Korean Red Ginseng inhibits apoptosis in neuroblastoma cells via estrogen receptor β-mediated PI3K/AKT signaling – Accepted Manuscript

Abstract: Background: Ginseng has been shown to exert anti-stress effects both in vitro and in vivo. However, the effects of ginseng on stress in brain cells are not well understood. In this study, we investigated how Korean Red inseng (KRG) controls hydrogen peroxide-induced apoptosis via regulation of PI3K/AKT and estrogen receptor β signaling.Methods: Human neuroblastoma SK-N-SH cells were pre-treated with KRG and subsequently exposed to H2O2. The ability of KRG to inhibit oxidative stress-induced apoptosis was assessed in MTT cytotoxicity assays. Apoptotic protein expression was examined by western blot analysis. The roles of ER-β, PI3K, and p-AKT signaling in KRG regulation of apoptosis were studied using small interfering RNAs and/or target antagonists.Results: Pre-treating SK-N-SH cells with KRG decreased expression of the pro-apoptotic proteins p-p53 and caspase-3, but increased expression of the anti-apoptotic protein BCL2. KRG pre-treatment was also associated with increased ERβ, PI3K, and p-AKT expression. Conversely, ER-β inhibition with siRNA or inhibitor treatment increased p-p53 and caspase-3 levels, but decreased BCL2, PI3K, and p-AKT expression. Moreover, inhibition of PI3K/AKT signaling diminished p-p53 and caspase-3 levels, but increased BCL2 expression.Conclusion: Collectively, our data indicate that KRG represses oxidative stress-induced apoptosis by enhancing PI3K/AKT signaling via up-regulation of ER-β expression.

Soil properties of cultivation sites for mountain-cultivated ginseng at the local level – Accepted Manuscript

Abstract: Background: Identifying suitable sites for growing mountain-cultivated ginseng are an area of concern for ginseng producers. This study was conducted to evaluate the soil properties of cultivation sites for mountain-cultivated ginseng in Hamyang-gun, which is known to be one of the most famous areas for mountain-cultivated ginseng in Korea.Methods: The sampling plots from 30 sites were randomly selected on or near the center of the ginseng growing sites in July and August 2009. Soil samples for the soil property analysis were collected through the top 20 cm at five randomly selected points.Results: Mountain-cultivated ginseng was grown in soils that varied greatly in soil properties on coniferous, mixed, and deciduous broadleaved stand sites of elevations between > 200 m and < 1,000 m. The soil bulk density was higher in Pinus densiflora than in Larix leptolepis stand sites and higher in the 700 m sites. Soil pH was unaffected by the type of stand sites (pH 4.35–4.55), while the high-elevation sites of > 700 m were strongly acidified, with pH 4.19. The organic carbon and total nitrogen content were lower in the P. densiflora stand sites than in the deciduous broadleaved stand sites. Available phosphorus was low in all of the stand sites. The exchangeable cation was generally higher in the mixed and low-elevation sites than in the P. densiflora and high-elevation sites, respectively.Conclusion: These results indicate that mountain-cultivated ginseng in Korea is able to grow in very acidic, nutrient-depleted forest soils.

Ginsenoside Rp1-induced apolipoprotein A-1 expression in the LoVo human colon cancer cell line – Accepted Manuscript

Abstract: Background: Ginsenoside Rp1 (G-Rp1) is a novel ginsenoside derived from ginsenoside Rk1. This compound was reported to have anti-cancer, anti-platelet, and anti-inflammatory activities. In this study, we examined the molecular target of G-Rp1’s anti-proliferative and pro-apoptotic activities.Methods: To examine the effects of G-Rp1, cell proliferation assays, propidium iodine staining, proteomic analysis by two-dimensional gel electrophoresis, immunoblotting analysis, and a knockdown strategy were employed.Results: G-Rp1 dose-dependently suppressed the proliferation of colorectal cancer LoVo cells and also increased the apoptosis of these cells. Interestingly, G-Rp1 remarkably up-regulated the protein level of apolipoprotein (Apo)-A1 in LoVo, SNU-407, DLD-1, SNU-638, AGS, KPL-4, and SK-BR-3 cells. The knockdown of Apo-A1 by its siRNA increased the levels of cleaved poly(ADP-ribose) polymerase (c-PARP) and p53 and diminished the proliferation of LoVo cells.Conclusion: These results suggest that G-Rp1 may act as an anti-cancer agent by strongly inhibiting cell proliferation and enhancing cell apoptosis through the up-regulation of Apo-A1.

Molecular mechanism of protopanaxadiol saponin fraction-mediated anti-inflammatory actions – Accepted Manuscript

Abstract: Korean Red Ginseng (KRG) is a representative traditional herbal medicine with many different pharmacological properties including anti-cancer, anti-atherosclerosis, anti-diabetic, and anti-inflammatory activities. Since only a few studies have explored the molecular mechanism of KRG-mediated anti-inflammatory activities, in this study we aimed to investigate the anti-inflammatory mechanisms of the protopanaxadiol saponin fraction (PPD-SF) of KRG using in vitro and in vivo inflammatory models. PPD-SF dose-dependently diminished the release of inflammatory mediators [nitric oxide (NO), tumor necrosis factor (TNF)-α, and prostaglandin E2 (PGE2)], and downregulated the mRNA expression of their corresponding genes [inducible NO synthase (iNOS), TNF-α, and cyclooxygenase (COX)-2], without altering cell viability. The PPD-SF-mediated suppression of these events appeared to be regulated by a blockade of p38, c-Jun N-terminal kinase (JNK), and TANK-binding kinase 1 (TBK1), which are linked to the activation of activating transcription factor 2 (ATF2) and interferon regulatory transcription factor 3 (IRF3). Moreover, this fraction also ameliorated EtOH/HCl-induced gastritis via suppression of phospho-JNK2 levels. Therefore, these results strongly suggest that the anti-inflammatory action of PPD-SF could be mediated by a reduction in the activation of p38-, JNK2-, and TBK1-linked pathways and their corresponding transcription factors (ATF2/IRF3).

Rg3-enriched Korean Red Ginseng improves vascular function in spontaneously hypertensive rats – Accepted Manuscript

Abstract: Panax ginseng has distinct and impressive health benefits, such as improved blood pressure and immune system functioning. Rg3-enriched Korean red ginseng (REKRG) isolated from Korean red ginseng contains a high percentage of Rg3. In this study, we examined the effects of REKRG on eNOS activation and adhesion molecules in endothelial cells and vascular function in rats. REKRG dose-dependently increased endothelial nitric oxide synthase (eNOS) phosphorylation and nitric oxide (NO) production in endothelial cells. In addition, REKRG markedly inhibited the TNF-α-mediated induction of intercellular adhesion molecule (ICAM)-1 and cyclooxygenase (COX)-2 expressions in endothelial cells. REKRG improved endothelium-dependent vasorelaxation in Wistar-Kyoto rat (WKY) and spontaneously hypertensive rats (SHRs) compared with controls. Furthermore, REKRG treatment for 6 weeks increased serum NO levels and reduced the mean aortic intima-media thickness compared with controls. Taken together, these results suggest that REKRG increased vascular function and improved immune system functioning. Therefore, REKRG is a very useful food for preventing or improving various cardiovascular diseases.

Transcriptome profiling and comparative analysis of Panax ginseng adventitious roots – Accepted Manuscript

Abstract: Background: Panax ginseng Meyer is a traditional medicinal plant famous for its strong therapeutic effects and serves as an important herbal medicine. To understand and manipulate genes involved in secondary metabolic pathways including ginsenosides, transcriptome profiling of P. ginseng is essential.Methods: RNA-seq analysis of adventitious roots of two P. ginseng cultivars, Chunpoong (CP) and Cheongsun (CS), was performed using Illumina HiSeq platform. After transcript was assembled, expression profiling was performed.Results: The assemblies were generated from ∼85 and ∼77 million high-quality reads from CP and CS cultivars, respectively. A total of 35,527 and 27,716 transcripts were obtained from the CP and CS assemblies, respectively. Annotation of the transcriptomes showed that approximately 90% of the transcripts had significant matches in public databases. We identified several candidate genes involved in ginsenoside biosynthesis. In addition, a large number of transcripts (17%) with different GO designations were uniquely detected in adventitious roots compared to normal ginseng roots.Conclusion: This study will provide comprehensive insight into transcriptome of ginseng adventitious roots and a way for successful transcriptome analysis and profiling of resources plants with less genomics information. The transcriptome profiling data generated in this study are available in our newly created adventitious root transcriptome database (http://im-crop.snu.ac.kr/transdb/index.php) for public use.

Korean Red Ginseng attenuates anxiety-like behavior during ethanol withdrawal in rats – Accepted Manuscript

Abstract: Background: Korean Red Ginseng is known to have anti-anxiety properties. This study was conducted to investigate the anxiolytic effects of Korean Red Ginseng extract (KRGE) during ethanol withdrawal (EW) and the involvement of the mesoamygdaloid dopamine (DA) system in it.Methods: Rats were treated with 3 g/kg/day of ethanol for 28 days, and subjected to three days of withdrawal. During EW, KRGE (20 mg/kg/day or 60 mg/kg/day, p.o.) was given to rats once a day for three days. Thirty minutes after the final dose of KRGE, anxiety-like behavior was evaluated in an elevated plus maze (EPM), and plasma corticosterone (CORT) levels were determined by a radioimmunoassay (RIA). In addition, concentrations of DA and 3, 4-dihydroxyphenylacetic acid (DOPAC) in the central nucleus of the amygdala (CeA) were also measured by high performance liquid chromatography (HPLC).Results: The EPM test and RIA revealed KRGE inhibited anxiety-like behavior and the over secretion of plasma CORT during EW. Furthermore, the behavioral effect was blocked by a selective dopamine D2 receptor antagonist (eticlopride) but not by a selective dopamine D1 receptor antagonist (SCH23390). HPLC analyses showed KRGE reversed EW-induced decreases of DA and DOPAC in a dose-dependent way. Additionally, western blotting and real-time polymerase chain reaction assays showed KRGE prevented the EW-induced reductions in tyrosine hydroxylase protein expression in the CeA and tyrosine hydroxylase mRNA expression in the ventral tegmental area.Conclusions: These results suggest KRGE has anxiolytic effects during EW by improving the mesoamygdaloid DA system.

In vitro cytotoxic activity of ginseng leaves/stems extracts obtained by subcritical water extraction – Accepted Manuscript

Abstract: Ginseng leaves/stems extract produced by subcritical water (SW) extraction at high temperature (190°C) possessed higher cytotoxic activity against human cancer cell lines than ethanol extract. SW extraction can be a great candidate for extraction of functional substance from ginseng leaves/stems.