• Journal of Ginseng Research
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• v.47 no.2
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• pp.337-346
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• 2023

Background: Ginsenoside Rb2, a major active component of Panax ginseng, has various physiological activities, including anticancer and anti-inflammatory effects. However, the mechanisms underlying the rejuvenation effect of Rb2 in human skin cells have not been elucidated. Methods: We performed a senescence-associated β-galactosidase staining assay to confirm cellular senescence in human dermal fibroblasts (HDFs). The regulatory effects of Rb2 on autophagy were evaluated by analyzing the expression of autophagy marker proteins, such as microtubule-associated protein 1A/1B-light chain (LC) 3 and p62, using immunoblotting. Autophagosome and autolysosome formation was monitored using transmission electron microscopy. Autophagic flux was analyzed using tandem-labeled GFP-RFP-LC3, and lysosomal function was assessed with Lysotracker. We performed RNA sequencing to identify potential target genes related to HDF rejuvenation mediated by Rb2. To verify the functions of the target genes, we silenced them using shRNAs. Results: Rb2 decreased β-galactosidase activity and altered the expression of cell cycle regulatory proteins in senescent HDFs. Rb2 markedly induced the conversion of LC3-I to LC3-II and LC3 puncta. Moreover, Rb2 increased lysosomal function and red puncta in tandem-labeled GFP-RFP-LC3, which indicate that Rb2 promoted autophagic flux. RNA sequencing data showed that the expression of DNA damage-regulated autophagy modulator 2 (DRAM2) was induced by Rb2. In autophagy signaling, Rb2 activated the AMPK-ULK1 pathway and inactivated mTOR. DRAM2 knockdown inhibited autophagy and Rb2-restored cellular senescence. Conclusion: Rb2 reverses cellular senescence by activating autophagy via the AMPK-mTOR pathway and induction of DRAM2, suggesting that Rb2 might have potential value as an antiaging agent.

• Journal of Ginseng Research
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• v.37 no.3
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• pp.283-292
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• 2013

Ginsenosides are divided into two groups based on the types of the panaxadiol group (e.g., ginsenoside-Rb1 and -Rc) and the panaxatriol group (e.g., ginsenoside-Rg1 and -Re). Among them, ginsenoside-Re (G-Re) is one of the compounds with the highest content in Panax ginseng and is responsible for pharmacological effects. However, it is not yet well reported if G-Re increases the hemodynamics functions on ischemia (30 min)/reperfusion (120 min) (I/R) induction. Therefore, in the present study, we investigated whether treatment of G-Re facilitated the recovery of hemodynamic parameters (heart rate, perfusion pressure, aortic flow, coronary flow, and cardiac output) and left ventricular developed pressure (${\pm}dp/dt_{max}$). This research is designed to study the effects of G-Re by studying electrocardiographic changes such as QRS interval, QT interval and R-R interval, and inflammatory marker such as tissue necrosis factor-${\alpha}$ (TNF-${\alpha}$) in heart tissue in I/R-induced heart. From the results, I/R induction gave a significant increase in QRS interval, QT interval and R-R interval, but showed decrease in all hemodynamic parameters. I/R induction resulted in increased TNF-${\alpha}$ level. Treatment of G-Re at 30 and $100{\mu}M$ doses before I/R induction significantly prevented the decrease in hemodynamic parameters, ameliorated the electrocardiographic abnormality, and inhibited TNF-${\alpha}$ level. In this study, G-Re at $100{\mu}M$ dose exerted more beneficial effects on cardiac function and preservation of myocardium in I/R injury than $30{\mu}M$. Collectively, these results indicate that G-Re has distinct cardioprotectective effects in I/R induced rat heart.

• Korean Journal of Medicinal Crop Science
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• v.26 no.2
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• pp.148-156
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• 2018

Background: Hot steaming is known to be effective in improving the biological activities of plant extracts by breaking down useful compounds to low molecular weight ones. Methods and Results: This study aimed to develop an optimal extraction and steam processing method for enhancing the low molecular ginsenoside contents of the adventitious roots culture of wild mountain ginseng. The total ginsenoside was optimally extracted when 70% EtOH was used at $50^{\circ}C$, whereas low molecule ginsenoside such as Rg2, Rh1, Rh4 and Rk1 could be extracted using 70% EtOH at $70^{\circ}C$. The adventitious roots culture of wild mountain ginseng is known to contain four major ginsenosides, i.e., Rb2, Rb1, Rg1 and Rd, however new ginsenosides Rg6, Rh4, Rg3, Rk1 and Rg5 were new abundantly obtaind after steam processing method was applied. The contents of total ginsenosides were the highest when thermal steam processing was conducted at $120^{\circ}C$ for 120 min. Unlike ginsenosides such as Rg1, Re, Rb1, Rc, Rb2, and Rh1, which decreased after steam processing, Rg3, Rk1, and Rg5 increased after thermal processing. Steam processing significanltly reduced the content of Rb1, increased that of Rg6 by about ten times than that in the adventitious roots culture of wild mountain ginseng. Conclusions: Our study showed that the optimal extraction and steam processing method increased the content of total ginsenosides and allowed the extraction of minor ginsenosides from major ones.

• Journal of Ginseng Research
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• v.12 no.2
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• pp.104-113
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• 1988

Effect of the purified ginsenoside $-Rb_1$ and $-Rb_2$ on LDL receptor biosynthesis of CHO cell cultured in a high cholesterol medium was investigated . Cholesterol uptake by CHO cell cultured in a medium containing various amounts of cholesterol was traced and found that the cholesterol uptake was proportional to the concentration of cholesterol in the medium, and the population of LDL receptors were proportionally decreased as the increasing cholesterol level in the cell. However, when the CHO cells were cultured in the medium containing ginsenosides, no significant decrease of LDL receptor population occured. The biosynthesis of protein and RNA of the above cells was higher than that of CHO cells cultured in the absence of the ginsenosides, suggesting that the ginsenosides might stimulate LDL receptor bio-synthesis. It was also observed that the ginsenosides stimulated the biosynthesis of estradiol and progesterone from cholesterol in the CHO cell. From the above results, it seemed that the ginsenosides lowers the cholesterol level by stimulating the cholesterol metablism including steroid hormone biosynthesis, resulting in the lowering of inhibitory action of cholesterol on LDL receptor biosynthesis.

• Proceedings of the Ginseng society Conference
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• 1988.08a
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• pp.1-10
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• 1988

Previously. we reported that the intraperitoneal administration of ginseng crude saponin increased: (I) nuclear RNA polymerase activity. (2) nuclear RNA synthesis. (3) cytoplasmic RNA synthesis. (4) cytoplasmic heavy polyrioosome content. (5) amino acid incorporation in vitro of microsome and polysome isolated rat liver. and (6) the incorporation rate of labeled amino acids into serum protein. In addition, a spectacular increase in the rough endoplasmic reticulum of hepatocyte administered crude saponin for four weeks orally was shown through electron microscopy. An increase in polysomal content in membrane-hound ribosome was shown through ultracentrifugation. Recently, successive intraperitoneal. administration .of $ginsenosid-Rb_2$ was given to streptozotocin (STZ) diaoetic rats of hypoproteinemia. The blood urea nitrogen and hepatic urea concentration were decreased significantly. The total protein and alhumin levels in the serum were increased in comparison to control values. In contrast. the $ginsenoside-Rb_2$ treated group of STZ diahetic rats showed a significant increase in liver RNA. total ribosome and membrane-bound ribosomal contents. The administration of $ginsenoside-Rb_2$ increased the incorporation rate of labeled - precursor into total serum protein. Additionally $ginsenoside-Rb_2$ improved the nitrogen balance of diabetic rats. On the bases of these experimental results, ginseng saponin has a metabolic stimulatory or anabolic action on RNA and protein synthesis.

• Korean journal of food and cookery science
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• v.26 no.4
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• pp.475-480
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• 2010

In this study, the ginsenoside contents and anti-inflammatory effects of white ginseng (WG) and puffed red ginseng (PRG) were compared. The contents of Rb1, Rg5 and Rk1 were significantly higher in PRG than in WG, whereas the contents of Rg1 and Rb2 were decreased in PRG. The levels of NO production and iNOS expression were suppressed in LPS-stimulated cells by treatment with WG and PRG. Further, the production of cytokines (TNF-$\alpha$ and INF-$\gamma$) and inflammatory proteins (NF-${\kappa}B$ and COX-2) was decreased in cells upon treatment with any of the ginsenosides. The high NO inhibitory activity and cytokine production of PRG is caused by differences in the composition of ginsenosides produced.

• Proceedings of the Ginseng society Conference
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• 1988.08a
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• pp.47-54
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• 1988

Cholesterol a component of all eucaryotic plasma membranes. is essential for the growth and viability of cells in higher organisms. However. too much cholesterol can be lethal because of atherosclerosis resulting from the deposition of cholesterol ester plaques. It was attempted in this study to understand the preventive effect of ginseng saponin. one of the major components of the roots of Panax ginseng C.A. Meyer. against hypercholesterolemia induced by high cholesterol diet. $^{125}I-LDL$ was injected intravenously to rabbits and rats. which were fed a high cholesterol diet with and/or without ginseng saponin for 12 days. The disappearance of the radioactivity occurred faster in the test group than the control. The effect of saponin fraction from Panax ginseng C.A. Meyer and the purified ginsenosilks. $Rb_1,\;Rb_2,\;Re\;and\;Rg_1,$ on LDL receptor biosynthesis in high cholesterol fed rat has been investigated. Analysis of LDL receptors from various organs such as liver. kidney. adrenal cortex and testis showed that the population of LDL receptors of test group significantly higher than that of the control. It was also found that liver homogenate containing ginsenosides $(10^{-3}-10^{-4}\%)$ stimulated the biosynthesis of bile acid form cholesterol. From the above results. it seemed that ginsenosides lower the cholesterol level by stimulating cholesterol metabolism. which result in the suppression of the inhibitory action of cholesterol on LDL receptor biosynthesis.

• Proceedings of the Ginseng society Conference
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• 1980.09a
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• pp.59-69
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• 1980

A new HPLC-method for separation and quantitative determination of ginsenosides in Panax ginseng, Panax quinquefolium and in pharmaceutical drug preparations is elaborated. A reversed-phase-system with ${\mu}Bondapak\;C_{18}$ column (3.9 mm $I.D.{\times}30\;cm$) using acetonitrile-water (30:70) 2 ml/min and acetonitrile-water (18:82) 4 ml/min is suitable for the base-line separation of $Rb_1,\;Rb_2,\;Rc,\;Rd,\;Rf,\;Rg_2,\;respectively\;Re,\;Rg_1$ in 30 minutes. The ginsenosides are directly detected at 203 nm (without derivatization) with the LC-55 or LC-75 spectrophotometer (Perkin-Elmer) at $100\%$ transmission. Detection limit is 300 ng at a signal-to-noise ratio of 10:1. The ginsenosides-peak identification is carried out with HPTLC (high performance thin layer chromatography), with MIR-IR (multiple internal reflection-IR-spectros-copy) and with FD-MS (field desorption mass spectrometry). The calibration curve of each ginsenoside has a correlation coefficient very near to 1. Relative standard deviation for quantitative determinations depends upon the amount of ginsenosides and is approximately 1\%$ for ginsenoside contents of 1\%$. This method is adaptable for routine analysis in quality control laboratories.

• Proceedings of the PSK Conference
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• 2003.10b
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• pp.218.1-218.1
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• 2003

Saponins are known to be the major constituent of Panax ginseng. More than 30 kinds of ginseng saponins are reported so far. The major saponins in white ginseng (WG) or red ginseng (RG) are ginsenosides Rb1, Rb2, Rc, Rd, Rg1, and Re. HPLC method with ELSD or UV detection was used to analyze ginsenosides. Recently, a new processed ginseng with fortified activity, named as Sun Ginseng (SG), was reported. The major ginsenosides of SG are totally different from that of WG or RG, i.e., ginsenoside Rg3, Rk1, and Rg5 are the major constituents of SG. (omitted)

• Journal of Ginseng Research
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• v.21 no.3
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• pp.174-186
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• 1997

In this study, rat hepatocytes known to have active carbohydrate metabolism were obtained by using the liver perfusion technique to examine the hypoglycemic action of red ginseng saponin components [ginsenoside (mixture, $Rb_1$, and $Rg_1$)] and incubated in two different media-one containing insulin and glucagon (control group), and the other containing glucagon only, The specific activities of some regulatory enzymes such as glucokinase, glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, and glucose 6-phosphatase, in main pathways which were directly related to the glucose metabolism were compared between these two kinds of hepatocytes cultured in two different media. The effects of red ginseng saponin components [ginsenoside (mixture, $Rb_1$, and $Rg_1$)] under the concentration of $10^3$~$10^6$% on these enzymes In hepatocytes were also investigated, when they were added to these two media. The results were as follows. The specific activity of enzymes such as glucokinase, glucose 6-phosphate dehydrogenase, and 6-phosphogluconate dehydrogenase related to glucose-consuming pathways of insulin-deficient group was much less than control one, however, their decreased activity was recovered after the addition of ginseng components at all range of concentrations. The increased specific activity of these on - zymes was shown by the addition of ginseng components to the control group. On the other hand, the specific activity of glucose 6-phosphatase related to glucose-producing pathway of insulin-deficient group was much higher than control one, but their Increased activity was decreased after the addition of ginseng components at all range of concentrations. The same results were obtained after the addition of ginseng components to the control group. These results suggest that the red ginseng saponin components might better diabetic hyperglycemia by regulating the activity of enzymes related to glucose metabolism directly and/or Indirectly though more detailed studies were needed.