• Korean Journal of Medicinal Crop Science
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• v.22 no.3
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• pp.223-230
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• 2014

This study was performed to enhance contents of low molecular ginsenoside using steaming and fermentation process in low quality fresh ginseng. For increase in contents of Rg2, Rg3, Rh2 and CK in low quality fresh ginseng, a steaming process was applied at $90^{\circ}C$ for 12 hr which was followed by fermentation process at Lactobacillus rhamnosus HK-9 incubated at $36^{\circ}C$ for 72 h. The contents of ginsenoside Rg1, Rb1, Rc, Re and Rd were decreased with the steaming associated with fermentation process but ginsenoside Rg2, Rg3, Rh2 and CK increased after process. It was found that under the steaming associated with fermentation process, low molecule ginsenosides such as Rg2, Rg3, Rh2 and CK were increased as 3.231 mg/g, 2.585 mg/g and 1.955 m/g and 2.478 mg/g, respectively. In addition, concentration of benzo[${\alpha}$]pyrene in extracts of the low quality fresh ginseng treated by the complex process was 0.11 ppm but it was 0.22 ppm when it was treated with the steaming process. This result could be caused by that the most efficiently breakdown of 1,2-glucoside and 1,4-glucoside linkage to backbone of ginsenosides by steaming associated with fermentation process. This results indicate that steaming process and fermenration process can increase in contents of Rg2, Rg3, Rh2 and CK in low quality fresh ginseng.

• Korean Journal of Pharmacognosy
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• v.45 no.1
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• pp.77-83
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• 2014

The purpose of this study is to develop a new preparation process of ginseng extracts having high concentrations of ginsenoside $Rg_3$, $Rg_5$ and $Rk_1$, a special component of Red ginseng. Chemical transformation from ginseng saponin glycosides to prosapogenin was analyzed by the HPLC. Extracts of White ginseng (Panax ginseng) and Fine White ginseng were processed under several treatment conditions including microwave and vinegar (about 14% acidity) treatments. Results of those treatments showed that the quantity of ginsenoside $Rg_3$ increased by over 0.6% at 4 minutes of pH 2~4 vinegar and microwave treatments. The results of processing with MWG-4 indicate that the Microwave and vinegar processed white ginseng extracts (about 14% acidity) that had gone through 4-minute treatments were found to contain the largest amount of ginsenoside $Rg_3$ (0.626%), $Rg_5$ (0.514%) and $Rk_1$ (0.220%). Results of treatments with MFWG-5 showed that the Fine White ginseng extracts that had been processed with microwave and vinegar (about 14% acidity) for 5 minutes were found to contain the largest amount of ginsenoside $Rg_3$ (4.484%), $Rg_5$ (3.192%) and $Rk_1$ (1.684%). It is thought that such results provide basic information in preparing White ginseng and Fine White ginseng extracts with functionality enhanced.

• Journal of Ginseng Research
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• v.41 no.1
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• pp.78-84
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• 2017

Background: In the present study, metabolite profiles of ginsenosides Rk1 and Rg5 from red ginseng or red notoginseng in zebrafish were qualitatively analyzed with ultraperformance liquid chromatography/quadrupole-time-of-flight MS, and the possible metabolic were pathways proposed. Methods: After exposing to zebrafish for 24 h, we determined the metabolites of ginsenosides Rk1 and Rg5. The chromatography was accomplished on UPLC BEH C18 column using a binary gradient elution of 0.1% formic acetonitrile-0.1% formic acid water. The quasimolecular ions of compounds were analyzed in the negative mode. With reference to quasimolecular ions and MS2 spectra, by comparing with reference standards and matching the empirical molecular formula with that of known published compounds, and then the potential structures of metabolites of ginsenosides Rk1 and Rg5 were acquired. Results: Four and seven metabolites of ginsenoside Rk1 and ginsenoside Rg5, respectively, were identified in zebrafish. The mechanisms involved were further deduced to be desugarization, glucuronidation, sulfation, and dehydroxymethylation pathways. Dehydroxylation and loss of C-17 residue were also metabolic pathways of ginsenoside Rg5 in zebrafish. Conclusion: Loss of glucose at position C-3 and glucuronidation at position C-12 in zebrafish were regarded as the primary physiological processes of ginsenosides Rk1 and Rg5.

• Korean Journal of Microbiology
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• v.54 no.4
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• pp.436-441
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• 2018

Ginseng are native traditional herbs, which exhibit excellent pharmacological activities. Probiotic Lactobacillus helveticus KII13 and Pediococcus pentosaceus strain KID7 were used for ginsenoside transformation by fermenting crude ginseng extract to enhance minor gisenoside content. Thin-layer chromatography (TLC) analysis of fermented ginseng extract showed that the minor ginsenosides Rg3, Rh1, and Rh2 were main products after 5 days of fermentation. HPLC analysis was performed to quantify the major and minor ginsenosides. The Rg3 peak appeared on the 3rd day while the appearance of Rh2 peak and Rh1 peak were observed on the 5th day. The co-culture of L. helveticus KII13 and P. pentosaceus KID7 converted major ginsenosides (Rb1 and Rg1) into minor ginsenosides (Rg3, Rh2, and Rh1).

• Animal Bioscience
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• v.35 no.7
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• pp.989-998
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• 2022

Objective: This study investigated the effects of intrauterine growth restriction (IUGR) during late pregnancy on the cell growth, proliferation, and differentiation in ovine fetal thymuses. Methods: Eighteen time-mated Mongolian ewes with singleton fetuses were allocated to three groups at d 90 of pregnancy: restricted group 1 (RG1, 0.18 MJ ME/body weight [BW]^0.75/d, n = 6), restricted group 2 (RG2, 0.33 MJ ME/BW^0.75/d, n = 6) and control group (CG, ad libitum, 0.67 MJ ME/BW^0.75/d, n = 6). Fetuses were recovered at slaughter on d 140. Results: The G0/G1 phase cell number in fetal thymus of the RG1 group was increased but the proliferation index and the expression of proliferating cell nuclear antigen (PCNA) were reduced compared with the CG group (p<0.05). Fetuses in the RG1 group exhibited decreased growth hormone receptor (GHR), insulin-like growth factor 2 receptor (IGF-2R), and their mRNA expressions (p<0.05). For the RG2 fetuses, there were no differences in the proliferation index and PCNA expression (p>0.05), but growth hormone (GH) and the mRNA expression of GHR were lower than those of the CG group (p<0.05). The thymic mRNA expressions of cyclin-dependent protein kinases (CDKs including CDK1, CDK2, and CDK4), CCNE, E2-factors (E2F1, E2F2, and E2F5) were reduced in the RG1 and RG2 groups (p<0.05), and decreased mRNA expressions of E2F4, CCNA, CCNB, and CCND were occurred in the RG1 fetuses (p<0.05). The decreased E-cadherin (E-cad) as a marker for epithelial-mesenchymal transition (EMT) was found in the RG1 and RG2 groups (p<0.05), but the OB-cadherin which is a marker for activated fibroblasts was increased in fetal thymus of the RG1 group (p<0.05). Conclusion: These results indicate that weakened GH/IGF signaling system repressed the cell cycle progression in G0/G1 phase in IUGR fetal thymus, but the switch from reduced E-cad to increased OB-cadherin suggests that transdifferentiation process of EMT associated with fibrogenesis was strengthened. The impaired cell growth, retarded proliferation and modified differentiation were responsible for impaired maturation of IUGR fetal thymus.

• Journal of Microbiology and Biotechnology
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• v.27 no.3
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• pp.616-623
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• 2017

Ginsenoside Rg3, a saponin extracted from ginseng, has various pharmacological and biological activities; however, its effects against Brucella infection are still unclear. Herein, the inhibitory effects of ginsenoside Rg3 against intracellular parasitic Brucella infection were evaluated through bacterial infection, adherence assays, and LAMP-1 colocalization, as well as immunoblotting and FACS for detecting MAPK signaling proteins and F-actin polymerization, respectively. The internalization, intracellular growth, and adherence of Brucella abortus in Rg3-treated RAW 264.7 cells were significantly decreased compared with the Rg3-untreated control. Furthermore, an apparent reduction of F-actin content and intensity of F-actin fluorescence in Rg3-treated cells was observed compared with B. abortus-infected cells without treatment by flow cytometry analysis and confocal microscopy, respectively. In addition, treating cells with Rg3 decreased the phosphorylation of MAPK signaling proteins such as ERK 1/2 and p38 compared with untreated cells. Moreover, the colocalization of B. abortus-containing phagosomes with LAMP-1 was markedly increased in Rg3-treated cells. These findings suggest that ginsenoside Rg3 inhibits B. abortus infection in mammalian cells and can be used as an alternative approach in the treatment of brucellosis.

• Journal of Ginseng Research
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• v.22 no.2
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• pp.133-139
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• 1998

We have studied an activation mechanism of $pp60^{c-src}$ protein tyroslne kinase (PTK) by ginsenoside-$Rg_1$ (G-$Rg_1$ ) in NIH(pMcsrc/foc)B c-src overexpressor cells. It was previously reported that G--$Rg_1$ stimulated the activation of c-src kinase at 20 pM with a 18 hr-incubation, increasing the activity by 2-4-fold over that of untreated control, and this effect was blocked by treatments of in- hibitors of either protein synthesis (cycloheximide) or RNA synthesis (actinomycin D) (Hong, H.Y. et at. Arch. Pharm. Res. 16, 114 (1993)). However, an amount of c-src protein itself in wild-type cells was not changed by G-$Rg_1$. When the cells mutated at one or two tyrosine residue(s) (Y416/527) that are important sites to regulate the kinase activity were treated with G-$Rg_1$, increases both in the activity of c-src kinase and in the expression of the protein were not observed. In addition, removal of extracellular calcium ion by EGTA or inhibition of PKC by H-7 canceled the G-$Rg_1$-induced activation of the kinase. Although the activation was little affected by G-$Rg_1$ with a calcium ionophore A23187, it was synergistically stimulated by treatment of G-Rgl and PMA, a PKC activator. Taken together, these results suggest that the activation of c-src kinase by G-$Rg_1$ is caused by an increase in the specific activity of the kinase, but not in amount of it, and is involved with both collular calcium ion and PKC. Further the increase in the specific activity of c-src kinase may result from altered phosphorylation at tyro-416 and -527.

• Journal of Microbiology and Biotechnology
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• v.33 no.4
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• pp.441-448
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• 2023

Brucellosis is a contagious zoonotic disease that infects millions of people annually with hundreds of millions more being exposed. It is caused by Brucella, a highly infectious bacterial species capable of infecting humans with an estimated dose of 10-100 organisms. Sirtuin 1 (SIRT1) has been reported to contribute to prevention of viral diseases as well as a chronic infection caused by Mycobacterium bovis. Here, we investigated the role of SIRT1 in the establishment of Brucella abortus infection in both in vitro and in vivo systems using the reported SIRT1 activators resveratrol (RES), piceatannol (PIC), and ginsenoside Rg3 (Rg3). In RAW264.7 cells, SIRT1 activators did not alter the adherence of Brucella or Salmonella Typhimurium. However, reduced uptake of Brucella was observed in cells treated with PIC and Rg3, and survival of Brucella within the cells was only observed to decrease in cells that were treated with Rg3, while PIC treatment reduced the intracellular survival of Salmonella. SIRT1 treatment in mice via oral route resulted in augmented Brucella resistance for PIC and Rg3, but not RES. PIC treatment favors Th2 immune response despite reduced serum pro-inflammatory cytokine production, while Rg3-treated mice displayed high IL-12 and IFN-γ serum production. Overall, our findings encourage further investigation into the complete mechanisms of action of the different SIRT1 activators used as well as their potential benefit as an effective alternative approach against intracellular and extracellular pathogens.

• Molecular & Cellular Toxicology
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• v.5 no.2
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• pp.126-132
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• 2009

The usage and types of chemicals are advancing, specializing, large-scaled increasing, and new chemical exposed workers are concerning to occupational disease. The generation of reactive oxygen in the body from carcinogen, mutation and DNA damage in cancer is protected by natural antioxidants (phytochemicals) with antimutagenic effect. There were many reports of ginsenoside Rb$_1$, Rg$_1$ grievances of the genetic mutation to suppress the effect confirm the genetic toxicity test with chromosomal aberration test and the Comet (SCGE) assay confirmed the suppression effect occurring chromosomal DNA damage. We had wanted to evaluate the compatibility and sensitivity between the chromosomal aberration (CA) test and the Comet assay. We used the CA test and Comet assay to evaluate the anti-genotoxicity of ginsenoside Rb$_1$ and Rg$_1$, in CHO-K1 (Chinese hamster ovary fibroblast) cell in vitro, composed negative control (solvent), positive control (benzo[a]pyrene), test group (carcinogen+variety concentration of ginsenoside) group. The positive control was benzo[a]pyrene (50 $\mu$M), well-known carcinogen, and the negative control was the 1 % DMSO solvent. The test group was a variety concentration of ginsenoside Rb$_1$, Rg$_1$ with 10$^{-8}$%, 10$^{-6}$%, 10$^{-4}$%, 10$^{-2}$%, 1%, 10%. In chromo-somal aberration test, we measured the number of cells with abnormally structured chromosome. In Comet assay, the Olive tail moment (OTM) and Tail length (TL) values were measured. The ratio of cell proliferation was increased 8.3% in 10$^{-8}$%, 10$^{-6}$%, 10$^{-4}$%, 10$^{-2}$%, 1%, 10% Rb$_1$ treated groups, and increased 10.4% in 10$^{-10}$%, 10$^{-8}$%, 10$^{-6}$%, 10$^{-4}$%, 10$^{-2}$%, 1% Rg$_1$ treated groups. In the CA test, the number of chromosomal aberration was decreased all the Rb$_1$ and Rg$_1$ treated groups. In the Comet assay, the OTM values were decreased in all the Rb$_1$ and Rg$_1$ treated groups. To evaluate the compatibility between CA and Comet assay, we compared the reducing ratio of chromosomal abnormalities with its OTM values, it was identified the antimutagenicity of ginsenoside, but it was more sensitive the CA test than the Comet assay. Ginsenoside Rb$_1$ and Rg$_1$ significantly decrease the number of cells with chromosomal aberration, and decrease the extent of DNA migration. Therefore, ginsenoside Rb$_1$, Rg$_1$ are thought as an antioxidant phytochemicals to protect mutagenicity. The in vitro Comet assay seems to be less sensitive than the in vitro chromosomal aberration test.

• Analytical Science and Technology
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• v.11 no.5
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• pp.404-408
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• 1998

Using a reversed-phase high performance liquid chromatography, the separation of 20(S)-, 20(R)-prosapogenin stereo-isomers of ginsenoside-$Rg_2$ and of ginsenoside-$Rg_3$ in ginseng saponins has been carried out with binary solvent system. The optimum conditions for the isomer separation are as following: Nova-$Pak^{(R)}C_{18}$ (Waters, $3.9{\times}150mm$) column, $CH_3CN/CH_3CN$ (100:8, v/v) binary solvent system and the flow rate was 1.7 mL/min. The stereoisomers were separated with change of the mixture ratio of the solvent system, the solvent elution by gradient program, and then detected at 203 nm of UV detector. The simultaneous separation of mixture that were the $Rg_2$, $Rg_3$ isomers was easily performed in nonpolar solvent for $Rg_2$, polar solvent for $Rg_3$ at the same optimum conditions.