• 한국약용작물학회지
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• 제26권2호
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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.

• 한국약용작물학회지
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• 제20권2호
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• pp.117-123
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• 2012

This study compared the contents of low molecular ginsenoside according to fermentation process in low grade fresh ginseng. Low grade fresh ginseng was directly inoculated with a 24 h seed culture of $Bifidobacterium$ Longum B6., $Lactobacillus$ $casei$., and incubated at $36^{\circ}C$ for 72 h. $Bifidobacterium$ Longum B6 was specifically was found to show the best growth on $3,255{\times}10^6\;CFU/m{\ell}$ after 48 h of fermentation. The content of ginsenoside Rb1, Re and Rd were decreased with the fermentation but ginsenoside Rh2 and Rg2 increased after fermentation process. In the case of low molecular ginsenoside conversion yields were 56.07% of Rh2, 12.03% of Rg3 and 77.11% of Rg2, respectively. In addition, compound-K was irregular conversion yield as long as 72 h of fermentation. This results indicate that fermentation process could increase the low molecular ginsenoside in low grade fresh ginseng.

• Journal of Ginseng Research
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• 제40권2호
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• pp.105-112
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• 2016

Background: Minor saponins or human intestinal bacterial metabolites, such as ginsenosides Rg3, F2, Rh2, and compound K, are more pharmacologically active than major saponins, such as ginsenosides Rb1, Rb2, and Rc. In this work, enzymatic hydrolysis of ginsenoside Rb1 was studied using enzyme preparations from cultured mycelia of mushrooms. Methods: Mycelia of Armillaria mellea, Ganoderma lucidum, Phellinus linteus, Elfvingia applanata, and Pleurotus ostreatus were cultivated in liquid media at $25^{\circ}C$ for 2 wk. Enzyme preparations from cultured mycelia of five mushrooms were obtained by mycelia separation from cultured broth, enzyme extraction, ammonium sulfate (30-80%) precipitation, dialysis, and freeze drying, respectively. The enzyme preparations were used for enzymatic hydrolysis of ginsenoside Rb1. Results: Among the mushrooms used in this study, the enzyme preparation from cultured mycelia of A. mellea (AMMEP) was found to convert ginsenoside Rb1 into compound K with a high yield, while those from G. lucidum, P. linteus, E. applanata, and P. ostreatus produced remarkable amounts of ginsenoside Rd from ginsenoside Rb1. The enzymatic hydrolysis pathway of ginsenoside Rb1 by AMMEP was $Rb1{\rightarrow}Rd{\rightarrow}F2{\rightarrow}$ compound K. The optimum reaction conditions for compound K formation from ginsenoside Rb1 were as follows: reaction time 72-96 h, pH 4.0-4.5, and temperature $45-55^{\circ}C$. Conclusion: AMMEP can be used to produce the human intestinal bacterial metabolite, compound K, from ginsenoside Rb1 with a high yield and without food safety issues.

• 생약학회지
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• 제47권4호
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• pp.352-359
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• 2016

The purpose of this study is to develop a new preparation process of ginseng leaf and stem extracts having high concentrations of ginsenoside Rg2, Rg3, Rg5, Rh1, a special component of red and black ginseng. Chemical transformation from ginseng saponin glycosides to prosapogenin was analyzed by the HPLC. Extracts of ginseng (Panax ginseng) leaf and stem were processed under several treatment conditions including ultrasonication treatments. The content of total saponin reached their heights at 17 hr (UGL-17) of ultrasonication treatment, followed by 16 hr (UGL-16) and 7 hr (UGL-7) of ultrasonication treatment at $100^{\circ}C$. UGL-17 findings show that the ginseng leaf and stem that had been processed with ultrasonication for 17 hours peaked in the level of Rg2, Rg3 and Rh1. In addition, UGL-16 contained ginsenoside Rg5 at high concentrations. It is thought that such results provide basic information in preparing ginseng leaf and stem extracts with functionality enhanced.

• Journal of Ginseng Research
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• 제40권4호
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• pp.400-408
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• 2016

Background: Ginsenoside Rh2 (GRh2) is the main bioactive component in American ginseng, a commonly used herb, and its antitumor activity had been studied in previous studies. PDZ-binding kinase/T-LAK cell-originated protein kinase (PBK/TOPK), a serine/threonine protein kinase, is highly expressed in HCT116 colorectal cancer cells. Methods: We examined the effect of GRh2 on HCT116 cells ex vivo. Next, we performed in vitro binding assay and in vitro kinase assay to search for the target of GRh2. Furthermore, we elucidated the underlying molecular mechanisms for the antitumor effect of GRh2 ex vivo and in vivo. Results: The results of our in vitro studies indicated that GRh2 can directly bind with PBK/TOPK and GRh2 also can directly inhibit PBK/TOPK activity. Ex vivo studies showed that GRh2 significantly induced cell death in HCT116 colorectal cancer cells. Further mechanistic study demonstrated that these compounds inhibited the phosphorylation levels of the extracellular regulated protein kinases 1/2 (ERK1/2) and (H3) in HCT116 colorectal cancer cells. In vivo studies showed GRh2 inhibited the growth of xenograft tumors of HCT116 cells and inhibited the phosphorylation levels of the extracellular regulated protein kinases 1/2 and histone H3. Conclusion: The results indicate that GRh2 exerts promising antitumor effect that is specific to human HCT116 colorectal cancer cells through inhibiting the activity of PBK/TOPK.

• Journal of Ginseng Research
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• 제37권1호
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• pp.124-134
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• 2013

The authentication of the physico-chemical properties of ginsenosides reference materials as well as qualitative and quantitative batch analytical data based on validated analytical procedures is a prerequisite for certifying good manufacturing practice (GMP). Ginsenoside Rb1 and Rg1, representing protopanaxadiol and protopanaxatriol ginsenosides, respectively, are accepted as marker substances in quality control standards worldwide. However, the current analytical methods for these two compounds recommended by Korean, Chinese, European, and Japanese pharmacopoeia do not apply to red ginseng preparations, particularly the extract, because of the relatively low content of the two agents in red ginseng compared to white ginseng. In manufacturing fresh ginseng into red ginseng products, ginseng roots are exposed to a high temperature for many hours, and the naturally occurring ginsenoside Rb1 and Rg1 are converted to artifact ginsenosides such as Rg3, Rg5, Rh1, and Rh2 during the heating process. The analysis of ginsenosides in commercially available ginseng products in Korea led us to propose the inclusion of the (20S)- and (20R)-ginsenoside Rg3, including ginsenoside Rb1 and Rg1, as additional reference materials for ginseng preparations. (20S)- and (20R)-ginsenoside Rg3 were isolated by Diaion HP-20 adsorption chromatography, silica gel flash chromatography, recrystallization, and preparative HPLC. HPLC fractions corresponding to those two ginsenosides were recrystallized in appropriate solvents for the analysis of physico-chemical properties. Documentation of those isolated ginsenosides was achieved according to the method proposed by Gaedcke and Steinhoff. The ginsenosides were subjected to analyses of their general characteristics, identification, purity, content quantification, and mass balance tests. The isolated ginsenosides showed 100% purity when determined by the three HPLC systems. Also, the water content was found to be 0.534% for (20S)-Rg3 and 0.920% for (20R)-Rg3, meaning that the net mass balances for (20S)-Rg3 and (20R)-Rg3 were 99.466% and 99.080%, respectively. From these results, we could assess and propose a full spectrum of physico-chemical properties of (20S)- and (20R)-ginsenoside Rg3 as standard reference materials for GMP-based quality control.

• 한국식품조리과학회지
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• 제28권5호
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• pp.623-628
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• 2012

본 연구에서는 가정용 전자레인지를 이용하여 간편하고 신속하게 홍삼을 제조하고 제조된 홍삼의 이화학적 특성을 조사하고자 하였다. 홍삼 제조 방법은 가정용 전자레인지의 '해동기능' 13분(A), 가정용 전자레인지의 '조리기능' 6분(B), 가정용 전자레인지의 '해동기능' 44분(C)로 하였다. 전자레인지로 제조된 홍삼의 외관, 분말의 색, 사포닌 조성, 홍삼 제조 시 용출된 사포닌의 양 등을 기존의 일반 홍삼과 비교하였다. 가정용 전자레인지의 '조리기능' 6분(B)과 '해동기능' 44분(C)으로 만든 홍삼은 일반 홍삼과 유사한 색을 가졌다. 전자레인지에 의한 홍삼 제조 시에는 기존 일반 홍삼 제조에 비해 사포닌 손실이 거의 없었다. 전자레인지 홍삼의 총페놀 함량은 일반 홍삼과 유사하였으며, 전자레인지 홍삼의 진세노사이드 함량은 일반 홍삼보다 높았다. 전자레인지 홍삼(A, B)의 진세노사이드 $Rg_1$, Re, Rf, $Rg_2+Rh_1$, $Rb_1$, Rc, $Rb_2$, $Rb_3$, Rd, $Rg_3$ 함량은 일반 홍삼보다 높았으며, 해동 기능 44분의 홍삼(C)은 진세노사이드 $Rg_1$, Re, $Rg_2+Rh_1$, Rc, $Rb_2$, $Rb_3$, Rd, $Rg_3$의 값이 일반 홍삼보다 높았다. 본 연구에서는 가정용 전자레인지를 이용하여 신속하고 간편하게 고기능성의 홍삼을 만드는 방법을 살펴보았고 이로써 인삼 소비 증진 등이 기대된다.

• Journal of Ginseng Research
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• 제33권4호
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• pp.324-329
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• 2009

G-Rh1에 의한 단핵구 세포주인 U937 세포의 유착조절 능을 조사하여 다음과 같은 결론을 얻었다. 1. G-Rh1은 CD29 항체 (MEM101A) 처리에 의해 유도된 세포-세포간 유착현상을 유의적으로 억제하였다. 2. G-Rh1은 fibronectin처리에 의해 유도된 U937 세포-fibronectin간 유착현상을 유의적으로 억제하였다. 3. G-Rh1은 CD29의 세포표면 발현 수준을 유의적으로 감소시켰다. 최근 활발히 G-Rh1의 약리작용 연구들이 진행되고 있다. 특별히, 본 화합물은 항알러지, 피부염증질환 치료효과, 항암효과 및 여성호르몬 유사기능 등이 있는 것으로 보고된 바있다. CD29-매개성 세포유착과정이 다양한 염증과정, 알러지 반응 및 암세포의 이동 및 전이성과 관련이 있다는 관점에서 볼 때 본 연구결과는 이들 약물이 갖는 치료기전의 하나가 CD29 기능저해에서 비롯될 수 있음을 제시한다고 하겠다. 이후 G-Rh1의 억제 기능에 관한 기전 이해를 위해 세포유착 유도 신호전달 단백질의 활성을 포함한 다양한 분자적 수준에서의 추가적인 실험들을 진행하고 한다.

• Archives of Pharmacal Research
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• 제27권11호
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• pp.1136-1140
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• 2004

Red ginseng and fermented red ginseng were prepared, and their composition of ginsenosides and antiischemic effect were investigated. When ginseng was steamed at 98-$100{\circ}C$ for 4h and dried for 5h at $60{\circ}C$, and extracted with alcohol, its main components were ginsenoside $Rg_3$ > ginsenoside $Rg_1$> ginsenoside $Rg_2$. When the ginseng was suspended in water and fermented for 5 days by previously cultured Bifidobacterium H-1 and freeze-dried (fermented red ginseng), its main components were compound K > ginsenoside $Rg_3{\geq}$ ginsenoside $Rg_2$. Orally administered red ginseng extract did not protect ischemia-reperfusion brain injury. However, fermented red ginseng significantly protected ischemica-reperfusion brain injury. These results suggest that ginsenoside Rh2 and compound K, which was found to be at a higher content in fermented red ginseng than red ginseng, may improve ischemic brain injury.

• 한국식품과학회지
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• 제41권1호
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• pp.77-81
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• 2009

Streptozotocin-유도 당뇨병 모델을 이용하여 흑삼의 혈당강하작용에 대한 효과를 연구하고 증포과정에 따른 ginsenoside의 조성 변화를 분석하였다. 증포별 ginsenoside의 정량분석 결과, 증포 횟수가 증가할수록 조사포닌의 총량과 대부분의 ginsenoside는 줄어든 반면 ginsenoside-$Rg_3$ 함량 및 PD/PT 비율은 증가하는 경향을 보였는데, 이러한 조성은 다른 인삼제품과는 차별성 있는 특징이다. 또한, streptozotocin-당뇨병 유발쥐를 이용한 모델실험에서 흑삼추출물의 투여는 고혈당 준위(300 mg/dL이상)를 정상 수준(102 mg/dL)으로 감소시켰으며, 혈당 수준의 감소는 glycolysis나 HMT shunt 및 지방산 합성에 관여하는 glucokinase, glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, acetyl CoA carboxylase 등의 효소의 활성증대에 기인하는 것으로 판단되었다. 이러한 결과는 흑삼추출물의 유의적인 혈당강하 효과를 입증하며 이에 따라 신약이나 식이보조제의 항당뇨물질로서의 이용가능성을 시사한다.