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A brief method for preparation of gintonin-enriched fraction from ginseng

  • Choi, Sun-Hye;Jung, Seok-Won;Kim, Hyun-Sook;Kim, Hyeon-Joong;Lee, Byung-Hwan;Kim, Joon Yong;Kim, Jung-Hyun;Hwang, Sung Hee;Rhim, Hyewon;Kim, Hyoung-Chun;Nah, Seung-Yeol
    • Journal of Ginseng Research
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    • v.39 no.4
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    • pp.398-405
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    • 2015
  • Background: Ginseng has been used as a tonic for invigoration of the human body. In a previous report, we identified a novel candidate responsible for the tonic role of ginseng, designated gintonin. Gintonin induces $[Ca^{2+}]_i$ transient in animal cells via lysophosphatidic acid receptor activation. Gintonin-mediated $[Ca^{2+}]_i$ transient is linked to anti-Alzheimer's activity in transgenic Alzheimer's disease animal model. The previous method for gintonin preparation included multiple steps. The aim of this study is to develop a simple method of gintonin fraction with a high yield. Methods: We developed a brief method to obtain gintonin using ethanol and water. We extracted ginseng with fermentation ethanol and fractionated the extract with water to obtain water-soluble and water-insoluble fractions. The water-insoluble precipitate, rather than the water-soluble supernatant, induced a large $[Ca^{2+}]_i$ transient in primary astrocytes. We designated this fraction as gintonin-enriched fraction (GEF). Results: The yield of GEF was approximately 6-fold higher than that obtained in the previous gintonin preparation method. The apparent molecular weight of GEF, determined using sodium dodecyl sulfate-polyacrylamide gel electrophoresis, was equivalent to that obtained in the previous gintonin preparation method. GEF induced $[Ca^{2+}]_i$ transient in cortical astrocytes. The effective dose (ED50) was $0.3{\pm}0.09{\mu}g/mL$. GEF used the same signal transduction pathway as gintonin during $[Ca^{2+}]_i$ transient induction in mouse cortical astrocytes. Conclusion: Because GEF can be prepared through water precipitation of ginseng ethanol extract and is easily reproducible with high yield, it could be commercially utilized for the development of gintoninderived functional health food and natural medicine.

A Simple Method for the Preparation of Crude Gintonin from Ginseng Root, Stem, and Leaf

  • Pyo, Mi-Kyung;Choi, Sun-Hye;Shin, Tae-Joon;Hwang, Sung-Hee;Lee, Byung-Hwan;Kang, Ji-Yeon;Kim, Hyeon-Joong;Lee, Soo-Han;Nah, Seung-Yeol
    • Journal of Ginseng Research
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    • v.35 no.2
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    • pp.209-218
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    • 2011
  • Ginseng has been used as a general tonic agent to invigorate the human body as an adaptogenic agent. In a previous report, we have shown that ginseng contains a novel glycolipoprotein called gintonin. The main function of gintonin is to transiently enhance intracellular free $Ca^{2+}$ $[Ca^{2+}]_i$ levels in animal cells. The previous method for gintonin isolation included multiple steps using organic solvents. In the present report, we developed a simple method for the preparation of crude gintonin from ginseng root as well as stem and leaf, which produced a higher yield of gintonin than the previous one. The yield of gintonin was 0.20%, 0.29%, and 0.81% from ginseng root, stem, and leaf, respectively. The apparent molecular weight of gintonin isolated from stem and leaf through sodium dodecyl sulfate polyacrylamide gel electrophoresis was almost same as that from root but the compositions of amino acids, carbohydrates or lipids differed slightly between them. We also examined the effects of crude gintonin from ginseng root, stem, and leaf on endogenous $Ca^{2+}$-activated $Cl^-$ channel (CaCC) activity of Xenopus oocytes through mobilization of $[Ca^{2+}]_i$. We found that the order of potency for the activation of CaCC was ginseng root > stem > leaf. The $ED_{50}$ was $1.4{\pm}1.4$, $4.5{\pm}5.9$, and $3.9{\pm}1.1$ mg/mL for root, stem and leaf, respectively. In the present study, we demonstrated for the first time that in addition to ginseng root, ginseng stem and leaf also contain gintonin. Gintonin can be prepared from a simple method with higher yield of gintonin from ginseng root, stem, and leaf. Finally, these results demonstrate the possibility that ginseng stem and leaf could also be utilized for ginstonin preparation after a simple procedure, rather than being discarded.

Effects of a gintonin-enriched fraction on hair growth: an in vitro and in vivo study

  • Lee, Na-Eun;Park, Sang-Deuk;Hwang, Hongik;Choi, Sun-Hye;Lee, Ra Mi;Nam, Sung Min;Choi, Jong Hee;Rhim, Hyewhon;Cho, Ik-Hyun;Kim, Hyoung-Chun;Hwang, Sung-Hee;Nah, Seung-Yeol
    • Journal of Ginseng Research
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    • v.44 no.1
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    • pp.168-177
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    • 2020
  • Background: Ginseng has been widely used as a health-promoting tonic. Gintonin present in ginseng acts as a lysophosphatidic acid (LPA) receptor ligand that activates six LPA receptor subtypes. The LPA6 subtype plays a key role in normal hair growth, and mutations in the LPA6 receptor impair normal human hair growth. Currently, human hair loss and alopecia are concerning issues that affect peoples' social and day-to-day lives. Objective: We investigated the in vitro and in vivo effects of a gintonin-enriched fraction (GEF) on mouse hair growth. Methods: Human hair follicle dermal papilla cells (HFDPCs) and six-week-old male C57BL/6 mice were used. The mice were divided into the four groups: control, 1% minoxidil, 0.75% GEF, and 1.5% GEF. The dorsal hair was removed to synchronize the telogen phase. Each group was treated topically, once a day, for 15 days. We analyzed hair growth activity and histological changes. Results: GEF induced transient [Ca2+]i, which stimulated HFDPC proliferation and caused 5-bromo-2'-deoxyuridine (BrdU) incorporation in a concentration-dependent manner. GEF-mediated HFDPC proliferation was blocked by the LPA receptor antagonist and Ca2+ chelator. HFDPC treatment with GEF stimulated vascular endothelial growth factor release. Topical application of GEF and minoxidil promoted hair growth in a dose-dependent manner. Histological analysis showed that GEF and minoxidil increased the number of hair follicles and hair weight. Conclusion: Topical application of GEF promotes mouse hair growth through HFDPC proliferation. GEF could be one of the main components of ginseng that promote hair growth and could be used to treat human alopecia.

The Inhibitory Effects of Korean Red Ginseng Saponins on 5- HT3A Receptor Channel Activity Are Coupled to Anti-Nausea and Anti-Vomiting Action

  • Kim Jong-Hoon;Lee Byung-Hwan;Jeong Sang Min;Nah Seung-Yeol
    • Journal of Ginseng Research
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    • v.29 no.1
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    • pp.37-43
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    • 2005
  • We performed in vitro and in vivo studies to know whether the inhibitory effects of ginsenosides on $5-HT_{3A}$ receptor channel acctivity are coupled to anti-nausea and anti-vomiting action. In vitro study, we investigated the effect of compound K (CK) and M4, which are ginsenoside metabolites, on human $5-HT_{3A}$ receptor channel activity expressed in Xenopus oocytes using two-electrode voltage clamp technique. Treatment of CK or M4 themselves had no effect in both oocytes injected with $H_2O\;and\;5-HT_{3A}$ receptor cRNA. In oocytes injected with $5- HT_{3A}$ receptor cRNA, M4 treatment inhibited more potently 5-HT-induced inward peak current $(I_{5-HT})$ than CK with dose-dependent and reversible manner. The half-inhibitory concentrations $(IC_{50})$ of CK and M4 were $36.9\;\pm\;10.1\;and\;7.3\;\pm\;2.2\;{\mu}M$, respectively. The inhibition of $I_{5-HT}$ by M4 was non-competitive and voltage-independent. These results indicate that M4 might regulate $5-HT_{3A}$ receptors. In vivo experiments, injection of cisplatin (7.5 mg/kg, i.v.) induced both nausea and vomiting with 1 h latency. These episodes reached to peak after 2 h and persisted for 4 h. Pre-treatment of GTS (500 mg/kg, p.o.) significantly reduced cisplatin-induced nausea and vomiting by $51\;\pm\;8.4\;and\;48.8\;\pm\;6.4\%$ during 4 h compared to GIS­untreated group, respectively. These results show the possibility that in vitro inhibition of $5-HT_{3A}$ receptor channel activity by ginsenosides might be coupled to in vivo anti-emetic activity.

The Antioxidant, Alcohol Metabolizing Enzyme, and Hepatoprotective Activities of Dendropanax morbifera Vinegar with Traditional Fermentation Methods (전통적인 발효 방법으로 제조한 황칠 식초의 항산화, 알코올 대사 효소 및 간보호 활성)

  • Jung, Kyung Im;Jung, Han Nah;Choi, Young Ju
    • Journal of Life Science
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    • v.32 no.4
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    • pp.290-297
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    • 2022
  • This study was performed to investigate the organic acids, alcohol metabolism enzyme, and antioxidative, nitrite-scavenging, and hepatoprotective effects of Dendropanax morbifera vinegar prepared by a traditional fermentation method. Among the organic acids detected, acetic acid was the highest found, at 91.72 mg/ml, followed by lactic acid (7.31 mg/ml), malic acid (1.36 mg/ml), and succinic acid (1.20 mg/ml). The total polyphenol content of the D. morbifera vinegar was 13.73 ㎍ tannic acid equivalent (TAE)/ml. The 1,1-Diphenyl-2-picrylhydrazy (DPPH) radical scavenging activity of D. morbifera vinegar was 76.04% at a 60% concentration. The superoxide dismutase (SOD) activity of D. morbifera vinegar was increased in a dose-dependent manner, which was 95.14% at a 60% concentration, while the α-glucosidase inhibitory activity of D. morbifera vinegar was 98.94% at a 10% concentration. The effects of D. morbifera vinegar on alcohol metabolism were determined by measuring the generation of reduced nicotinamide adenine dinucleotide (NADH) by alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH). The ADH and ALDH activities of D. morbifera vinegar were increased in a dose-dependent manner, 43.62% and 60.39% at a 60% concentration, respectively. The D. morbifera vinegar showed significant protective effects against tacrine-induced cytotoxicity in HepG2 cells at the 0.6% concentration. These results suggest that D. morbifera vinegar has great potential as a resource for high quality functional health beverages.

Oral Administration of Gintonin Attenuates Cholinergic Impairments by Scopolamine, Amyloid-β Protein, and Mouse Model of Alzheimer's Disease

  • Kim, Hyeon-Joong;Shin, Eun-Joo;Lee, Byung-Hwan;Choi, Sun-Hye;Jung, Seok-Won;Cho, Ik-Hyun;Hwang, Sung-Hee;Kim, Joon Yong;Han, Jung-Soo;Chung, ChiHye;Jang, Choon-Gon;Rhim, Hyewon;Kim, Hyoung-Chun;Nah, Seung-Yeol
    • Molecules and Cells
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    • v.38 no.9
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    • pp.796-805
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    • 2015
  • Gintonin is a novel ginseng-derived lysophosphatidic acid (LPA) receptor ligand. Oral administration of gintonin ameliorates learning and memory dysfunctions in Alzheimer's disease (AD) animal models. The brain cholinergic system plays a key role in cognitive functions. The brains of AD patients show a reduction in acetylcholine concentration caused by cholinergic system impairments. However, little is known about the role of LPA in the cholinergic system. In this study, we used gintonin to investigate the effect of LPA receptor activation on the cholinergic system in vitro and in vivo using wild-type and AD animal models. Gintonin induced $[Ca^{2+}]_i $ transient in cultured mouse hippocampal neural progenitor cells (NPCs). Gintonin-mediated $[Ca^{2+}]_i $ transients were linked to stimulation of acetylcholine release through LPA receptor activation. Oral administration of gintonin-enriched fraction (25, 50, or 100 mg/kg, 3 weeks) significantly attenuated scopolamine-induced memory impairment. Oral administration of gintonin (25 or 50 mg/kg, 1 2 weeks) also significantly attenuated amyloid-${\beta}$ protein ($A{\beta}$)-induced cholinergic dysfunctions, such as decreased acetylcholine concentration, decreased choline acetyltransferase (ChAT) activity and immunoreactivity, and increased acetylcholine esterase (AChE) activity. In a transgenic AD mouse model, long-term oral administration of gintonin (25 or 50 mg/kg, 3 months) also attenuated AD-related cholinergic impairments. In this study, we showed that activation of G protein-coupled LPA receptors by gintonin is coupled to the regulation of cholinergic functions. Furthermore, this study showed that gintonin could be a novel agent for the restoration of cholinergic system damages due to $A{\beta}$ and could be utilized for AD prevention or therapy.

Changes in the Usage of Dental Technology CAD/CAM (치과기공 CAD/CAM 사용에 대한 실태변화)

  • Nah, Jung-Sook
    • Journal of Technologic Dentistry
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    • v.37 no.4
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    • pp.271-284
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    • 2015
  • Purpose: The purpose of this study was to examine the usage of CAD/CAM, which is one of dental technologist duties, in an effort to look for any possible changes in the usage of CAD/CAM. It's specifically meant to compare the results of this study with those of Lee Jong-do, Park Kwang-Sig(2011)'s study in March, 2011, to determine if there were any changes over the past four years. The selected variables that were investigated were the ownership of CAD/CAM, awareness of it, the route of acquiring the first information on it, the merits and demerits of its usage, usage experience and educational experience about it. Methods: An online survey was conducted on the dental technologists who worked in urban communities including metropolitan cities to find out the usage of CAD/CAM in 2015 from July 15 to 31, 2015, after existing questionnaire items were modified. The collected data were analyzed by IBM SPSS statistics 22.0, and statistical data on frequency, percentage, mean and standard deviation for each item were obtained. To figure out the characteristics of the subjects and the relationship between their occupational characteristics and the usage of CAD/CAM, crosstabs, independent-samples t-test and one-way ANOVA were utilized. As for the usage of CAD/CAM in 2011, the results of Lee Jong-do, Park Kwang-Sig(2011)'s study were selected, and then a comparative analysis was made. The level of significance was all set at .05. Total 250 questionnaires were distributed to them, and 190(76.0%) were returned. After excluding 23 whose answers were uncertain or seemed to lack reliability, total 167(66.8%) were used in final analysis. Results: As a result of analyzing the usage of CAD/CAM that was one of dental technologist duties, there was a great increase in the ownership of CAD/CAM in their workplaces from 2.4 percent in 2011 to 71.7 percent in 2015, and there was an improvement in awareness about it and the necessity of its usage as well. In 2011, the Internet and mass media were the most common route that they got to know about it. They had a strong tendency to acquire related knowledge through education in 2015, and there were some changes in the products that they used or preferred. In both years, the great merit of CAD/CAM was the simplified manufacturing process of restoration, and it had another great advantages in 2015 such as the improved quality of restoration or improved environments for dental technology. Concerning disadvantages, high price was a big problem in 2011. In 2015, not only price but the burden of material costs, frequent breakdown, poor demand among dental clinics and a lack of CAD/CAM professionals were pointed out a lot. In the future, this researcher intends to make research to seek ways of improving CAD/CAM professionals. Conclusion: There was more awareness of CAD/CAM in 2015 than in 2011 when the changes in awareness of it were analyzed, and the finding suggest the necessity of sustained education and concern.

2-Layered Group Key Management Structure and Protocols using Multi-Core Based Tree (다중 코어 기반 트리를 이용한 2계층 그룹키 관리 구조 및 프로토콜)

  • Cho, Tac-Nam;Kim, Sang-Hee;Eun, Sang-A;Lee, Sang-Ho;Chae, Ki-Joon;Park, Won-Joo;Nah, Jae-Hoon
    • Journal of KIISE:Information Networking
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    • v.29 no.5
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    • pp.583-594
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    • 2002
  • Assuring the security of group communications such as tole-conference and software distribution requires a common group key be shared among the legal members in a secure manner. Especially for large groups with frequent membership change, efficient rekey mechanism is essential for scalability. One of the most popular ways to provide sealable rekey is to partition the group into several subgroups. In this paper, we propose a two-layered key management scheme which combines DEP and CBT, a protocol in which subgroup manager cannot access the multicast data and another that has a multi-core, respectively. We also select sub-group key management protocols suitable for our structure and design new rekey protocols to exclude the subgroup managers from the multicast data. Compared to previous protocols based on CBT, our scheme provides forward secrecy, backward secrecy and scalability. This would reduce the number of encryption and decryption for a rekey message and would improve the efficiency number of rekey messages and the amount of information related to group members that group managers must maintain compared to DEP.

Gintonin absorption in intestinal model systems

  • Lee, Byung-Hwan;Choi, Sun-Hye;Kim, Hyeon-Joong;Park, Sang-Deuk;Rhim, Hyewhon;Kim, Hyoung-Chun;Hwang, Sung-Hee;Nah, Seung-Yeol
    • Journal of Ginseng Research
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    • v.42 no.1
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    • pp.35-41
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    • 2018
  • Background: Recently, we identified a novel ginseng-derived lysophosphatidic acid receptor ligand, called gintonin. We showed that gintonin induces $[Ca^{2+}]i$ transient-mediated morphological changes, proliferation, and migration in cells expressing lysophosphatidic acid receptors and that oral administration of gintonin exhibits anti-Alzheimer disease effects in model mice. However, little is known about the intestinal absorption of gintonin. The aim of this study was to investigate gintonin absorption using two model systems. Methods: Gintonin membrane permeation was examined using a parallel artificial membrane permeation assay, and gintonin absorption was evaluated in a mouse everted intestinal sac model. Results: The parallel artificial membrane permeation assay showed that gintonin could permeate an artificial membrane in a dose-dependent manner. In the everted sac model, gintonin absorption increased with incubation time (from 0 min to 60 min), followed by a decrease in absorption. Gintonin absorption into everted sacs was also dose dependent, with a nonlinear correlation between gintonin absorption and concentration at 0.1-3 mg/mL and saturation at 3-5 mg/mL. Gintonin absorption was inhibited by the Rho kinase inhibitor Y-27632 and the sodiumeglucose transporter inhibitor phloridzin. Moreover, lipid extraction with methanol also attenuated gintonin absorption, suggesting the importance of the lipid portion of gintonin in absorption. This result shows that gintonin might be absorbed through passive diffusion, paracellular, and active transport pathways. Conclusion: The present study shows that gintonin could be absorbed in the intestine through transcellular and paracellular diffusion, and active transport. In addition, the lipid component of gintonin might play a key role in its intestinal absorption.

Differential Effects of Ginsenoside Metabolites on HERG K+ Channel Currents

  • Choi, Sun-Hye;Shin, Tae-Joon;Hwang, Sung-Hee;Lee, Byung-Hwan;Kang, Ji-Yeon;Kim, Hyeon-Joong;Oh, Jae-Wook;Bae, Chun-Sik;Lee, Soo-Han;Nah, Seung-Yeol
    • Journal of Ginseng Research
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    • v.35 no.2
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    • pp.191-199
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    • 2011
  • The human ether-a-go-go-related gene (HERG) cardiac $K^+$ channels are one of the representative pharmacological targets for development of drugs against cardiovascular diseases such as arrhythmia. Panax ginseng has been known to exhibit cardioprotective effects. In a previous report we demonstrated that ginsenoside $Rg_3$ regulates HERG $K^+$ channels by decelerating deactivation. However, little is known about how ginsenoside metabolites regulate HERG $K^+$ channel activity. In the present study, we examined the effects of ginsenoside metabolites such as compound K (CK), protopanaxadiol (PPD), and protopanaxatriol (PPT) on HERG $K^+$ channel activity by expressing human a subunits in Xenopus oocytes. CK induced a large persistent deactivatingtail current ($I_{deactivating-tail}$) and significantly decelerated deactivating current decay in a concentration-dependent manner. The $EC_{50}$ for persistent $I_{deactivating-tail}$ was $16.6{\pm}1.3$ ${\mu}M$. In contrast to CK, PPT accelerated deactivating-tail current deactivation. PPD itself had no effects on deactivating-tail currents, whereas PPD inhibited ginsenoside $Rg_3$-induced persistent $I_{deactivating-tail}$ and accelerated HERG $K^+$ channel deactivation in a concentration-dependent manner. These results indicate that ginsenoside metabolites exhibit differential regulation on Ideactivating-tail of HERG $K^+$ channel.