ABSTRACT
Ginsenoside Ro decreased measures of inflammation, aging, oxidants and thrombus formation in a previous study. To measure ginsenoside Ro content in red ginseng from different years, an optimized extraction method was developed to determine ginsenoside Rg1, Re, Rb1 and Ro content by HPLC in 43 batches of red ginseng from different origins, growing years and manufacturers. The results indicate that the best extraction method was to ultrasonify a 1 g sample in 70% methanol for 50 min. The total running time of the optimized gradient was 50 min using a C18 core-shell column and was half the time described in the Chinese Pharmacopoeia, 2015 edition. The separation resolution of all of targeted compounds was greater than 1.6. The peak shape of ginsenoside Ro was optimal when the mobile phase consisted of acetonitrile and water with 0.1% phosphoric acid. The content of ginsenoside Ro was in the range of 0.11% to 0.43%, and the average content was 0.26%, which was higher than that of ginsenoside Rg1 and Re. The ratio of ginsenoside Ro and Re as a threshold could be used to discriminate red ginseng from different growing years; in addition, 100%, 94.4% and 46.6% of red ginseng from six, five and four years exceeded the threshold of 1.3. Our optimized analytical method for characterization of red ginseng is convenient and shortens the assay time.
ABSTRACT
Objective To systematically investigate the chemical constituents of the roots of Panaxginseng. Methods Mild cold-soaked extraction by 70% aqueous ethanol, successive solvent extraction by ethyl acetate and n-butanol, column chromatography by D101 macroporous absorption resin and reversed-phase silica gel, and semi-preparative HPLC, were used for compounds isolation and purification, while high-resolution mass spectrometry, 1D and 2D NMR data were analyzed for compounds identification. Results A new oleanolic acid tetraglycoside (1) and 19 known ginsenosides (2-20) were isolated and identified. Compound 1 was identified as oleanolic acid 3-O-[β-D-glucopyranosyl-(1→2)-β-D-glucuronopyranosyl]-28-O-β-D-glucopyranosyl-(1→6)-β-D-glucopyranoside, named ginsenoside Ro1 (1). The 19 known ginsenosides were notoginsenoside FP1 (2), ginsenoside Re3 (3), notoginsenoside Rt (4), 20-O-glucosyl ginsenoside Rf (5), ginsenoside Re2 (6), ginsenoside Rg2 (7), ginsenoside Ra2 (8), ginsenoside Rb1 (9), ginsenoside Rc (10), ginsenoside Ra1 (11), malonylginsenoside Rb1 (12), malonylfloralginsenoside Rd5 (13), malonylginsenoside Rc (14), ginsenoside Ro (15), ginsenoside Rd (16), ginsenoside F2 (17), 20(R)-ginsenoside Rh2 (18), ginsenoside F3 (19) and ginsenoside F1 (20). Conclusion Compound1is a new compound. Compound 2, notoginsenoside FP1, is isolated from this plant for the first time.
ABSTRACT
Objective:To investigate the promoting effect of Ginsenoside Ro on the differentiation of THP-1-derived dendritic cells (DCs) induced by GM-CSF and IL-4.Methods: Sensitive leukemia-derived DC cell line was screened first.Then,the selected sensitive cell line THP-1 was stimulated to differentiate into DCs by cytokines (GM-CSF and IL-4) and small(5 μmol/L),middle(10 μmol/L),and large (20 μmol/L) dose of Ginsenoside Ro respectively.The expressions of CD1a,MHCⅡ and CD86 of leukemia-derived DCs were detected by flow cytometry.In addition,the transcription levels of CD1a,CD86 and MHCⅡ of leukemia-derived DCs were detected by RT-PCR.ELISA was used to measure the protein levels of TNF-α and IL-6 in the culture supernatant.Results: THP-1 was the sensitive leukemia cell line which could be induced to differentiate into DCs by cytokines.Compared with cytokine stimulation alone,the expression of CD1a,MHCⅡ and CD86 in leukemia-derived cells was significantly increased after the stimulation of Ginsenoside Ro combined with cytokine(P<0.05).The CD1a,CD86 and MHCⅡ mRNA expression was significantly increased after the treatment of Ginsenoside Ro combined with cytokine(P<0.05).Moreover,the protein levels of TNF-α and IL-6 in culture supernatant were significantly increased (P<0.05) after the stimulation of Ginsenoside Ro in combination with cytokines.Conclusion: Ginsenoside Ro can significantly promote the differentiation of leukemia-derived DCs.
ABSTRACT
This study investigated effects of Ginsenoside Ro (Ro) on interleukin-1β (IL-1β)-induced apoptosis and inflammation in rat chondrocytes. The rat chondrocytes were co-treated with IL-1β (10 ng·kg(-1)) and Ro (50, 100 and 200 μmol·L(-1)) for 48 h. Chondrocytes viability was detected by the MTT assay and Annexin V-FITC/PI dual staining assay. Caspase 3 activity was measured by using caspase 3 colorimetric assay kit. Apoptosis related proteins Bax, Bad, Bcl-xL, PCNA, p53 and phospho-p53, along with inflammation related protein MMP 3, MMP 9 and COX-2, and the expression of phospho-NF-κB p65 were assayed by western blotting analyses. Ro could improve IL-1β-induced chondrocytes viability. Ro could suppress IL-1β-induced apoptosis by inhibiting levels of Bax and Bad, decreasing p53 phosphorylation and promoting the expression of Bcl-xL and PCNA. Ro inhibited caspase 3 activity. IL-1β-induced inflammation and matrix degration were also alleviated by Ro with down-regulating the expression of MMP 3, MMP 9 and COX-2. Moreover, Ro inhibited NF-κB p65 phosphorylation induced by IL-1β. In conclusion, these results suggested Ro exerted anti-apoptosis and anti-inflammation in IL-1β-induced rat chondrocytes, which might be related to NF-κB signal pathway. Therefore, we propose that Ro might be a potential novel drug for the treatment of osteoarthritis.
Subject(s)
Animals , Rats , Anti-Inflammatory Agents , Pharmacology , Apoptosis , Apoptosis Regulatory Proteins , Metabolism , Caspase 3 , Metabolism , Cell Survival , Chondrocytes , Cell Biology , Cyclooxygenase 2 , Metabolism , Down-Regulation , Drug Evaluation, Preclinical , Ginsenosides , Pharmacology , Inflammation , Drug Therapy , Interleukin-1beta , Pharmacology , Matrix Metalloproteinase 3 , Metabolism , Matrix Metalloproteinase 9 , NF-kappa B , Metabolism , Phosphorylation , Rats, Sprague-Dawley , Signal TransductionABSTRACT
Aim To study the effects of ginsenoside-Ro on cell proliferation and cytokine production in murine splenocytes. Methods The effect of ginsenoside-Ro on murine splenocytes proliferation was studied using [3 H] thymidine incorporation assay. Effects of ginsenoside-Ro on the production of cytokines interleukin-2 (IL-2), interferon-γ (IFN-γ) and interleukin-4 (IL-4) from murine splenocytes were detected by ELISA method. Effects of ginsenoside-Ro on mRNA level of Th1 cytokine IFN-γ and Th2cytokine IL-4 were evaluated by reverse transcription polymerase chain reaction (RT-PCR) analysis.Results Ginsenoside-Ro showed no mitogenic effect on unstimulated murine splenocytes. It enhanced the proliferation of Con A-induced murine splenocytes and the production of IL-2 at concentrations of 1 - 10decreased the production and expression of Th1 cytokine IFN-γ in Con A-induced murine splenocytes at regulating the production and expression of Th1/Th2 cytokines in murine splenocytes.