• Journal of Ginseng Research
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• v.41 no.3
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• pp.233-239
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• 2017

Background: Nephrotoxicity is the major side effect in cisplatin chemotherapy. Previously, we reported that the ginsenosides Rk3 and Rh4 reduced cisplatin toxicity on porcine renal proximal epithelial tubular cells (LLC-PK1). Here, we aimed to evaluate the protective effect of ginsenosides Rk3 and Rh4 on kidney function and elucidate their antioxidant effect using in vitro and in vivo models of cisplatin-induced acute renal failure. Methods: An enriched mixture of ginsenosides Rk3 and Rh4 (KG-KH; 49.3% and 43.1%, respectively) was purified from sun ginseng (heat processed Panax ginseng). Cytotoxicity was induced by treatment of $20{\mu}M$ cisplatin to LLC-PK1 cells and rat model of acute renal failure was generated by single intraperitoneal injection of 5 mg/kg cisplatin. Protective effects were assessed by determining cell viability, reactive oxygen species generation, blood urea nitrogen, serum creatinine, antioxidant enzyme activity, and histopathological examination. Results: The in vitro assay demonstrated that KG-KH ($50{\mu}g/mL$) significantly increased cell viability (4.6-fold), superoxide dismutase activity (2.8-fold), and glutathione reductase activity (1.5-fold), but reduced reactive oxygen species generation (56%) compared to cisplatin control cells. KG-KH (6 mg/kg, per os) also significantly inhibited renal edema (87% kidney index) and dysfunction (71.4% blood urea nitrogen, 67.4% creatinine) compared to cisplatin control rats. Of note, KG-KH significantly recovered the kidney levels of catalase (1.2-fold) and superoxide dismutase (1.5-fold). Conclusion: Considering the oxidative injury as an early trigger of cisplatin nephrotoxicity, our findings suggest that ginsenosides Rk3 and Rh4 protect the kidney from cisplatin-induced oxidative injury and help to recover renal function by restoring intrinsic antioxidant defenses.

• Journal of Applied Biological Chemistry
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• v.50 no.2
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• pp.78-84
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• 2007

The anti-fibrotic effects of hot water extract of adventitious root culture of Panax ginseng (ARCP) and the possible mechanisms were investigated on $CCl_4-induced$ hepatotoxicity model mice. Fibrosis was induced by a mild treatment of $CCl_4$. Then silymarin as a positive control drug and ARCP or carrier alone as a negative control were treated. Serum GPT, GOT and ALP activity levels were lowered by 25, 21 and 11% for silymarin treated group and by 48, 39 and 14% for ARCP treated group compared to carrier treated alone. Hepatic collagen for ARCP treatment group was reduced by 18% and MDA contents decreased a little more. Pro-fibrotic gene ($TGF-{\beta}1$, TIMP-1 and ${\alpha}-SMA$) expression increased following the $CCl_4$ treatment, but both the silymarin and the ARCP treatments decreased the expressions of these genes by 20% to 50%. The antioxidant effect of ARCP was studied by DPPH free radical scavenging activity. In addition, a generation of reactive oxygen species (ROS) was also reduced in $H_2O_2-treated$ HepG2 cells upon the ARCP treatment. In summary, ARCP has antioxidant property, and can have some protection against oxidative stress; more importantly, ARCP can efficiently protect mice against $CCl_4-induced$ fibrosis.

• Korean Journal of Medicinal Crop Science
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• v.28 no.3
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• pp.183-194
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• 2020

Background: This study investigated the effect of soil properties and soil bacterial community on early growth characteristics of wild-simulated ginseng (Panax ginseng C. A. Meyer) in coniferous and mixed forest experimental fields. Methods and Results: The soil bacterial community was analyzed using a high throughput sequencing technique (Illumina MiSeq sequencing). The relationship between the soil bacterial community, soil properties, and growth characteristics of wild-simulated ginseng were analyzed using principal coordinate analysis (PCoA) and the Pearson's correlation analysis. Soil properties and soil bacterial community showed significant difference with forest physiognomy. Results of Pearson's correlation analysis and PCoA showed that the soil properties (soil pH, organic matter, total nitrogen, and cation exchange capacity) and soil bacterial community had significant correlation with tree species ratio and early growth characteristics of wild-simulated ginseng. Conclusions: This study clearly demonstrated the effect of soil properties and soil bacterial community on early growth characteristics of wild-simulated ginseng in coniferous and mixed forest. Moreover, these results will help in the selection of suitable cultivation sites for wild-simulated ginseng.

• Journal of Korean Society of Forest Science
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• v.108 no.4
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• pp.522-530
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• 2019

Two edible plants, Allium victorialis var. platyphyllum and Ligularia stenocephala, and one medicinal plant, Panax ginseng, were cultivated in the understory of an artificial Larix kaempferi plantation for ten years. Growth characteristics (number of leaves and flower stalks per plant, and leaf length and width), survival rate, and yield (fresh weight of plants) per unit area (1 ㎡) were investigated one year after planting, and six and ten years following cultivation. P. ginseng and L. stenocephala survived at a high percentage for two years after planting. Results showed that P. ginseng had longer and thicker roots when aged; however, a large number of plants died and the yield was low. In contrast, almost half of A. victorialis var. platyphyllum died within two years of planting. The surviving plants grew well for ten years and the yield was increased. The leaf length and width of L. stenocephala increased; however, the survival rate and the number of leaves per plant decreased as the period of cultivation was extended. In contrast, A. victorialis var. platyphyllum survived at a lower rate (50%) than the two other crops (98% for L. stenocephala and 83% for P. ginseng) during the early cultivation period, with little change in the survival rate over an extended time; however, the yields increased. This species showed an increase in the number of flower stalks and leaves, and as a result, the larger leaves increased the yield. We evaluated the understory cultivation of three crops in a L. kaempferi plantation under natural conditions, with no irrigation or fertilization, and Allium victorialis var. platyphyllum showed the greatest growth potential among the three tested crops.

• Journal of Ginseng Research
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• v.36 no.1
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• pp.93-101
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• 2012

This study evaluated the anti-cholinesterases (ChEs) and antioxidant activities of white ginseng (WG) and black ginseng (BG) roots of Panax ginseng (PG), P. quinquefolium (PQ), and P. notoginseng (PN). Ginsenosides $Rg_1$, Re, Rf, $Rb_1$, Rc, $Rb_2$, and Rd were found in white PG, whereas Rf was not found in white PQ and Rf, Rc, and $Rb_2$ were not detected in white PN. The major ginsenoside content in steamed BG including $RK_3$, $Rh_4$, and 20(S)/(R)-$Rg_3$ was equivalent to approximately 70% of the total ginsenoside content. The WG and BG inhibited acetylcholinesteras (AChE) and butyrylcholinesterase (BChE) in a dose dependent manner. The efficacy of BG roots of PG, PQ, and PN on AChE and BChE inhibition was greater than that of the respective WG roots. The total phenolic contents and 2, 2-diphenyl-1-picryl-hydrazyl (DPPH) scavenging activity were increased by heat treatment. Among the three WG and BG, white PG and steamed black PQ have significantly higher contents of phenolic compounds. The best results for the DPPH scavenging activity were obtained with the WG and BG from PG. These results demonstrate that the steamed BG roots of the three studied ginseng species have both high ChEs inhibition capacity and antioxidant activity.

• Journal of Ginseng Research
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• v.44 no.1
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• pp.145-153
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• 2020

Background: Panax ginseng Meyer (Araliaceae) is a highly valued medicinal plant in Asian regions, especially in Korea, China, and Japan. Chemical and biological studies on P. ginseng have focused primarily on its roots, whereas the seeds remain poorly understood. This study explores the phytochemical and biological properties of compounds from P. ginseng seeds. Methods: P. ginseng seeds were extracted with methanol, and 16 compounds were isolated using various chromatographic methods. The chemical structures of the isolates were determined by spectroscopic data. Antiinflammatory activities were evaluated for triterpene and steroidal saponins using lipopolysaccharide-stimulated RAW264.7 macrophages and THP-1 monocyte leukemia cells. Results: Phytochemical investigation of P. ginseng seeds led to the isolation of a novel triterpene saponin, pseudoginsenoside RT₈, along with 15 known compounds. Pseudoginsenoside RT₈ exhibited more potent antiinflammatory activity than the other saponins, attenuating lipopolysaccharide-mediated induction of proinflammatory genes such as interleukin-1β, interleukin-6, inducible nitric oxide synthase, cyclooxygenase-2, and matrix metalloproteinase-9, and suppressed reactive oxygen species and nitric oxide generation in a dose-dependent manner. Conclusion: These findings indicate that pseudoginsenoside RT₈ has a pharmaceutical potential as an antiinflammatory agent and that P. ginseng seeds are a good natural source for discovering novel bioactive molecules.

• Journal of Ginseng Research
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• v.45 no.3
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• pp.442-449
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• 2021

Background: Panax ginseng is an important crop in Asian countries given its pharmaceutical uses. It is usually harvested after 4-6 years of cultivation. However, various abiotic stresses have led to its quality reduction. One of the stress causes is high content of heavy metal in ginseng cultivation area. Plant growth-promoting rhizobacteria (PGPR) can play a role in healthy growth of plants. It has been considered as a new trend for supporting the growth of many crops in heavy metal occupied areas, such as Aluminum (Al). Methods: In vitro screening of the plant growth promoting activities of five tested strains were detected. Surface-disinfected 2-year-old ginseng seedlings were dipping in Rhizobium panacihumi DCY116^T suspensions for 15 min and cultured in pots for investigating Al resistance of P. ginseng. The harvesting was carried out 10 days after Al treatment. We then examined H₂O₂, proline, total soluble sugar, and total phenolic contents. We also checked the expressions of related genes (PgCAT, PgAPX, and PgP5CS) of reactive oxygen species scavenging response and pyrroline-5-carboxylate synthetase by reverse transcription polymerase chain reaction (RT-PCR) method. Results: Among five tested strains isolated from ginseng-cultivated soil, R. panacihumi DCY116^T was chosen as the potential PGPR candidate for further study. Ginseng seedlings treated with R. panacihumi DCY116^T produced higher biomass, proline, total phenolic, total soluble sugar contents, and related gene expressions but decreased H₂O₂ level than nonbacterized Al-stressed seedlings. Conclusion: R. panacihumi DCY116^T can be used as potential PGPR and "plant strengthener" for future cultivation of ginseng or other crops/plants that are grown in regions with heavy metal exposure.

• Journal of Ginseng Research
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• v.46 no.4
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• pp.505-514
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• 2022

Background: The roots of Panax ginseng contain two types of tetracyclic triterpenoid saponins, namely, protopanaxadiol (PPD)-type saponins and protopanaxatiol (PPT)-type saponins. In P. ginseng, the protopanaxadiol 6-hydroxylase (PPT synthase) enzyme catalyses protopanaxatriol (PPT) production from protopanaxadiol (PPD). In this study, we constructed homozygous mutant lines of ginseng by CRISPR/Cas9-mediated mutagenesis of the PPT synthase gene and obtained the mutant ginseng root lines having complete depletion of the PPT-type ginsenosides. Methods: Two sgRNAs (single guide RNAs) were designed for target mutations in the exon sequences of the two PPT synthase genes (both PPTa and PPTg sequences) with the CRISPR/Cas9 system. Transgenic ginseng roots were generated through Agrobacterium-mediated transformation. The mutant lines were screened by ginsenoside analysis and DNA sequencing. Result: Ginsenoside analysis revealed the complete depletion of PPT-type ginsenosides in three putative mutant lines (Cr4, Cr7, and Cr14). The reduction of PPT-type ginsenosides in mutant lines led to increased accumulation of PPD-type ginsenosides. The gene editing in the selected mutant lines was confirmed by targeted deep sequencing. Conclusion: We have established the genome editing protocol by CRISPR/Cas9 system in P. ginseng and demonstrated the mutated roots producing only PPD-type ginsenosides by depleting PPT-type ginsenosides. Because the pharmacological activity of PPD-group ginsenosides is significantly different from that of PPT-group ginsenosides, the new type of ginseng mutant producing only PPD-group ginsenosides may have new pharmacological characteristics compared to wild-type ginseng. This is the first report to generate target-induced mutations for the modification of saponin biosynthesis in Panax species using CRISPR-Cas9 system.

• Journal of Ginseng Research
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• v.46 no.4
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• pp.536-542
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• 2022

Background: In aged skin, reactive oxygen species (ROS) induces degradation of the extracellular matrix (ECM), leading to visible aging signs. Collagens in the ECM are cleaved by matrix metalloproteinases (MMPs). Syringaresinol (SYR), isolated from Panax ginseng berry, has various physiological activities, including anti-inflammatory action. However, the anti-aging effects of SYR via antioxidant and autophagy regulation have not been elucidated. Methods: The preventive effect of SYR on skin aging was investigated in human HaCaT keratinocytes in the presence of H₂O₂, and the keratinocyte cells were treated with SYR (0-200 ㎍/mL). mRNA and protein levels of MMP-2 and -9 were determined by real-time PCR and Western blotting, respectively. Radical scavenging activity was researched by 2,2 diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) assays. LC3B level was assessed by Western blotting and confocal microscopy. Results: SYR significantly reduced gene expression and protein levels of MMP-9 and -2 in both H₂O₂-treated and untreated HaCaT cells. SYR did not show cytotoxicity to HaCaT cells. SYR exhibited DPPH and ABTS radical scavenging activities with an EC₅₀ value of 10.77 and 10.35 ㎍/mL, respectively. SYR elevated total levels of endogenous and exogenous LC3B in H₂O₂-stimulated HaCaT cells. 3-Methyladenine (3-MA), an autophagy inhibitor, counteracted the inhibitory effect of SYR on MMP-2 expression. Conclusion: SYR showed antioxidant activity and up-regulated autophagy activity in H₂O₂-stimulated HaCaT cells, lowering the expression of MMP-2 and MMP-9 associated with skin aging. Our results suggest that SYR has potential value as a cosmetic additive for prevention of skin aging.

• Journal of Veterinary Science
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• v.21 no.4
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• pp.61.1-61.16
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• 2020

Background: Panax notoginseng saponins (PNS) are bioactive substances extracted from P. notoginseng that are widely used to treat cardiovascular and cerebrovascular diseases and interstitial diseases. PNS have the functions of scavenging free radicals, anti-inflammation, improving blood supply for tissue and so on. Objectives: The aim of this study was to investigate the effects of PNS on the oxidative stress of immune cells induced by porcine circovirus 2 (PCV2) infection in vitro and in vivo. Methods: Using an oxidative stress model of PCV2 infection in a porcine lung cell line (3D4/2 cells) and mice, the levels of nitric oxide (NO), reactive oxygen species (ROS), total glutathione (T-GSH), reduced glutathione (GSH), and oxidized glutathione (GSSG) and the activities of xanthine oxidase (XOD), myeloperoxidase (MPO) and inducible nitric oxide synthetase (iNOS) were determined to evaluate the regulatory effects of PNS on oxidative stress. Results: PNS treatment significantly reduced the levels of NO and ROS, the content of GSSG and the activities of XOD, MPO, and iNOS (p < 0.05), while significantly increasing GSH and the ratio of GSH/GSSG in infected 3D4/2 cells (p < 0.05).Similarly, in the in vivo study, PNS treatment significantly decreased the level of ROS in spleen lymphocytes of infected mice (p < 0.05), increased the levels of GSH and T-GSH (p < 0.05), significantly decreased the GSSG level (p < 0.05), and decreased the activities of XOD, MPO, and iNOS. Conclusions: PNS could regulate the oxidative stress of immune cells induced by PCV2 infection in vitro and in vivo.