ginsenoside-Rh3 as the active saponin in Korean Red Ginseng.

• Journal of Ginseng Research
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• v.33 no.4
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• pp.324-329
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• 2009

Cell-cell adhesion managed by various adhesion molecules is known to be one of pathophysiological phenomena found in numerous immunological diseases such as rheumatoid arthritis and allergic diseases. In this study, we examined the regulatory role of ginsenosides (G)- Rh1, reported to display anti-inflammatory and anti-allergic effects, on CD29-mediated cell adhesion. G-Rh1 significantly suppressed U937 cell-cell adhesion mediated by CD29 but not CD43. It also blocked U937 cell-fibronectin adhesion, mediated by activated CD29, up to 30%. In agreement, this compound also significantly decreased the surface level of CD29 but not CD43 as well as other costimulatory molecules such as CD69, CD80, and CD86. Therefore, these results suggest that G-Rh1 may have inhibitory function on CD29-mediated cell adhesion events, probably contributing to its anti-inflammatory and anti-allergic activities.

• Journal of Pharmacopuncture
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• v.19 no.3
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• pp.213-224
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• 2016

Objectives: Ginseng Rh2+ is enzyme-treated ginseng extract containing high amounts of converted ginsenosides, such as compound k, Rh2, Rg3, which have potent anticancer activity. We conducted general and genetic toxicity tests to evaluate the safety of ginseng Rh2+. Methods: An acute oral toxicity test was performed at a high-level dose of 4,000 mg/kg/day in Sprague-Dawley (SD) rats. A 14-day range-finding study was also conducted to set dose levels for the 90-day study. A subchronic 90-day toxicity study was performed at dose levels of 1,000 and 2,000 mg/kg/day to investigate the no-observed-adverse-effect level (NOAEL) of ginseng Rh2+ and target organs. To identify the mutagenic potential of ginseng Rh2+, we conducted a bacterial reverse mutation test (Ames test) using amino-acid-requiring strains of Salmonella typhimurium and Escherichia coli (E. coli), a chromosome aberration test with Chinese hamster lung (CHL) cells, and an in vivo micronucleus test using ICR mice bone marrow as recommended by the Korean Ministry of Food and Drug Safety. Results: According to the results of the acute oral toxicity study, the approximate lethal dose (ALD) of ginseng Rh2+ was estimated to be higher than 4,000 mg/kg. For the 90-day study, no toxicological effect of ginseng Rh2+ was observed in body-weight changes, food consumption, clinical signs, organ weights, histopathology, ophthalmology, and clinical pathology. The NOAEL of ginseng Rh2+ was established to be 2,000 mg/kg/day, and no target organ was found in this test. In addition, no evidence of mutagenicity was found either on the in vitro genotoxicity tests, including the Ames test and the chromosome aberration test, or on the in vivo in mice bone marrow micronucleus test. Conclusion: On the basis of our findings, ginseng Rh2+ is a non-toxic material with no genotoxicity. We expect that ginseng Rh2+ may be used as a novel adjuvant anticancer agent that is safe for long-term administration.

• Journal of Ginseng Research
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• v.44 no.5
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• pp.704-716
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• 2020

Background: Ginsenoside Rh2 (GRh2) is a characterized component in red ginseng widely used in Korea and China. GRh2 exhibits a wide range of pharmacological activities, such as anti-inflammatory, antioxidant, and anticancer properties. However, its effects on Toxoplasma gondii (T. gondii) infection have not been clarified yet. Methods: The effect of GRh2 against T. gondii was assessed under in vitro and in vivo experiments. The BV2 cells were infected with tachyzoites of T. gondii RH strain, and the effects of GRh2 were evaluated by MTT assay, morphological observations, immunofluorescence staining, a trypan blue exclusion assay, reverse transcription PCR, and Western blot analyses. The in vivo experiment was conducted with BALB/c mice inoculated with lethal amounts of tachyzoites with or without GRh2 treatment. Results and conclusion: The GRh2 treatment significantly inhibited the proliferation of T. gondii under in vitro and in vivo studies. Furthermore, GRh2 blocked the activation of microglia and specifically decreased the release of inflammatory mediators in response to T. gondii infection through TLR4/NF-κB signaling pathway. In mice, GRh2 conferred modest protection from a lethal dose of T. gondii. After the treatment, the proliferation of tachyzoites in the peritoneal cavity of infected mice markedly decreased. Moreover, GRh2 also significantly decreased the T. gondii burden in mouse brain tissues. These findings indicate that GRh2 exhibits an antieT. gondii effect and inhibits the microglial activation through TLR4/NF-κB signaling pathway, providing the basic pharmacological basis for the development of new drugs to treat toxoplasmic encephalitis.

• Korean Journal of Food Science and Technology
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• v.39 no.5
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• pp.494-499
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• 2007

The quality and functional properties of red ginseng in relation to steaming and drying conditions were evaluated. Fresh ginseng (5-year roots), cultivated in the Punggi region, were steamed for 2.5, 3.5, or 4.5 hr, and then dried by hot-air (60-$65^{\circ}C$/24 hr and $40^{\circ}C$,/3-4d) freezing ($-80^{\circ}C$/56 hr), and infrared (900 W/$62^{\circ}C$/68 hr). Hunter#s yellowness (b-value) and browning indexes (420 nm) of the samples were higher in the rootlets than in the main roots. Furthermore, these same index values were found to be high in the order of 3.5, 4.5, and 2.5 hr and infrared, hot-air, and freezing for steaming and subsequent drying, respectively. Analysis of soluble solids, total phenolics, total flavonoids, acidic polysaccharides, and electron donating abilities of the steamed and dried samples showed that 3.5hr of steaming with infrared drying was optimal. However, crude saponin contents were not influenced by steaming and drying conditions. The contents of $ginsenoside-Rg_l$, -Re, -Rf and $-Rb_2$, which were the major components in the samples, were reduced with steaming time, while the amounts of $-Rg_3$ and $-Rh_2$ increased, reaching the highest levels at 3.5 and 4.5 hr in the main roots and rootlets, respectively. The contents of $-Rg_3$ and $-Rh_2$ were similar in both the freeze-dried and hot-air dried samples.

• Korean Journal of Food Science and Technology
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• v.38 no.4
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• pp.521-525
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• 2006

Korean ginseng was heat treated at various temperatures (110, 120, 130, 140 and $150^{\circ}C$) and times (1, 2, 3, 4, and 5 hr). The heat treated ginseng extract was analyzed for the total polyphenol content, total flavonoid content, DPPH free-radical scavenging, 5-HMF and ginsenoside. The total phenolics and flavonoid content increased with increasing treatment temperature and time. The highest total phenolics content was 29.46 mg/g (d.b) in $150^{\circ}C$ for 1hr (control: 2.68 mg/g). The highest total flavonoid content was 4.75mg/g (d.b) in $150^{\circ}C$ for 2hr (control: 0.39 mg/g). The antioxidant activity increased until $140^{\circ}C$ for 3 hours. An extension of the treatment time did not have any effect, and the antioxidant activity decreased at temperatures higher than $150^{\circ}C$ for more than 2 hours. The content of ginsenoside $Rg_1$, Re, $Rb_2$ and Rb3 rapidly decreased with increasing treatment temperature and time. Ginsenoside $Rg_3$ and $Rh_2$ were newly produced, or their contents increased with increasing treatment temperature and time.

• Journal of Ginseng Research
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• v.19 no.3
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• pp.254-259
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• 1995

The content and composition of saponin compounds of Panax species were analyzed according to their species, region and processing type of red and white ginseng. The species employed were Korean-, Chinese-, Japanese red ginsengs, and Korean white ginseng of Panax ginseng, American- and Canadian ginsengs of Panax quinquefolium, and Panax notoinseng. Twelve main saponin components in the ginseng were identified and quantified using TLC and HPLC. All three species had remarkably different content and composition. However, within each species they were similar. Twelve major ginsenosides were determined in P. ginseng, eight in p. quinquefolium, and six in P. notoginseng. Of the components of P ginseng Rf, $Rh_1$, $Rh_2$ and Ra were not detected in P quinquefolium, and $Rb_2$, Rc, Rf, $Rh_2$, Ra and Ro not detected in P. notoinseam. Crude saponin content and protopanaxadiol/protopanaxatriol saponin ratio were compared. They were 4.81~5.24% and 1.27~ 1.45 in p. ginsengs, 7.01~7.25% and 2.12~ 2.15 in p. quinquefolium, 9.80% and 0.99 in P. notoineng. The prosapogenin and sapogenin content were different among the Panax species.

• Journal of Ginseng Research
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• v.45 no.2
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• pp.295-304
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• 2021

Background: Acute promyelocytic leukemia (APL) is a hematopoietic malignancy driven by promyelocytic leukemia-retinoic acid receptor A (PML-RARA) fusion gene. The therapeutic drugs currently used to treat APL have adverse effects. 20(S)-ginsenoside Rh2 (GRh2) is an anticancer medicine with high effectiveness and low toxicity. However, the underlying anticancer mechanisms of GRh2-induced PML-RARA degradation and apoptosis in human APL cell line (NB4 cells) remain unclear. Methods: Apoptosis-related indicators and PML-RARA expression were determined to investigate the effect of GRh2 on NB4 cells. Z-VAD-FMK, LY294002, and C 87, as inhibitors of caspase, and the phosphatidylinositol 3-kinase (PI3K) and tumor necrosis factor-α (TNF-α) pathways were used to clarify the relationship between GRh2-induced apoptosis and PML-RARA degradation. Results: GRh2 dose- and time-dependently decreased NB4 cell viability. GRh2-induced apoptosis, cell cycle arrest, and caspase3, caspase8, and caspase9 activation in NB4 cells after a 12-hour treatment. GRh2-induced apoptosis in NB4 cells was accompanied by massive production of reactive oxygen species, mitochondrial damage and upregulated Bax/Bcl-2 expression. GRh2 also induced PML/PML-RARA degradation, PML nuclear bodies formation, and activation of the downstream p53 pathway in NB4 cells. Z-VAD-FMK inhibited caspase activation and significantly reversed GRh2-induced apoptosis and PML-RARA degradation. GRh2 also upregulated TNF-α expression and inhibited Akt phosphorylation. LY294002, an inhibitor of the PI3K pathway, enhanced the antitumor effects of GRh2, and C 87, an inhibitor of the TNF-α pathway, reversed NB4 cell viability, and GRh2-mediated apoptosis in a caspase-8-dependent manner. Conclusion: GRh2 induced caspase-dependent PML-RARA degradation and apoptosis in NB4 cells via the Akt/Bax/caspase9 and TNF-α/caspase8 pathways.

• Korean Journal of Pharmacognosy
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• v.47 no.1
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• pp.73-78
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• 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%).