The study was performed to evaluate the antibacterial and antiviral activities of ginseng fine root in order to search for antibacterial substances. Among 8 kinds of fermentation strains, Lactobacillus plantarum was selected based on viable cell count and antibacterial activities during incubation. Optimum conditions of ginseng fine root fermentation for L. plantarum were incubation at $35^{\circ}C$ for 48 hr in 5% ginseng fine root broth. That methanolic extract of fermented ginseng fine root broth was observed to be antibacterial and have antiviral activities. The results of paper disc method of non-fermented extract and fermented extract measured against E. coli was 11 mm and 20 mm, S. aureus was 15 mm and 22 mm, respectively. Shaking flask method was observed to inhibit the growth E. coli and S. aureus in fermented extract by 99.9%. However, antiviral activity of Feline calicivirus (FCV) was mostly activated. Fermented extract was used to investigate the compositional changes of ginsenosides on HPLC analysis. By fermentation, ginsenoside Rg1, Re and Rd were increased, with Rd showing a significant increase of 50 ${\mu}g/g$. These results suggest that ginseng fine root extract is a useful resource.
Kim, Seung Tae;Kim, Hee Jung;Jang, Su Kil;Lee, Do Ik;Joo, Seong Soo
Korean Journal of Food Science and Technology
/
v.45
no.1
/
pp.77-83
/
2013
In this study, we observed optimal conditions and suitable bacteria for the fermentation of steam-dried ginseng berry extracts (SGB) and determined antioxidant effects of the fermented extracts. Five bacteria (Lactobacillus fermentarum, L. plantarum, L. brevis, L. casei, Bacillus subtillis) were examined on their growth activities and viabilities in various culture temperatures ($25-35^{\circ}C$) and concentrations (25-100%). L. plantarum was considered to be the most suitable bacteria for the fermentation in both growth activity and viability. Moreover, the extracts fermented with L. plantarum showed more potent antioxidant efficacy in both 1,1-diphenyl-2-picrylhydrazyl radical and hydroxyl radical scavenging assay. High performance liquid chromatography analysis revealed that fermentation with L. plantarum changed the contents and components of ginsenosides. In conclusion, these data suggest that L. plantarum efficiently ferment SGB and the fermented extracts may have therapeutical values against oxidative stress and be a good candidate in adjuvant therapy where ginsenoside would be the main composition.
Background: Sun ginseng (SG), a specific formulation of quality-controlled red ginseng, contains approximately equal amounts of three major ginsenosides (RK1, Rg3, and Rg5), which reportedly has antitumor-promoting activities in animal models. Methods: MTT assay was used to assess whether SG can potentiate the anticancer activity of epirubicin or paclitaxel in human cervical adenocarcinoma HeLa cells, human colon cancer SW111C cells, and SW480 cells; apoptosis status was analyzed by annexin V-FITC and PI and analyzed by flow cytometry; and apoptosis pathway was studied by analysis of caspase-3, -8, and -9 activation, mitochondrial accumulation of Bax and Bak, and cytochrome c release. Results: SG remarkably enhances cancer cell death induced by epirubicin or paclitaxel in human cervical adenocarcinoma HeLa cells, human colon cancer SW111C cells, and SW480 cells. Results of the mechanism study highlighted the cooperation between SG and epirubicin or paclitaxel in activating caspase-3 and -9 but not caspase-8. Moreover, SG significantly increased the mitochondrial accumulation of both Bax and Bak triggered by epirubicin or paclitaxel as well as the subsequent release of cytochrome c in the targeted cells. Conclusion: SG significantly potentiated the anticancer activities of epirubicin and paclitaxel in a synergistic manner. These effects were associated with the increased mitochondrial accumulation of both Bax and Bak that led to an enhanced cytochrome c release, caspase-9/-3 activation, and apoptosis. Treating cancer cells by combining epirubicin and paclitaxel with SG may prove to be a novel strategy for enhancing the efficacy of the two drug types.
Cho, Do-Yeon;Skinner, Daniel;Zhang, Shaoyan;Lazrak, Ahmed;Lim, Dong Jin;Weeks, Christopher G.;Banks, Catherine G.;Han, Chang Kyun;Kim, Si-Kwan;Tearney, Guillermo J.;Matalon, Sadis;Rowe, Steven M.;Woodworth, Bradford A.
Journal of Ginseng Research
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v.45
no.1
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pp.66-74
/
2021
Background: Abnormal chloride (Cl-) transport has a detrimental impact on mucociliary clearance in both cystic fibrosis (CF) and non-CF chronic rhinosinusitis. Ginseng is a medicinal plant noted to have anti-inflammatory and antimicrobial properties. The present study aims to assess the capability of red ginseng aqueous extract (RGAE) to promote transepithelial Cl- secretion in nasal epithelium. Methods: Primary murine nasal septal epithelial (MNSE) [wild-type (WT) and transgenic CFTR-/-], fisher-rat-thyroid (FRT) cells expressing human WT CFTR, and TMEM16A-expressing human embryonic kidney cultures were utilized for the present experiments. Ciliary beat frequency (CBF) and airway surface liquid (ASL) depth measurements were performed using micro-optical coherence tomography (μOCT). Mechanisms underlying transepithelial Cl- transport were determined using pharmacologic manipulation in Ussing chambers and whole-cell patch clamp analysis. Results: RGAE (at 30㎍/mL of ginsenosides) significantly increased Cl- transport [measured as change in short-circuit current (ΔISC = ㎂/㎠)] when compared with control in WT and CFTR-/- MNSE (WT vs control = 49.8±2.6 vs 0.1+/-0.2, CFTR-/- = 33.5±1.5 vs 0.2±0.3, p < 0.0001). In FRT cells, the CFTR-mediated ΔISC attributed to RGAE was small (6.8 ± 2.5 vs control, 0.03 ± 0.01, p < 0.05). In patch clamp, TMEM16A-mediated currents were markedly improved with co-administration of RGAE and uridine 5-triphosphate (8406.3 +/- 807.7 pA) over uridine 5-triphosphate (3524.1 +/- 292.4 pA) or RGAE alone (465.2 +/- 90.7 pA) (p < 0.0001). ASL and CBF were significantly greater with RGAE (6.2+/-0.3 ㎛ vs control, 3.9+/-0.09 ㎛; 10.4+/-0.3 Hz vs control, 7.3 ± 0.2 Hz; p < 0.0001) in MNSE. Conclusion: RGAE augments ASL depth and CBF by stimulating Cl- secretion through CaCC, which suggests therapeutic potential in both CF and non-CF chronic rhinosinusitis.
Yang Zhang;Jiulong Ma;Shan Liu;Chen Chen;Qi Li;Meng Qin;Liqun Ren
Journal of Ginseng Research
/
v.47
no.1
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pp.106-116
/
2023
Background: Pirarubicin (THP) is an anthracycline antibiotic used to treat various malignancies in humans. The clinical usefulness of THP is unfortunately limited by its dose-related cardiotoxicity. Ginsenoside F1 (GF1) is a metabolite formed when the ginsenosides Re and Rg1 are hydrolyzed. However, the protective effects and underlying mechanisms of GF1 on THP-induced cardiotoxicity remain unclear. Methods: We investigated the anti-apoptotic and anti-oxidative stress effects of GF1 on an in vitro model, using H9c2 cells stimulated by THP, plus trigonelline or AKT inhibitor imidazoquinoxaline (IMQ), as well as an in vivo model using THP-induced cardiotoxicity in rats. Using an enzyme-linked immunosorbent test, the levels of malondialdehyde (MDA), brain natriuretic peptide (BNP), creatine kinase (CK-MB), cardiac troponin (c-TnT), lactate dehydrogenase (LDH), superoxide dismutase (SOD) and glutathione (GSH) were determined. Nuclear factor (erythroid-derived2)-like 2 (Nrf2) and the expression of Nrf2 target genes, including heme oxygenase-1 (HO-1), glutathione-S-transferase (Gst), glutamate-cysteine ligase modifier subunit (GCLM), and expression levels of AKT/Bcl-2 signaling pathway proteins were detected using Western blot analysis. Results: THP-induced myocardial histopathological damage, electrocardiogram (ECG) abnormalities, and cardiac dysfunction were reduced in vivo by GF1. GF1 also decreased MDA, BNP, CK-MB, c-TnT, and LDH levels in the serum, while raising SOD and GSH levels. GF1 boosted Nrf2 nuclear translocation and Nrf2 target gene expression, including HO-1, Gst, and GCLM. Furthermore, GF1 regulated apoptosis by activating AKT/Bcl-2 signaling pathways. Employing Nrf2 inhibitor trigonelline and AKT inhibitor IMQ revealed that GF1 lacked antioxidant and anti-apoptotic effects. Conclusion: In conclusion, GF1 was found to alleviate THP-induced cardiotoxicity via modulating Nrf2 and AKT/Bcl-2 signaling pathways, ultimately alleviating myocardial oxidative stress and apoptosis.
Background: Lung inflammation occurs in many lung diseases, but has limited effective therapeutics. Ginseng and its derivatives have anti-inflammatory effects, but their unstable physicochemical and metabolic properties hinder their application in the treatment. Panaxadiol (PD) is a stable saponin among ginsenosides. Inhalation administration may solve these issues, and the specific mechanism of action needs to be studied. Methods: A mouse model of lung inflammation induced by lipopolysaccharide (LPS), an in vitro macrophage inflammation model, and a coculture model of epithelial cells and macrophages were used to study the effects and mechanisms of inhalation delivery of PD. Pathology and molecular assessments were used to evaluate efficacy. Transcriptome sequencing was used to screen the mechanism and target. Finally, the efficacy and mechanism were verified in a human BALF cell model. Results: Inhaled PD reduced LPS-induced lung inflammation in mice in a dose-dependent manner, including inflammatory cell infiltration, lung tissue pathology, and inflammatory factor expression. Meanwhile, the dose of inhalation was much lower than that of intragastric administration under the same therapeutic effect, which may be related to its higher bioavailability and superior pharmacokinetic parameters. Using transcriptome analysis and verification by a coculture model of macrophage and epithelial cells, we found that PD may act by inhibiting TNFA/TNFAR and IL7/IL7R signaling to reduce macrophage inflammatory factor-induced epithelial apoptosis and promote proliferation. Conclusion: PD inhalation alleviates lung inflammation and pathology by inhibiting TNFA/TNFAR and IL7/IL7R signaling between macrophages and epithelial cells. PD may be a novel drug for the clinical treatment of lung inflammation.
Most neurodegenerative diseases are known to be influenced by oxidative stress. We investigated the anti-oxidative activity of the concentrate of fermented wild ginseng root culture (HLJG0701) containing ginsenosides Rg5 and Rk1. HLJG0701 showed effective DPPH and ABTS radical scavenging ability ($IC_{50}$: 16- and 4-fold dilution, respectively) and was inhibited dose-dependently by the $FeSO_4$-induced lipid peroxidation group (8- and 4-fold dilution: 2.3 and 1.5 nM, respectively). In MTT and LDH assays, 8-, 16-, 32- and 64-fold diluted HLJG0701 significantly increased cell viability by 70, 53, 35, and 26%, respectively. LDH released by HLJG0701 was reduced 1.3-fold with 8-fold diluted HLJG0701 compared to the $H_2O_2$-treated control. In addition, the inhibitory effect of HLJG0701 on oxidative stress in PC12 cells was confirmed by DCF-DA analysis (16-, 4-fold diluted HLJG0701: 50 and 68% ROS inhibition, respectively), TBARS (16- and 4-fold diluted HLJG0701: 50.7 and 46.5% inhibition, respectively), GPx (16- and 4-fold diluted HLJG0701: 133.3 and 227.3% release, respectively), and SOD analysis (16- and 4-fold diluted HLJG0701: 118.2 and 218.2% release, respectively). These results suggested that HLJG0701 protects neuronal cells by its anti-oxidative effects and hence can be a potential preventive material against neurodegenerative diseases.
In order to reduce the bitter taste and improve the bioavailability of red ginseng extract(RGE), inclusion complexes (RGE-CD) of the extract with ${\alpha}-,\;{\beta}-,\;{\gamma}$-cyclodextrin were prepared and studied for their sensory quality and bioavailability compared to RGE. By complexation, the bitter taste-reducing efficacies of ${\alpha}$-CD and ${\beta}$-CD were much lower than that of ${\gamma}$-CD. In comparative sensory analysis for the bitter taste, RGE-${\gamma}$-CD10, prepared using 10%(w/w) of ${\gamma}$-CD, showed a score of 1.93(decreased by about 78%) compared to RGE as the control. In addition, in sensory analysis for flavor, RGE-${\gamma}$-CD10showed a score of 5.60. Upon increasing the amount of ${\gamma}$-CD to 15%(w/w) and 20%(w/w), respectively, the bitter taste of RGE-${\gamma}$-CD was removed and the flavor of RGE disappeared(scores of 2.67 and 1.67, respectively). Therefore RGE-${\gamma}$-CD10 was chosen as an optimum. The same dosages of RGE and RGE-${\gamma}$-CD10 were orally administered to SD(Sprague-Dawley) rats on a saponin basis, and the plasma concentrations of ginsenoside Rg1 and Rb1 were measured over time to estimate the average AUC(area under the plasma concentration versus time curve) of the ginsenosides. After the oral administration, there were no significant differences in the AUC values of the RGE and RGE-${\gamma}$-CD 10 groups for ginsenoside Rg1. However, AUC values for ginsenoside Rb1 were $25.8{\mu}g{\cdot}hr/mL$ in the RGE group and $81.5{\mu}g{\cdot}hr/mL$ in the RGE-${\gamma}$-CD 10 group, respectively. Therefore, the bioavailability of ginsenoside Rb1 in the RGE-${\gamma}$-CD 10 group was significantly higher by up to 315% compared with that in the RGE group(p = 0.0029). These results show that the bitter taste of RGE can be simultaneously removed by the complexation of RGE and ${\gamma}$-CD(RGE-${\gamma}$-CD) along with increased bioavailability.
Nicotinic acetylcholine receptors (nAChRs) play important roles in nervous system functions and are involved in a variety of diseases. We previously demonstrated that ginsenosides, the active ingredients of Panax ginseng, inhibit subsets of nAChR channel currents, but not ${\alpha}7$, expressed in Xenopus laevis oocytes. Mutation of the highly conserved Leu247 to Thr247 in the transmembrane domain 2 (TM2) channel pore region of ${\alpha}7$ nAChR induces alterations in channel gating properties and converts ${\alpha}7$ nAChR antagonists into agonists. In the present study, we assessed how point mutations in the Leu247 residue leading to various amino acids affect 20(S)-ginsenoside $Rg_3$ ($Rg_3$) activity against the ${\alpha}7$ nAChR. Mutation of L247 to L247A, L247D, L247E, L247I, L247S, and L247T, but not L247K, rendered mutant receptors sensitive to $Rg_3$. We further characterized $Rg_3$ regulation of L247T receptors. We found that $Rg_3$ inhibition of mutant ${\alpha}7$ nAChR channel currents was reversible and concentration-dependent. $Rg_3$ inhibition was strongly voltage-dependent and noncompetitive manner. These results indicate that the interaction between $Rg_3$ and mutant receptors might differ from its interaction with the wild-type receptor. To identify differences in $Rg_3$ interactions between wild-type and L247T receptors, we utilized docked modeling. This modeling revealed that $Rg_3$ forms hydrogen bonds with amino acids, such as Ser240 of subunit I and Thr244 of subunit II and V at the channel pore, whereas $Rg_3$ localizes at the interface of the two wild-type receptor subunits. These results indicate that mutation of Leu247 to Thr247 induces conformational changes in the wild-type receptor and provides a binding pocket for $Rg_3$ at the channel pore.
This study was conducted to investigate the changes in saponin content and antioxidant activity of crude ginseng and extruded ginseng by using different solvent extraction methods. Each of the fractions was first extracted by 80% ethanol followed by ether treatment to remove the lipid components. Water soluble components were separated by ethylacetate and water saturated butanol. Four fraction, including 80% ethanol, ethylacetate, butanol and water were obtained from crude and extruded ginsengs to analyze saponin content and antioxidant activity. Saponin content and antioxidant capacity of each of the four fractions were measured by LC/MS analysis and ORAC(Oxygen Radical Absorbance Capacity) assay, respectively. It was found that a major portion of saponin was present in ethyl acetate and water saturated butanol fractions. When extracted by 80% ethanol, ginsenoside Rb1 and Rg1 were mostly found in crude ginseng, while ginsenoside Re and Rb1 were detected in extruded ginseng. Even though Rh1 and Rg3 were found in a very small quantity in crude ginseng, there was a significant quantity of both in extruded ginseng when extracted by 80% ethanol. Similar tendency was also observed in extruded ginseng fraction when extracted with ethyl acetate and butanol. In crude ginseng, the level of Rg1 was the highest among other ginsenosides upon extraction by ethyl acetate, while Rh1 and Rg3 were predominantly found by employing similar solvent extraction in the extruded ginseng. Also, Rg1, Re and Rb1 were also found in the extruded ginseng with small quantity. Rg1, Re and Rb1 were found in crude ginseng by butanol extraction, while Rb1 and Re were extracted from the extruded ginseng. Overall, there was no difference in the saponin content between crude ginseng and extruded ginseng when extracted by butanol and water, but twice as much of saponin was obtained by 80% ethanol extraction and 6 times more saponin were obtained in ethyl acetate fraction in the extruded ginseng. Antioxidant capacity of crude ginseng as determined by ORAC assay was higher in 80% ethanol(high in many different kinds of biological compounds) and water saturated butanol(high in polar saponin) fractions than the ethyl acetate and water fractions. No difference in antioxidant capacity was observed between crude and extruded ginseng. However, antioxidant capacity of ethyl acetate and water fractions in extruded ginseng was significantly higher than crude ginseng($P$ >0.05). All the fractions in both, crude and extruded ginseng possessed antioxidant capacity and even water fractions that contained almost no saponin had some antioxidant capacity. While determining correlation coefficient between fractions in extruded ginseng by Pearson correlation, it was observed that 80% ethanol fraction was in correlation with ethyl acetate($P$ >0.01) and ethanol($P$ >0.001) and in the case of ethylacetate, correlation was observed only with butanol fraction($P$ >0.05).
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