Effect of Ginsenoside Rc on the Pharmacokinetics of Mycophenolic Acid, a UGT1A9 Substrate, and its Glucuronide Metabolite in Rats

( So-young Park ),( Ji-hyeon Jeon ),( Su-nyeong Jang ),( Im-sook Song ),( Kwang-hyeon Liu ) 한국질량분석학회 2021 Mass spectrometry letters Vol.12 No.2

Previous in vitro studies have demonstrated that ginsenoside Rc inhibits UGT1A9, but there are no available data to indicate that ginsenoside Rc inhibits UGT1A9 in vivo. The effect of single and repeated intravenous injection of ginsenoside Rc was evaluated on the pharmacokinetics of mycophenolic acid. After injection of ginsenoside Rc (5 mg/kg for one day or 3 mg/kg for five days), 2-mg mycophenolic acid was intravenously injected, and the pharmacokinetics of mycophenolic acid and mycophenolic acid-β-glucuronide were determined. Concentrations of mycophenolic acid and its metabolite from rat plasma were analyzed using a liquid chromatography-triple quadrupole mass spectrometry. Single or repeated pretreatment with ginsenoside Rc had no significant effects on the pharmacokinetics of mycophenolic acid (P > 0.05): The mean difference in maximum plasma concentration (C_max) and area under the concentration-time curve (AUC_inf) were within 0.83- and 0.62-fold, respectively, compared with those in the absence of the ginsenoside Rc. These results indicate that ginsenoside Rc has a negligible effect on the disposition of mycophenolic acid in vivo despite in vitro findings indicating that ginsenoside Rc is a selective UGT1A9 inhibitor. As a result, ginsenoside Rc has little possibility of interacting with drugs that are metabolized by UGT1A9, including mycophenolic acid.

Ginsenosides Attenuate Formalin-Induced Pains Through Spinal and Supraspinal Sites

Yoon, So-Rah,Park, Seok,Jung, Se-Yeon,Kim, Seok-Chang,Ko, Sung-Ryong,Nam, Ki-Yeul,Nah, Seung-Yeol The Korean Society of Ginseng 2000 Journal of Ginseng Research Vol.24 No.3

In previous studies we have demonstrated that several individual ginsenosides such as Rc, Rd, Re and Ri relieves formalin-induced pain following systemic treatment. But it is unknown where these single ginsenosides induce antinociception. We investigated the antinoiceptive effect of four individual ginsenosides on formalin-induced pain after intrathecal (i.t.), intracereventricular (i.c.v.), or subcutaneous (s.c.) administration using mice. We found that ginsenoside Rc, Rd, and Re except Rf attenuated both acute and tonic phase of pain. Ginsenoside Rf attenuated only tonic phase of pain after i.t. administration. The ED$\_$50/ was 1.0 (0.55∼l.75 mg/kg) for Rc, 1.15 (0.6∼2.25 mg/kg) for Rd, and 8.9 (3.9∼20.5 mg/kg) for Re in acute phase of pain. The ED$\_$50/ was 0.3 (0.1∼0.85 mg/kg) for Rc, 0.6 (0.35∼l.1 mg/kg) for Rd, 2.45 (1.25∼4.65 mg/kg) for Re, and 1.9 (1.5∼4.25 mg/kg) for Rf in tonic phase of pain. We also found that ginsenoside Rc, Rd, Re, and Rf after i.c.v. administration attenuated both acute and tonic phase of pain. The ED5o for acute phase of pain was 0.9 (0.55∼l.4mg/kg) for Rc, 0.9 (0.45∼1.7 mg/kg) for Rd, 0.93 (0.5∼l .75 mg/kg) for Re, and 1.85 (0.95∼3.5 mg/kg) for Rf. The ED$\_$50/ for tonic phase of pain was 0.7 (0.45∼1.05 mg/kg) for Rc,1.25 (0.7∼2.2 mg/kg) for Rd, 0.85 (0.45∼1.6 mg/kg) for Re, and 0.8 (0.4∼1.45 mg/kg) for Rf. Thus, the order of the analgesic potency was Rc$\geq$Rd>Re>Rf in both i.t. and i.c.v. administration routes. However, s.c. pretreatment of four ginsenosides did not reduce formalin-induced pain. These results suggest that analgesic effect of ginsenosides is achieved through spinal or supraspinal site(s) in formalin test. 앞의 연구에서 우리는 진세노사이드 Rc, Rd, Re 및 Rr를 복강내 전 처리할 경우 포르말린으로 유도된 통증을 억제하다는 것을 보고하였다 그러나 이러한 진세노사이드가 어느 위치에서 항통증작용을 발휘하는가에 대하여서는 아직 알려지지 않고 있다. 본 연구에서는 이들 진세노사이드를 뇌실내, 척수강내 혹은 피하내 전처리한 다음 포르밀린에 의하여 유도되는 통증이 어느 위치에서 억제되는가를 연구하였다. 연구 결과 이들 진세노사이드는 척수강내 전처리할 경우 포르말린에 의하여 유도되는 통증을 억제하는 것으로 나타났다. 급성 통증 phase에서 ED$_{50}$는 Rc가 1.0 (0.SS~l.75mg/kg)이었고, Rd가 1.15 (0.6~2.25 mg/kg)이었고, Re가 8.9(3.9~20.5 mg/kg)이었다. 지속성 통증 phase에서 ED$_{50}$는 Rc가0.3 (0.1~0.85 mg/kg)이었고, Rd가 0.6 (0.35~l.1 mg/kg)이었고, Re가 2.45 (1.2s~4.65 mg/kg)이었고, Rf가 1.9(1.5~4.25 mg/kg)인 것으로 나타났다. 또한 뇌실내 전처리할 경우에도 이들 진세노사이드들은 포르말린에 의하여 유도되는 통증을 억제하였다. 급성통증 phase에서 ED$_{50}$는 Rc가 0.9 (0.55~l.4 mg/kg)이었고, Rd가0.9 (0.45~l.7 mg/kg)이었고, Re가 0.93 (0.Sol.75 mg/kg), Rf가1.85 (0.95~3.5 mg/kg)인 것으로 나타났다. 지속성 통증 phase에서는 ED$_{50}$는 Rc가 0.7 (0.45~l.05 mg/kg)이었고, Rd가 1.25(0.7~2.2 mg/kg)이었고, Re가 0.85 (0.45~l.6 mg/kg)이었고, H의 경우에는 0.8 (0.4~l.45 mg/kg) 이었다. 항통증 효능 potency는 두 가지 투여 경로에 있어서 Rc$\geq$Rd>Re>Rf인 것으로 나타났다. 그러나, 피하내 주사는 포르말린에 의하여 유도되는 통증을 억제하지 않은 것으로 나타났다. 이러한 연구 결과는 진세노사이드에 의한 항통증 작용은 척수 수준 및 척수위 수준에서 이루어진다는 것을 보여주고 있다.보여주고 있다.

Influence of Ginsenosides on the Kainic Acid-Induced Seizure Activity in Immature Rats

Park, Jin-Kyu,Jin, Sung-Ha,Choi, Keum-Hee,Ko, Ji-Hun,Baek, Nam-In,Choi, Soo-Young,Cho, Sung-Woo,Choi, Kang-Ju,Nam, Ki-Yeul Korean Society for Biochemistry and Molecular Biol 1999 Journal of biochemistry and molecular biology Vol.32 No.4

We studied the effects of ginsenosides in immature rats based upon the previous results that ginseng has a suppressive or anticonvulsive activity. To examine the suppressive effect of ginsenosides on kainic acid-induced seizures, the severities and frequencies were observed for 4 h after injection of kainic acid (KA; i.p., 2 mg/kg b.w.) using 10-day-old male Sprague-Dawley rats ($22{\pm}2\;g$). Protopanaxadiol saponins such as ginsenoside-Rb1 (Rb1), ginsenoside-Rb2 (Rb2), ginsenoside-Rc (Rc), and ginsenoside-Rd(Rd) generally reduced the seizure activities while protopanaxatriol saponins such as ginsenoside-Rg1 (Rg1) and ginsenoside-Re (Re) rather increased stereotypic "paddling-like" movements. When vinyl-GABA (v-G) was injected together with Rb1 or Rc, KA-induced seizure severities were additionally reduced only by the injection of Rc, but not by Rb1. The level of gamma isozyme of protein kinase C (PKC-${\gamma}$) in the hippocampus increased about three times as much as that of normal rats at 4 h after KA injection. The increased level of PCK-${\gamma}$ by KA was significantly reduced to about 35% by the coinjection with v-G alone, but it was not changed by v-G together with Rb1 or Rc. The increased level of PKC-${\gamma}$ at 4 h after injection of KA was not consistent with the reduction of seizure severities between Rb1 and Rc. These results suggest that Rc and Rb1 may reduce seizure severity independent of PKC-${\gamma}$ levels, and Rc may additionally act with v-G regarding the GABA metabolism during the stage of KA-induced seizures in the immature rats.

Ginsenosides Rc, as a novel SIRT6 activator, protects mice against high fat diet induced NAFLD

Zehong Yang,Yuanyuan Yu,Nannan Sun,Limian Zhou,Dong Zhang,HaiXin Chen,Wei Miao,Weihang Gao,Canyang Zhang,Changhui Liu,Xiaoying Yang,Xiaojie Wu,Yong Gao The Korean Society of Ginseng 2023 Journal of Ginseng Research Vol.47 No.3

Background: Hepatic lipid disorder impaired mitochondrial homeostasis and intracellular redox balance, triggering development of non-alcohol fatty liver disease (NAFLD), while effective therapeutic approach remains inadequate. Ginsenosides Rc has been reported to maintain glucose balance in adipose tissue, while its role in regulating lipid metabolism remain vacant. Thus, we investigated the function and mechanism of ginsenosides Rc in defending high fat diet (HFD)-induced NAFLD. Methods: Mice primary hepatocytes (MPHs) challenged with oleic acid & palmitic acid were used to test the effects of ginsenosides Rc on intracellular lipid metabolism. RNAseq and molecular docking study were performed to explore potential targets of ginsenosides Rc in defending lipid deposition. Wild type and liver specific sirtuin 6 (SIRT6, 50721) deficient mice on HFD for 12 weeks were subjected to different dose of ginsenosides Rc to determine the function and detailed mechanism in vivo. Results: We identified ginsenosides Rc as a novel SIRT6 activator via increasing its expression and deacetylase activity. Ginsenosides Rc defends OA&PA-induced lipid deposition in MPHs and protects mice against HFD-induced metabolic disorder in dosage dependent manner. Ginsenosides Rc (20mg/kg) injection improved glucose intolerance, insulin resistance, oxidative stress and inflammation response in HFD mice. Ginsenosides Rc treatment accelerates peroxisome proliferator activated receptor alpha (PPAR-α, 19013)-mediated fatty acid oxidation in vivo and in vitro. Hepatic specific SIRT6 deletion abolished ginsenoside Rc-derived protective effects against HFD-induced NAFLD. Conclusion: Ginsenosides Rc protects mice against HFD-induced hepatosteatosis by improving PPAR-α-mediated fatty acid oxidation and antioxidant capacity in a SIRT6 dependent manner, and providing a promising strategy for NAFLD.

인삼이 생쥐에서 스트레스시에 혈중 Corticosterone 농도에 미치는 영향

김도훈,민성길,손봉기,이상규,송동근 대한신경정신의학회 2002 신경정신의학 Vol.41 No.3

연구목적: 본 연구는 뇌실내 주사 스트레스와 부동 스트레스를 가했을때 인삼 사포닌 및 그 구성성분이 혈중 corticosterone에 미치는 영향을 알아보고 이런 효과에 nitric oxide가 관련되어 있는지를 조사하고자 하였다. 방 법: 스트레스를 가했을 때 인삼의 시상하부-뇌하수체-부신(hypothalamo-pituitary-adrenal, HPA) 축 에 대한 영향을 조사하기 위해서 다양한 농도의 인삼 총 사포닌과 ginsenoside Rb1, Rb2, Rc, Rd, Re, Rf, Rg1, Rg3(S), Rg3(R)을 뇌실내 주사 또는 복강내 주사 하였다. 뇌실내 주사 스트레스를 가했을 때는 스트레스를 준 후에 30분이 지나서 혈액을 채취하여 혈중 corticosterone의 농도를 측정하였다. 부동스트레스시에는 30분 동안 부동스트레스를 준 후 바로 혈액을 채취하였다. 스트레스를 가했을 때에 시상하부-뇌하수체-부신 축에 미치는 인삼의 효과에서 nitric oxide와의 관련 작용을 보기 위하여 L-NAME와 ginsenosides을 동시에 뇌실내 주사 또는 복강내 주사하고 스트레스 가한후 혈중 corticosterone의 농도를 측정하였다. 결 과: 1) 인삼 총 사포닌, ginsenoside Rg3(S) 및 Rc는 뇌실내 주사시 혈중 corticosterone의 증가를 억제 하였고 이러한 효과는 nitric oxide에 의해 상쇄 되었다 2) 인삼 총 사포닌, ginsenoside Rc의 복강내 투여는 부동 스트레스에 의한 혈중 corticosterone의 증가를 억제하였다. 그러나 뇌실내 주사시와는 달리 부동 스트레스시 복강으로 투여된 ginsenoside Rg3(S)은 혈중 corticosterone의 증가를 억제하지 않았다. 복강으로 투여된 인삼 총 사포닌의 부동 스 트레스시 혈중 corticosterone 증가억제 작용은 nitric oxide 생성억제제인 NG-Nitro-L-arginine methyl ester(L-NAME)에 의하여 상쇄되지 않았는데 이는 nitric oxide가 관련되지 않음을 시사한다. 결 론: 본 연구의 결과는 인삼 사포닌이 스트레스시에 시상하부-뇌하수체-부신 축을 억제하는 작용이 있고, 인삼 사포닌의 성분 및 스트레스의 종류, 투여 경로에 따라서 그 작용 기전이 다르다는 점을 제시한다. Objectives:The present study was designed to investigate the effect of ginseng saponin and its major active metabolite on the HPA axis under acute stress-i.c.v. injection stress, and immobilization stress, and to examine whether nitric oxide is involved in the mechanism of ginseng saponin on the HPA axis under acute stress. Methods:In the experiment to study the effect of ginseng on HPA axis during stress, various dose of GTS were injected intracerebroventricularly(i.c.v.) or intraperitoneally(i.p.). Plasma corticosterone levels were measured 30 min after the i.c.v. injection stress. Immobilization stress was applied for 30 min and then blood was cellected for the assays of plasma corticosterone levels immediately after the completion of immobilization stress. To determine the active ginsenosides that can affect the stressinduced plasma corticosterone levels, various dose of each gisendosides(Rb1, Rb2, Rc, Re, Rf, Rg1, 20(S)-Rg3, and 20(R)-Rg3) were injected i.c.v. or i.p.. In the experiment to determine the involvement of the nitric oxide in the inhibitory effect of ginseng on the HPA, NG-Nitro-L-arginine methyl ester(L-NAME) and ginsenosides were coadministered i.c.v. or i.p., and plasma corticosterone levels were measured 30 min after stress was applied. Results:First, the present study showed that ginseng total saponin, ginsenoside Rg3(S form), and ginsenoside Rc administered i.c.v. attenuated the intracerebroventricular injection stress-induced increase in plasma corticosterone levels, and these effects were removed by nitric oxide co-injection. Second, ginseng total saponin and ginsenoside Rc administered i.p. attenuated the immobilization stress-induced increase in plasma corticosterone levels, but ginsenoside Rg3(S form) did not attenuate the immobilization stress-induced increase in plasma corticosterone levels. The attenuative effects of ginseng total saponin and ginsenoside Rc in the immobilization stress-induced increase in plasma corticosterone levels were not affected by L-NAME co-injection. Conclusion:This study suggests that ginseng saponin attenuated stress-induced increase in plasma corticosterone levels and these effects were mediated by different mechanisms according to the components of ginseng saponin, and routes of administration

Influence of Ginsenosides on the Kainic Acid-Induced Seizure Activity in Immature Rats

Park, Jin-Kyu,Jin, Sung-Ha,Choi, Keum-Hee,Ko, Ji-Hun,Baek, Nam-In,Choi, Soo-Young,Cho, Sung-Woo,Choi, Kang-Ju,Nam, Ki-Yeula The Korea Science and Technology Center 1999 BMB Reports Vol.32 No.4

We studied the effects of ginsenosides in immature rats based upon the previous results that ginseng has a suppressive or anticonvulsive activity. To examine the suppressive effect of ginsenosides on kainic acid-induced seizures, the severities and frequencies were observed for 4 h after injection of kainic acid (KA; i.p., 2 mg/kg b.w.) using 10-day-old male Sprague-Dawley rats (22 ±2g). Protophanaxadiol saponins such as ginsenoside-Rb1 (Rb1), ginsenoside-Rb2 (Rb2), ginsenoside-Rc (Rc), and ginsenoside-Rd (Rd) generally reduced the seizure activities while protopanaxatriol saponins such as ginsenoside-Rg1 (Rg1) and ginsenoside-Re (Re) rather increased stereotypic "paddling-like" movements. When vinyl-GABA (v-G) was injected together with Rb1 or Rc, KA-induced seizure severities were additionally reduced only by the injection of Rc, but not by Rb1. The level of gamma isozyme of protein kinase C (PKC-γ) in the hippocampus increased about three times as much as that of normal rats at 4 h after KA injection. The increased level of PKC-γ by KA was significantly reduced to about 35% by the coinjection with v-G alone, but it was not changed by v-G together with Bb1 or Rc. The increased level of PKC-γ at 4 h after injection of KA was not consistent with the reduction of seizure severities between Rb1 and Rc. These results suggest that Rc and Rb1 may reduce seizure severity independent of PKC-γ levels, and Rc may additionally act with v-G regarding the GABA metabolism during the stage of KA-induced seizures in the immature rats.

Protective Effects of Ginsenosides on Cyanide-induced Neurotoxicity in Cultured Rat Cerebellar Granule Cells

Seong, yeon-Hee,Koh, Sang-Bum,Jo, Soon-Ok The Korean Society of Ginseng 2000 Journal of Ginseng Research Vol.24 No.4

흰쥐 소뇌로부터 과립신경세포를 배양하여 NaCN으로 유도되는 신경세포손상에 대한 ginsenosides의 보호효과를 검토하였다. NaCN(I~10 M)을 배양된 세포에 1시간 동안 처리하면 농도 의존적으로 신경세포사를 일으켰다. Ginsenosides(Rb$_1$, Rc, Re, Ri, Rg$_1$)(0.5, 5 $\mu\textrm{g}$/ml를 세포에 전처치하면 10 mM NaCN으로 유도되는 세포사가 현저히 감소되었다. Rb$_1$과 Rc(5$\mu\textrm{g}$/ml)는 5 mM NaCN에 의하여 배양액 중으로 유리되는 glutamate의 증가를 현저히 억제하였으며, 1 mM N3CN에 의하여 유발되는 세포내 $Ca^{2+}$농도의 증가를 억제하였다. NaCN으로 유발되는 세포독성은 또한 MK-801, verapamil, NAME에 의하여도 억제되었다. 따라서, NaCN으로 유도되는 신경세포사는 glutamate release를 통한 NMDA수용체의 활성화와 그에 따른 $Ca^{2+}$의 세포내유입에 의한 것임을 알수 있고, ginsenosides, 특히 Rb$_1$과 Rc는 $Ca^{2+}$의 유입을 억제하므로서 NaCN에 의한 신경세포사를 억제하는 것으로 생각된다. Effects of ginsenosides on NaCN-induced neuronal cell death were studied in cultured rat cerebellar granule cells. NaCN produced a concentration-dependent (1-10 mM) reduction of cell viability (measured by frypan blue exclusion test), that was blocked by N-methyl-D-aspartate receptor antagonist (MK-801) and L-type Ca$\^$2+/ channel blocker (verapamil). Pretreatment with ginsenosides (Rb$_1$, Rc, Re, Rf and Rg$_1$) significantly decreased the neuronal cell death in a concentration range of 0.5∼5$\mu\textrm{g}$/ml. Ginsenosides Rb$_1$ and Rc (5 $\mu\textrm{g}$/ml) inhibited glutamate release into medium induced by NaCN (5 mM). NaCN (1 mM)-induced increase of [Ca$\^$2+/], was significantly inhibited by the pretreatment of Rb$_1$ and Rc (5 $\mu\textrm{g}$/ml). Other ginsenosides caused relatively little inhibition on the elevation of glutamate release and of (Ca$\^$2+/). These results suggest that the NaCN-induced neurotoxicity was related to a series of cell responses consisting of glutamate release and [Ca$\^$2+/]i elevation via glutamate (NMDA and kainate) receptors and resultant cell death, and that ginsenosides, especially Rb$_1$ and Rc, prevented the neuronal cell death by the blockade of the NaCN-induced Ca$\^$2+/influx.

Development of a rapid and convenient method to separate eight ginsenosides from Panax ginseng by high-speed counter-current chromatography coupled with evaporative light scattering detection

Omer Shehzad,In Jin Ha,Sung Ho Son,Youmie Park,Yeong Shik Kim 한국당과학회 2011 한국당과학회 학술대회 Vol.2011 No.1

Ginsenosides are well-known major components isolated from the radix of Panax ginseng C. A. Meyer, known as Korean ginseng, having diverse biological activities. They have recently gained much attention for their biomedical applications. In the present work, a fast and simple method for the separation and purification of 8 ginsenosides from Panax ginseng by counter current chromatography coupled with evaporative light scattering detector (CCC-ELSD) was successfully established. The crude samples for CCC separation were first purified from ginseng extract using a macroporous resin. The extract was loaded onto a Diaion-HP20 column and fractionated by methanol and water gradient elution. The ginsenosides-triols and diols fractions were subsequently eluted with 65% & 80% methanol and water gradient elution, respectively. Furthermore, these two fractions were separated by CCC-ELSD. The two phase solvent system used for separation was composed of chloroform-methanol-water-isopropanol at a volume ratio of 4:3:2:1. Each fraction obtained was collected and dried, which yielded eight ginsenosides namely, Rg1, Re, Rf, Rh1, Rb1, Rc, Rb2 & Rd. The purity of these ginsenosides was more than 97% assessed by HPLC-ELSD system, and their structures were characterized by electrospray-ionization mass spectrometry (ESI-MS), 1H NMR and 13C NMR spectroscopy. This is the first report regarding the CCC separation of ginsenosides Rh1, Rb2 & Rc from Panax ginseng.

Production of Ginsenoside Rd from Ginsenoside Rc by α-L-Arabinofuranosidase from Caldicellulosiruptor saccharolyticus

( Kyung Chul Shin ),( Gi Woong Lee ),( Deok Kun Oh ) 한국미생물 · 생명공학회 2013 Journal of microbiology and biotechnology Vol.23 No.4

Ginsenoside Rd was produced from ginsenoside Rc using a thermostable recombinant α-L-arabinofuranosidase from Caldicellulosiruptor saccharolyticus. The optimal reaction conditions for the production of ginsenoside Rd from Rc were pH 5.5, 80℃, 227 U enzyme/ml, and 8.0 g/l ginsenoside Rc in the presence of 30% (v/v) n-hexane. Under these conditions, the enzyme produced 7.0 g/l ginsenoside Rd after 30 min, with a molar yield of 100% and a productivity of 14g l-1 h-1. The conversion yield and productivity of ginsenoside Rd are the highest reported thus far among enzymatic transformations.

Ginsenoside Rc and Re Stimulate c-Fos Expression in MCF-7 Human Breast Carcinoma Cells

YoungJooLee,YoungRanJin,임원청,SangMiJi,JungYoonCho,반재준,이승기 대한약학회 2003 Archives of Pharmacal Research Vol.26 No.1

We have found that ginsenoside Rc and Re induce c-fos in MCF-7 human breast carcinoma cells at both the mRNA and protein levels. However, neither ginsenoside activated the expression of reporter gene under the control of AP-1/TPA response elements. We have also examined the possibility that ginsenoside Rc and Re act by binding to intracellular steroid hormone receptors that act as transcriptional factors in the nucleus in inducing c-fos mRNA in MCF7 human breast carcinoma cells. However, ginsenoside Rc and Re did not bind to glucocorticoid, androgen, estrogen, or retinoic acid receptors as examined by the transcription activation of the luciferase reporter genes in CV-1 cells that were transiently transfected with the corresponding steroid hormone receptors and hormone responsive luciferase reporter plasmids. These data demonstrate that ginsenoside Rc and Re act via other transcription factors and not via estrogen receptor in c-Fos expression.