• Journal of Ginseng Research
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• 제40권1호
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• pp.47-54
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• 2016

Background: The roots of Panax ginseng contain noble tetracyclic triterpenoid saponins derived from dammarenediol-II. Dammarene-type ginsenosides are classified into the protopanaxadiol (PPD) and protopanaxatriol (PPT) groups based on their triterpene aglycone structures. Two cytochrome P450 (CYP) genes (CYP716A47 and CYP716A53v2) are critical for the production of PPD and PPT aglycones, respectively. CYP716A53v2 is a protopanaxadiol 6-hydroxylase that catalyzes PPT production from PPD in P. ginseng. Methods: We constructed transgenic P. ginseng lines overexpressing or silencing (via RNA interference) the CYP716A53v2 gene and analyzed changes in their ginsenoside profiles. Result: Overexpression of CYP716A53v2 led to increased accumulation of CYP716A53v2 mRNA in all transgenic roots compared to nontransgenic roots. Conversely, silencing of CYP716A53v2 mRNA in RNAi transgenic roots resulted in reduced CYP716A53v2 transcription. HPLC analysis revealed that transgenic roots overexpressing CYP716A53v2 contained higher levels of PPT-group ginsenosides ($Rg_1$, Re, and Rf) but lower levels of PPD-group ginsenosides (Rb1, Rc, $Rb_2$, and Rd). By contrast, RNAi transgenic roots contained lower levels of PPT-group compounds and higher levels of PPD-group compounds. Conclusion: The production of PPD- and PPT-group ginsenosides can be altered by changing the expression of CYP716A53v2 in transgenic P. ginseng. The biological activities of PPD-group ginsenosides are known to differ from those of the PPT group. Thus, increasing or decreasing the levels of PPT-group ginsenosides in transgenic P. ginseng may yield new medicinal uses for transgenic P. ginseng.

• Journal of Ginseng Research
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• 제41권1호
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• pp.10-22
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• 2017

Background: Asian ginseng and American ginseng are functional foods that share a close genetic relationship and are well-known worldwide. This article aims to investigate the correlation between morphological characteristics and the inherent quality of Asian and American ginsengs. Methods: In this study, an ultra-HPLC-quadrupole/time-of-flight MS (UHPLC-Q/TOF-MS) method was established for the quantitative analysis of 45 ginseng samples. The method developed for determination was precise and accurate. Results: The results showed that Asian ginseng samples with the same growing time (with the same or similar number of stem scars) that had a thinner main root, a longer rhizome and more branch roots contained greater amounts of ginsenosides. For American ginseng, two tendencies were observed in the relationship between the diameter of the main root and contents of ginsenosides. One tendency was that samples with thinner main roots tended to contain higher levels of ginsenosides, which was observed in the samples sold under the commercial name pao-shen. Another tendency was that samples with thicker main roots contained higher contents of ginsenosides, which was observed in the samples sold under the commercial name pao-mian, as well as in samples of American ginseng cultivated in Jilin, China. Conclusion: An approach using ultra-HPLC-quadrupole/time-of-flight MS was successfully established to link morphology and active components for evaluating the quality of Asian and American ginsengs. Clear correlation between visible morphological features and quality of Asian and American ginsengs was found. People can see the difference; this means consumers and vendors can evaluate ginseng by themselves.

• 분석과학
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• 제30권3호
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• pp.146-153
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• 2017

In this study, we isolated ginseng crude saponin (GCS) from Korean red ginseng (KRG) and determined the ginsenoside content in it to investigate the physiological and pathological effects of GCS on acute pulmonary inflammation induced by intratracheal instillation of cigarette smoke condensates (CSC) and lipopolysaccharide (LPS) solution in BALB/c mice. GCS was orally administered at doses of 10 mg/kg and 25 mg/kg for 3 weeks. The recovery rate of GCS from KRG was 6.5 % and total ginsenosides from GCS was 1.13 %, and the content of Rb1 was the highest among them. Total inflammatory cells in the lung homogenates and bronchoalveolar lavage fluid (BALF) increased following intratracheal administration of CSC and LPS. However, GCS administration impaired this increase. Furthermore, it inhibited the increase in leukocytes in the blood, considerably decreased neutrophils in BALF, and declined infiltration of inflammatory cells and deposition of collagen in the tracheal and alveolar tissue. In this study, GCS was found to have a protective effect against acute pulmonary inflammation and it may be beneficial in preventing various respiratory diseases.

• Journal of Ginseng Research
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• 제36권3호
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• pp.277-290
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• 2012

The metabolic profiles of Panax quinquefolius and its associated therapeutic values are critically affected by the repetitious steaming times. The times-dependent steaming effect of P. quinquefolius is not well-characterized and there is also no official guideline on its times of steaming. In this paper, a UPLC-Q-TOF-MS/MS method was developed for the qualitative profiling of multi-parametric metabolic changes of raw P. quinquefolius during the repetitious steaming process. Our method was successful in discriminating the differentially multi-steamed herbs. Meantime, the repetitious steaming-inducing chemical transformations in the preparation of black American ginseng (American ginseng that was subjected to 9 cycles of steaming treatment) were evaluated by this UPLC-Q-TOF-MS/MS based chemical profiling method. Under the optimized UPLC-Q-TOF-MS/MS conditions, 29 major ginsenosides were unambiguously identified and/or tentatively assigned in both raw and multi-steamed P. quinquefolius within 19 min, among them 18 ginsenosides were detected to be newly generated during the preparatory process of black American ginseng. The mechanisms involved were further deduced to be hydrolysis, dehydration, decarboxylation and addition reactions of the original ginsenosides in raw P. quinquefolius through analyzing mimic 9 cycles of steaming extracts of 14 pure reference ginsenosides. Our novel steaming times-dependent metabolic profiling approach represents the paradigm shift in the global quality control of multi-steamed P. quinquefolius products.

• Journal of Ginseng Research
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• 제41권3호
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• pp.379-385
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• 2017

In this study, white ginseng was used as the raw material, which was fermented with Paecilomyces hepiali through solid culture medium, to produce ginsenosides with modified chemical composition. The characteristic chemical markers of the products thus produced were investigated using rapid resolution liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (RRLC-QTOF-MS). Chemical profiling data were obtained, which were then subjected to multivariate statistical analysis for the systematic comparison of active ingredients in white ginseng and fermented ginseng to understand the beneficial properties of ginsenoside metabolites. In addition, the effects of these components on biological activity were investigated to understand the improvements in the phagocytic function of macrophages in zebrafish. According to the established RRLC-QTOF-MS chemical profiling, the contents in ginsenosides of high molecular weight, especially malonylated protopanaxadiol ginsenosides, were slightly reduced due to the fermentation, which were hydrolyzed into rare and minor ginsenosides. Moreover, the facilitation of macrophage phagocytic function in zebrafish following treatment with different ginseng extracts confirmed that the fermented ginseng is superior to white ginseng. Our results prove that there is a profound change in chemical constituents of ginsenosides during the fermentation process, which has a significant effect on the biological activity of these compounds.

• Journal of Ginseng Research
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• 제43권3호
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• pp.488-495
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• 2019

Background: Timosaponin AIII (TA3) is a steroidal saponin extracted from Anemarrhena asphodeloides. Here, we investigated the anticancer effects of TA3 in MG63 human osteosarcoma cells. TA3 attenuates migration and invasion of MG63 cells via regulations of two matrix metalloproteinases (MMPs), MMP-2 and MMP-9, which are involved with cancer metastasis in various cancer cells. TA3 reduced enzymatic activities and transcriptional expressions of MMP-2 and MMP-9 in MG63 cells. TA3 also inhibited Src, focal adhesion kinase, extracellular signal-regulated kinase (ERK1/2), c-Jun N-terminal kinase (JNK), p38, ${\beta}-catenin$, and cAMP response element binding signaling, which regulate migration and invasion of cells. TA3 induced apoptosis of MG63 cells via regulations of caspase-3, caspase-7, and poly(ADP-ribose) polymerase (PARP). Then, we tested several ginsenosides to be used in combination with TA3 for the synergistic anticancer effects. We found that ginsenosides Rb1 and Rc have synergistic effects on TA3-induced apoptosis in MG63 cells. Methods: We investigated the anticancer effects of TA3 and synergistic effects of various ginseng saponins on TA3-induced apoptosis in MG63 cells. To test antimetastatic effects, we performed wound healing migration assay, Boyden chamber invasion assays, gelatin zymography assay, and Western blot analysis. Annexin V/PI staining apoptosis assay was performed to determine the apoptotic effect of TA3 and ginsenosides. Results: TA3 attenuated migration and invasion of MG63 cells and induced apoptosis of MG63 cells. Ginsenosides Rb1 and Rc showed the synergistic effects on TA3-induced apoptosis in MG63 cells. Conclusions: The results strongly suggest that the combination of TA3 and the two ginsenosides Rb1 and Rc may be a strong candidate for the effective antiosteosarcoma agent.

• Food Science and Biotechnology
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• 제18권2호
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• pp.565-569
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• 2009

This study was conducted to evaluate the effects of different preparation methods on the recovery and quantification of ginsenosides in raw Korean ginseng (Panax ginseng C.A. Meyer). Eight major ginsenosides ($Rb_1$, $Rb_2$, $Rb_3$, Rc, Rd, Re, Rf, and $Rg_1$) were analyzed by high performance liquid chromatography (HPLC), after which the recovery and repeatability of the extraction of those ginsenosides using 3 different preparation methods were compared [A. direct extraction (DE) method, hot MeOH extraction/evaporation/direct dissolution; B. solid phase extraction (SPE) method, hot MeOH extraction/evaporation/dissolution/$C_{18}$ cartridge adsorption/MeOH elution; C. liquid-liquid extraction (LLE) method, hot MeOH extraction/evaporation/dissolution/n-BuOH fractionation]. Use of the DE method resulted in a significantly higher recovery of total ginsenosides than other methods and a relatively clear peak resolution. Use of the SPE and LLE methods resulted in clearer peak resolution, but lower ginsenoside recovery than the DE method. The LLE method showed the lowest ginsenoside recovery and repeatability among the 3 methods. Given that the DE method employed only extraction, evaporation, and a dissolution step (avoiding complicate and time consuming purification), this technique may be an effective method for the preparation and quantification of ginsenosides from raw Korean ginseng.

• 한국약용작물학회지
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• 제24권1호
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• pp.47-54
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• 2016

Background : A new extraction method-heated ultrasonic extraction was qualitatively and quantitatively analyzed for the extraction of major ginsenosides from ginseng extract; this new high-performance liquid chromatography (HPLC) method was compared with the official extraction method of Korean industrial standards and standard for health functional food. Methods and Results : Ginsenoside compounds were analyzed for 35 minutes by the new HPLC analysis method using a Halo$^{(R)}$ RP-Amide column. The new HPLC analysis method was validated by the measurement of intra-day and inter-day precision, accuracy, limit of detection (LOD), and limit of quantification (LOQ) of each ginsenoside. The correlation coefficients (r2) for the calibration curves of the ginsenoside compounds were over 0.9997 in terms of linearity. The heated ultrasonic extraction method using ultrasonication for 30 minutes at $50^{\circ}C$ yielded higher amount of ginsenosides than the extraction method of the Korean industrial standards owing to the enhancement of extraction efficiency. Conclusions : Compared to the other extraction methods, the heated ultrasonic extraction method yielded a higher amount of ginsenoside Rb1 than Rg1 index compounds for the quality evaluation of ginseng roots.

• Journal of Ginseng Research
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• 제41권4호
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• pp.487-495
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• 2017

Background: Although flowers of Panax ginseng Meyer (FPG), Panax quinquefolius L. (FPQ), and Panax notoginseng Burk. (FPN) have been historically used as both medicine and food, each is used differently in practice. Methods: To investigate the connection between components and enhancing immunity activity of FPG, FPQ, and FPN, a method based on a rapid LC coupled with quadrupole time-of-flight MS and immunomodulatory activity study evaluated by a carbon clearance test were combined. Results: According to quantitative results, the ratio of the total content of protopanaxatiol-type ginsenosides to protopanaxadiol-type ginsenosides in FPN was 0, but ranged from 1.10 to 1.32 and from 0.23 to 0.35 in FPG and FPQ, respectively. The ratio of the total content of neutral ginsenosides to the corresponding malonyl-ginsenosides in FPN ($5.52{\pm}1.33%$) was higher than FPG ($3.2{\pm}0.64%$) and FPQ ($2.39{\pm}0.57%$). The colorimetric analysis showed the content of total ginsenosides in FPQ, FPG, and FPN to be $13.75{\pm}0.60%$, $17.45{\pm}0.42%$, and $12.45{\pm}1.77%$, respectively. The carbon clearance assay indicated that the phagocytic activity of FPG and FPQ was higher than that of FPN. A clear discrimination among FPG, FPQ, and FPN was observed in the principal component analysis score plots. Seven compounds were confirmed to contribute strongly by loading plots, which may be the cause of differences in efficacy. Conclusion: This study provides basic information about the chemical and bioactive comparison of FPG, FPQ, and FPN, indicating that protopanaxtriol-type ginsenosides and malonyl-ginsenosides may play a key role in their enhancing immunity properties.

• 한국응용과학기술학회지
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• 제35권4호
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• pp.995-1002
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• 2018

Hairy root culture of ginseng is industrially prospected because the cultivation period of ginseng is relatively long. In this study, the effect of medium concentration and sucrose concentration on hairy root culture of ginseng was evaluated. The optimization of ginseng hairy roots transformed by Agrobacterium rhizogene were performed liquid medium. The MS(Murashinge & Skoog basal medium) concentration was selected with 1/2 strength MS and the optimal sucrose concentration was determined at 2-3%(w/v). At the optimum culture condition, The yield (the ratio of weight of grown hairy root cultures to weight of fresh ginseng hairy roots) and production rate of ginseng root were 19.42 times and 5.73 g/l-day. The major ginsenosides were Rb group, Re and Rg1. The produced total ginsenoside content in the solid medium was 9.87 (mg/g) and increased 1.34 times in the liquid medium (13.23 mg/g). In solid culture, the contents of ginsenosides Rb, Re and Rg1 were 2.14, 3.65 and 1.87 mg/g, respectively. In liquid culture, the contents of ginsenosides Rb, Re and Rg1 were 3.54, 4.12 and 2.63 mg/g, respectively.