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Chikusetsusaponin IVa (CsIVa) is a natural active monomer of triterpene saponins in the Chinese herbal medicine of Panax japonicus, which has anti-inflammatory, anti-tumor and other effects. However, its function and mechanism in triple negative breast cancer (TNBC) remain unclear. This study investigated the inhibitory effect and mechanisms of CsIVa on the proliferation of triple negative breast cancer cell line MDA-MB-231. In this study, we found that CsIVa could significantly inhibit the proliferation of MDA-MB-231 cells and eliminate its potential toxic effect on normal breast cells (MCF-10A). The transcriptome sequencing results showed that the inhibition of proliferation of MDA-MB-231 cells by CsIVa was closely related to cell cycle and the pathway regulating cell cycle. Further studies confirmed that CsIVa blocked the cell cycle in G2/M phase by down-regulating the expression of cyclin dependent kinase 1 (CDK1), cyclin B1 and up-regulating the expression of cyclin dependent kinase inhibitor 1A (p21). Moreover, CsIVa can block cell cycle through inhibiting PI3K/AKT signal pathway. In conclusion, CsIVa regulates the expression of cell cycle related proteins (p21, CDK1, cyclin B1) via inhibiting the activity of PI3K/AKT signaling pathway, blocks TNBC cell cycle, and thus exerts its anti-tumor activity.
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Objective To establish and identify HPLC fingerprints of Panacis Japonici Rhizoma from different regions, and provide a scientific method for quality its control and standardization. Methods The fingerprints of Panacis Japonici Rhizoma were established based on HPLC method. The similarity evaluation, cluster analysis (CA), and principal component analysis (PCA) were applied for the experimental data, in order to find out the similarities and differences among the 10 batches of Panacis Japonici Rhizoma from different regions. Meanwhile, the qualitatively and quantitatively analyze were adopted in the common peaks of Panacis Japonici Rhizoma by total statistical moment method. Results The HPLC fingerprint with 16 common peaks was established, for the common chemical components, six were identified as ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, chikusetsusaponin V, chikusetsusaponin IV, and chikusetsusaponin IVa, respectively. Among them, the contents of chikusetsusaponin V and IV in different batches of Panacis JaponiciRhizoma samples showed little differences. The similarities of samples from various regions except S7 were over 0.90. The samples were classified into three categories by CA and PCA methods. The parameters of total statistical moment for five species of Panax genus showed significantly differences. Conclusion The developed HPLC fingerprint of Panacis Japonici Rhizoma combined with chemical pattern recognition can provide a scientific instruction for the quality control of Panacis Japonici Rhizoma and the differentiation of five herb medicines from Panax genus.
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Objective: To establish a simple and effective extraction method for the preparation of total saponins of Panax japonicas (TSPJ). Methods: Combination of macroporous adsorption and ion exchange resin chromatography was adopted in the present study. For quality evaluation, chikusetsusaponin IVa was used as reference, and vanillin-perchloric acid was applied as chromogenic reagent to determine total saponin content at 545 nm. Results: X-5 macrophous resin offered better adsorption and desorption capacities for TSPJ than other macrophous resins. The optimum purification process was confirmed as follows: The sample solution concentration was 0.2 mg/L; The sample volume was 10 g/g, and eluting with 5 mL of 70% aqueous ethanol solutions on 1 g wet macrophous resin column. Followed this step, decoloring of TSPJ was studied and the decoloring capacity of two different types of ion exchange resins was evaluated. The result showed that 732-type cation exchange resin was the better resin for decolorization of the TSPJ. The total saponin products with higher purity and quality were obtained, with the mass fraction more than 85.0%, and the transfer rate of TSPJ was more than 70.0%. Conclusion: The results show that the total saponins can be separated and purified effectively from P. japonicus. The preparation method is simple, effective, and efficient for large-scale preparation of TSPJ.
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Objective: A high-performance liquid chromatography-tandem mass spectrometric method for the simultaneous determination of 15 ginsenoside compounds from Panacis Majoris Rhizoma (PMR) was developed. Methods: A Waters Sunfire ™ C18 column (150 mm × 4.6 mm, 5 μm) was used for the separation. The mobile phase consisted of A (H2O + 0.05% HCOOH) and B (CH3CN + 0.05% HCOOH) using a gradient elution. For the quantification of ginsenosides, the multiple reaction monitoring (MRM) mode of the mass spectrometer was applied and the declustering potential (DP), collision energy (CE), and collision cell exit potential (CXP) were optimized to perform automatic on-line MS/MS experiments during the chromatographic separation. Results: By using the optimized method, the linearity range of 15 analytes was 0.000 9 to 2 952.592 3 μg/mL with more than 0.999 determination coefficient (r) of linear regressions, the detection limits of the 15 ginsenosides ranged from 0.003 to 626.554 ng/mL, the limits of quantitation ranged from 0.075 to 1 762.150 ng/mL, the recoveries of 15 ginsenosides in the samples were 98.15%-101.12% with relative standard deviation (RSD) that ranged from 0.82% to 2.15%. Conclusion: The proposed LC-MS/MS method is accurate and reproducible in accordance with TCM guidelines, showing high sensitivity, rapidness, and recovery. This method allows the assessment of various ginsenosides in a single analytical run providing an innovative tool to control Panacis Majoris Rhizoma materials quantification.
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Objective: To investigate the chemical constituents in the dried leaves of Sabia fasciculata. Methods: Column chromatography such as silica gel, Sephadex LH-20, and preparative HPLC were used to isolate and purify the compounds. Spectroscopic methods like MS, 1H-NMR and 13C-NMR, and physical constants were used to elucidate their structures. Results: Fifteen compounds were isolated from 95% ethanol extracts of S. fasciculata, including seven pentacyclic triterpenoids, such as methyl-3-oxo-olean-12-ene-28-oate (1), betulin (2), 3-oxo-olean-Δ11,13(18)-diene (3), oleanolic acid (4), imberic acid (5), pseudo- ginsenoside RP1 (6), and chikusetsusaponin IVa (7); three flavonoids, such as quercetin (8), rutin (9), and mutabiloside (10); three alkaloids, such as fuseine (11), N-p-feruloyltyramine (12), and N-trans-coumaroyl tyramine (13); two steroids, such as β-sitosterol (14) and β-daucosterol (15). Conclusion: All the compounds are isolated from the plant for the first time. Compounds 1, 5-7, and 10 are isolated from the plants of Sabia Colelbr. for the first time.