• Journal of Ginseng Research
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• v.43 no.2
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• pp.186-195
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• 2019

Background: Notoginseng stem-leaf (NGL) ginsenosides have not been well used. To improve their utilization, the biotransformation of NGL ginsenosides was studied using ginsenosidase type-I from Aspergillus niger g.848. Methods: NGL ginsenosides were reacted with a crude enzyme in the RAT-5D bioreactor, and the dynamic changes of multi-ginsenosides of NGL were recognized by HPLC. The reaction products were separated using a silica gel column and identified by HPLC and NMR. Results: All the NGL ginsenosides are protopanaxadiol-type ginsenosides; the main ginsenoside contents are 27.1% Rb3, 15.7% C-Mx1, 13.8% Rc, 11.1% Fc, 7.10% Fa, 6.44% C-Mc, 5.08% Rb2, and 4.31% Rb1. In the reaction of NGL ginsenosides with crude enzyme, the main reaction of Rb3 and C-Mx1 occurred through Rb3${\rightarrow}$C-Mx1${\rightarrow}$C-Mx; when reacted for 1 h, Rb3 decreased from 27.1% to 9.82 %, C-Mx1 increased from 15.5% to 32.3%, C-Mx was produced to 6.46%, finally into C-Mx and a small amount of C-K. When reacted for 1.5 h, all the Rb1, Rd, and Gyp17 were completely reacted, and the reaction intermediate F2 was produced to 8.25%, finally into C-K. The main reaction of Rc (13.8%) occurred through Rc${\rightarrow}$C-Mc1${\rightarrow}$C-Mc${\rightarrow}$C-K. The enzyme barely hydrolyzed the terminal xyloside on 3-O- or 20-O-sugar-moiety of the substrate; therefore, 9.43 g C-Mx, 6.85 g C-K, 4.50 g R7, and 4.71 g Fc (hardly separating from the substrate) were obtained from 50 g NGL ginsenosides by the crude enzyme reaction. Conclusion: Four monomer ginsenosides were successfully produced and separated from NGL ginsenosides by the enzyme reaction.

• Journal of Ginseng Research
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• v.40 no.1
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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.

• Mass Spectrometry Letters
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• v.7 no.4
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• pp.106-110
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• 2016

Ginseng, a traditional herbal drug, has been used in Eastern Asia for more than 2000 years. Various ginsenosides, which are the major bioactive components of ginseng products, have been shown to exert numerous beneficial effects on the human body when co-administered with drugs. However, this may give rise to ginsenoside-drug interactions, which is an important research consideration. In this study, acassette assay was performed the inhibitory effects of 12 ginsenosides on seven cytochrome P450 (CYP) isoforms in human liver microsomes (HLMs) using LC-MS/MS to predict the herb-drug interaction. After incubation of the 12 ginsenosides with seven cocktail CYP probes, the generated specific metabolites were quantified by LC-MS/MS to determine their activities. Ginsenoside Rb1 and F2 showed strong selective inhibitory effect on CYP2C9-catalyzed diclofenac 4'-hydroxylation and CYP2B6-catalyzed bupropion hydroxylation, respectively. Ginsenosides Rd showed weak inhibitory effect on the activities of CYP2B6, 2C9, 2C19, 2D6, 3A4, and compound K, while ginsenoside Rg3 showed weak inhibitory effects on CYP2B6. Other ginsenosides, Rc, Rf, Rg1, Rh1, Rf, and Re did not show significant inhibitory effects on the activities of the seven CYPs in HLM. Owing to the poor absorption of ginsenosides after oral administration in vivo, ginsenosides may not have significant side effects caused by interaction with other drugs.

• Journal of Ginseng Research
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• v.39 no.4
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• pp.392-397
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• 2015

Background: Red ginseng (RG) is processed from Panax ginseng via several methods including heat treatment, mild acid hydrolysis, and microbial conversion to transform the major ginsenosides into minor ginsenosides, which have greater pharmaceutical activities. During the fermentation process using microbial strains in a machine for making red ginseng, a change of composition occurs after heating. Therefore, we confirmed that fermentation had occurred using only microbial strains and evaluated the changes in the ginsenosides and their chemical composition. Methods: To confirm the fermentation by microbial strains, the fermented red ginseng was made with microbial strains (w-FRG) or without microbial strains (n-FRG), and the fermentation process was performed to tertiary fermentation. The changes in the ginsenoside composition of the self-manufactured FRG using the machine were evaluated using HPLC, and the 20 ginsenosides were analyzed. Additionally, we investigated changes of the reducing sugar and polyphenol contents during fermentation process. Results: In the fermentation process, ginsenosides Re, Rg1, and Rb1 decreased but ginsenosides Rh1, F2, Rg3, and Compound Y (C.Y) increased in primary FRG more than in the raw ginseng and RG. The content of phenolic compounds was high in FRG and the highest in the tertiary w-FRG. Moreover, the reducing sugar content was approximately three times higher in the tertiary w-FRG than in the other n-FRG. Conclusion: As the results indicate, we confirmed the changes in the ginsenoside content and the role of microbial strains in the fermentation process.

• Journal of Ginseng Research
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• v.42 no.2
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• pp.149-157
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• 2018

Background: Panax notoginseng leaves (PNL) exhibit extensive activities, but few analytical methods have been established to exclusively determine the dammarane triterpene saponins in PNL. Methods: Ultra-performance liquid chromatography coupled with time-of-flight mass spectrometry (UPLC/Q-TOF MS) and HPLC-UV methods were developed for the qualitative and quantitative analysis of ginsenosides in PNL, respectively. Results: Extraction conditions, including solvents and extraction methods, were optimized, which showed that ginsenosides Rc and Rb3, the main components of PNL, are transformed to notoginsenosides Fe and Fd, respectively, in the presence of water, by removing a glucose residue from position C-3 via possible enzymatic hydrolysis. A total of 57 saponins were identified in the methanolic extract of PNL by UPLC/Q-TOF MS. Among them, 19 components were unambiguously characterized by their reference substances. Additionally, seven saponins of PNL-ginsenosides Rb1, Rc, Rb2, and Rb3, and notoginsenosides Fc, Fe, and Fd-were quantified using the HPLC-UV method after extraction with methanol. The separation of analytes, particularly the separation of notoginsenoside Fc and ginsenoside Rc, was achieved on a Zorbax ODS C8 column at a temperature of $35^{\circ}C$. This developed HPLC-UV method provides an adequate linearity ($r^2$ > 0.999), repeatability (relative standard deviation, RSD < 2.98%), and inter- and intraday variations (RSD < 4.40%) with recovery (98.7-106.1%) of seven saponins concerned. This validated method was also conducted to determine seven components in 10 batches of PNL. Conclusion: These findings are beneficial to the quality control of PNL and its relevant products.

• Journal of Ginseng Research
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• v.14 no.2
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• pp.178-186
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• 1990

The analgesic effect of morphine was antagonized and the development of tolerance was suppressed by the modification of the neurologic function in the animals treated with ginseng saponins The activation of the spinal descending inhibitory systems as well as the supraspinal structures by the administration of morphine was inhibited in the animals treated with ginseng saponine intracerebrally or intrathecally. The development of morphine tolerance and dependence, and the abrupt expression of naloxone inducted abstinence syndrom were also inhibited by ginsenoside Rb1, Rb2, Rg1 and Re. These results suggest that ginsenoside Rbl, Hbs, Rgl and Re are the bioactive components of panax ginseng on the inhibition of the development of morphine tolerance and dependence, and the inhibition of abrupt abstinence sindrome. In addition, further research on the minor components of Pnnnxkinsenl should be investigated. A single or daily treatment with ginseng saponins did not induce any appreciable changes in the brain in level of monoamines at the variolls time intervals and at the various day intervals, respectively. The inhibitory or facilitated effects of ginseng saponins on electrically evoked contractions in guinea pig ileum ($\mu$-receptor) and mouse vats deferens ($\delta$-receptor) were not mediated through opioid receptors. The antagonism of a $\chi$ receptor agonist, U-50, 488H was also not mediated through opioid receptors in the animals treated with ginseng saponins, but mediated through serotonergic mechanisms. Ginseng saponins inhibited morphine 6-dehydrogenase which catalyzed the production of morphinone from morphine, and increased hepatic glutathione contents for the detoxication of morphinone. This result suggests that the dual action of the above plays an important role in the inhibition of the development of morphine tolerance and dependence.

• Journal of Ginseng Research
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• v.43 no.2
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• pp.172-178
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• 2019

Inflammation is an innate immune response that protects the body from pathogens, toxins, and other dangers and is initiated by recognizing pathogen-associated molecular patterns or danger-associated molecular patterns by pattern-recognition receptors expressing on or in immune cells. Intracellular pattern-recognition receptors, including nucleotide-binding oligomerization domain-like receptors (NLRs), absent in melanoma 2, and cysteine aspartate-specific protease (caspase)-4/5/11 recognize various pathogen-associated molecular patterns and danger-associated molecular patterns and assemble protein complexes called "inflammasomes." These complexes induce inflammatory responses by activating a downstream effector, caspase-1, leading to gasdermin D-mediated pyroptosis and the secretion of proinflammatory cytokines, such as interleukin $(IL)-1{\beta}$ and IL-18. Ginsenosides are natural steroid glycosides and triterpene saponins found exclusively in the plant genus Panax. Various ginsenosides have been identified, and their abilities to regulate inflammatory responses have been evaluated. These studies have suggested a link between ginsenosides and inflammasome activation in inflammatory responses. Some types of ginsenosides, including Rh1, Rg3, Rb1, compound K, chikusetsu saponin IVa, Rg5, and Rg1, have been clearly demonstrated to inhibit inflammatory responses by suppressing the activation of various inflammasomes, including the NLRP3, NLRP1, and absent in melanoma 2 inflammasomes. Ginsenosides have also been shown to inhibit caspase-1 and to decrease the expression of $IL-1{\beta}$ and IL-18. Given this body of evidence, the functional relationship between ginsenosides and inflammasome activation provides new insight into the understanding of the molecular mechanisms of ginsenoside-mediated antiinflammatory actions. This relationship also has applications regarding the development of antiinflammatory remedies by ginsenoside-mediated targeting of inflammasomes, which could be used to prevent and treat inflammatory diseases.

• Korean Journal of Medicinal Crop Science
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• v.28 no.1
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• pp.9-20
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• 2020

Background: Although a number of Panax ginseng cultivars have been developed by Korean researchers in recent years, there has been insufficient analysis of their beneficial properties. In this study, we sought to identify useful ginseng varieties as functional materials. Methods and Results: We evaluated effects of root extracts of 10 ginseng cultivars (Cheongsun; CS, Chunpoong; CP, Gopoong; GP, Gumpoong; GMP, K1, Sunhyang; SH, Sunone; SO, Sunpoong; SP, Sunun; SU and Yunpoong; YP) against the inhibitory effects of nitric oxide (NO) and reactive oxygen species (ROS) production in mouse brain microglial BV2 cells, as well as the binding of N-methyl-D-aspartate receptor (NMDAR), a marker related to memory. Ginsenosides, such as 20 (S)-protopanaxadiols (PPDs), including ginsenoside-Rb1, -Rb2, -Rb3, -Rc, -Rd, and - Rg3 and 20 (S)-protopanaxatriols (PPTs) including -Re, -Rg1, and -Rg2 were analyzed by HPLC. We observed that the cultivar GMP showed the highest inhibitory effect (60.8%) against NO production at 20 ㎍/㎖. Those cultivars showing the significantly highest inhibition effects against ROS at 20 ㎍/㎖ were K1 (57.3%), SP (54.5%), YP (53.1%), CP (51.7%), CS (50.9%) and SH (49.6%). At 50 ㎍/㎖, K1 showed the most potent inhibitory effect (51.2%) on NMDAR binding. The total phenol content of SH (1.89 mg/g) and K1 (1.73 mg/g) were higher than those of the other cultivars, whereas in terms of PD/PT ratios, the values of CP (0.98), K1 (1.05) and SO (1.05) were lower than those of the other cultivars. On the basis of correlation coefficient (0.7064) between NMDAR inhibition and ONOO^- scavenging activity. Conclusions: The findings of this study indicate that the cultivars K1 and SH could be useful ginseng resources as functional materials with favorable cognition-improving and antioxidative properties.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.37 no.1
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• pp.75-81
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• 2011

The effect of lactic acid bacteria.fermented ginseng berry extract (FGBE) on physiological activities was evaluated. The contents of ginsenosides Re, Rc, and Rb1 in ginseng berry extract (GBE) were increased after fermentation by lactic acid bacteria when analyzed by high performance liquid chromatography. Antioxidant activity of GBE and FGBE was also analyzed by DPPH radical scavenging activity assay and SOD.like activity assay. FGBE showed a 86.34 % inhibition of DPPH radical and a 76.82 % inhibition by SOD.like activity at a concentration of 1.00 %. GBE showed a 49.78 % inhibition of DPPH radical and a 40.80 % inhibition by SOD.like activity at the same concentration. Furthermore, procollagen type I (COL1A1) gene expression increased by 823.13 % and matrix metal-loproteinase(MMP)-1 and tumor necrosis factor (TNF)-${\alpha}$ gene expression decreased by 87.88 % and 99.92 %, respectively, in human fibroblast cultured with FGBE at a concentration of 0.50 %. These results suggest that FGBE could be used as an active ingredient for functional cosmetics.

• Korean Journal of Pharmacognosy
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• v.46 no.4
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• pp.352-364
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• 2015

The aim of this study was to quantitatively analyze for quality assessment of eighteen marker compounds, including homogentisic acid, 3,4-dihydroxybenzaldehyde, spinosin, liquiritin, hesperidin, ginsenoside Rg1, liquiritigenin, ginsenoside Rb1, glycyrrhizin, 6-gingerol, atractylenolide III, honokiol, costunolide, dehydrocostuslactone, atractylenolide II, nootkatone, magnolol, and atractylenolide I, in Hyangsayukgunja-tang using an ultra-performance liquid chromatography-electrospray ionization-mass spectrometer. The column for separation of eighteen marker components were used a UPLC BEH $C_{18}$ analytical column ($2.1{\times}100mm$, $1.7{\mu}$) and kept at $45^{\circ}C$ by gradient elution with 0.1% (v/v) formic acid in water and acetonitrile as mobile phase. The flow rate and injection volume were 0.3 mL/min and $2.0{\mu}l$, respectively. The correlation coefficient of all marker compounds was ${\geq}0.9914$, which means good linearity, within the test ranges. The limits of detection and quantification values of the all analytes were in the ranges 0.04-1.11 and 0.13-3.33 ng/mL, respectively. As a result, five compounds, homogentisic acid, 3,4-dihydroxybenzaldehyde, spinosin, liquiritigenin, and atractylenolide I, in this sample were not detected and the amounts of the 13 compounds except for the 5 compounds were $8.10-6736.37{\mu}g/g$ in Hyangsayukgunja-tang extract.