Systems pharmacology dissection of action mechanisms for herbs in osteoporosis treatment / 中草药·英文版

Objective: Osteoporosis has become the biggest cause of non-fatal health issue. Currently, the limitations of traditional anti-osteoporosis drugs such as long-term ill-effects and drug resistance, have raised concerns toward complementary and alternative therapies, particularly herbal medicines and their natural active compounds. Thus, this study aimed to provide an integrative analysis of active chemicals, drug targets and interacting pathways of the herbs for osteoporosis treatment. Methods: Here, we introduced a systematic pharmacology model, combining the absorption, distribution, metabolism, and excretion (ADME) screening model, drug targeting and network pharmacology, to probe into the therapeutic mechanisms of herbs in osteoporosis. Results: We obtained 86 natural compounds with favorable pharmacokinetic profiles and their 58 targets from seven osteoporosis-related herbs. Network analysis revealed that they probably synergistically work through multiple mechanisms, such as suppressing inflammatory response, maintaining bone metabolism or improving organism immunity, to benefit patients with osteoporosis. Furthermore, experimental results showed that all the five compounds (calycosin, asperosaponin VI, hederagenin, betulinic acid and luteolin) enhanced osteoblast proliferation and differentiation in vitro, which corroborated the validity of this system pharmacology approach. Notably, gentisin and aureusidin among the identified compounds were first predicted to be associated with osteoporosis. Conclusion: Herbs and their natural compounds, being characterized as the classical combination therapies, might be engaged in multiple mechanisms to coordinately improve the osteoporosis symptoms. This work may contribute to offer novel strategies and clues for the therapy and drug discovery of osteoporosis and other complex diseases.

Methyl jasmonate stimulates the synthesis and accumulation of asperosaponin VI in the roots of Dipsacus asper / 药学学报

We investigated the effect of methyl jasmonate (MeJA) on the content of asperosaponin VI and the expression of genes involved in its synthesis. Dipsacus aspero seedlings were treated with MeJA at different concentrations of 50, 100, 150, 200 and 300 μmol·L-1, and leaves and roots were sampled following treatment for 1, 3 and 5 days. The content of asperosaponin VI and superoxide anion in the roots, malondialdehyde (MDA) content in leaves and superoxide dismutase were determined. The results show that 150 μmol·L-1 MeJA significantly increased the accumulation of asperosaponin VI in roots. The content of asperosaponin VI was greatest after treatment for 3 days, and was 2.16 times higher than the control. After MeJA treatment, SOD activity decreased and MDA content increased in leaves. Moreover, superoxide anion content in roots increased. The expression of squalene epoxidase (DaSE1) and geranyl diphosphate synthase (DaGPS), key enzymes in the synthesis of asperosaponin VI, were up-regulated compared with the control group. These results indicate that an optimal concentration of 150 μmol·L-1 MeJA increases the accumulation of asperosaponin VI by up-regulating the expression of key enzymes involved in the synthesis of asperosaponin VI, which facilitates resistance to adversity stress stimulated by MeJA.

Analysis of components of crude and sweated Dipsaci Radix by UPLC-Triple-TOF/MS / 中草药

Objective: To establish a quick method of ultra-performance liquid chromatography-quadrupole time-of-fight mass spectrometry (UPLC-Triple-TOF/MS) for the analysis of components of crude and sweated Dipsaci Radix. Methods: The separation was performed on the chromatographic column of Agilent Eclipse XDB-C18 (250 mm × 4.6 mm, 5.0 μm), and the mobile phase was 0.1% formic acid solution-methanol, with a gradient elution at a flow rate of 0.8 mL/min, the detection wavelength was 215 nm, the column temperature was 25 ℃. UPLC-Triple-TOF 5600+ time of flight liquid and mass spectrometer was used for mass spectrometry. Electrospray ion source negative ion mode was adopted, and the scanning range was m/z 100-1 500. The components of crude and sweated Dipsaci Radix were quickly identified according to the information obtained by high-resolution mass spectrometry combined with secondary mass spectrometry. Results: Fifty-two common components were identified or tentatively characterized based on the retention time and MS spectra. They were triterpenoid saponins, iridoids, phenolic acids etc. The crack rules of primary components were also analyzed. And comparing the components of crude and sweated Dipsaci Radix, it showed that the content of 20 components such as loganin acid, chlorogenic acid, loganin, isochlorogenic acid A, and asperosaponin VI was decreased after sweating, and caffeic acid, isochlorogenic acid, isochlorogenic acid C, and triplostoside A was increased. Conclusion: The types of components of crude and sweated Dipsaci Radix are identical, but there are differences in the content of the components. The content of the components of crude are higher than the sweated Dipsaci Radix. UPLC-Triple-TOF-MS technology was used to analyze the influence of “sweating” on the chemical composition of the Dipsaci Radix, so as to provide a theoretical basis for the study of the chemical constituents of sweated Dipsaci Radix and further research on the origin processing of Dipsaci Radix.

Spectral-effect relationship between HPLC fingerprint and cell proliferation and differentiation of crude and sweated Dipsaci Radix / 中草药

Objective: To study the spectrum-effect relationships of fingerprints of sweated and crude Dipsaci Radix on cell proliferation and differentiation, and find out the material basis of efficacy before and after sweating in order to provide the basis for the impact of the efficacy. Methods: The DAD detector was used to establish HPLC fingerprints of sweated and crude Dipsaci Radix, and the relationship between the spectrum and efficiency was established by gray relational analysis. Results: The chemical composition of peaks 14 and 4 were highly correlated with the proliferation and differentiation of osteoblasts and the proliferation of MG-63 cells. The correlations were all above 0.7. The three pharmacophore indicators associated much with the characteristic peaks 14, 4, 6, 16, 13, 11, and 5. Combined with the previous analysis, these peaks represented respectively asperosaponin VI, chlorogenic acid, loganin, asperosaponin IV isomers, dipsacoside X, chlorogenic acid C, and caffeic acid. Conclusion: Asperosaponin VI, chlorogenic acid, loganin, asperosaponin IV isomers, dipsacoside X, chlorogenic acid C, and caffeic acid may be the main material basis for the effect of cell proliferation and differentiation, thus affecting its efficacy.

Correlation of powder particle size and in vitro dissolution rate in Xuejie Sanqi Jiegu Paste based on multi-components determination / 中草药

Objective To investigate the relationship between powder particle size and dissolution of Xuejie Sanqi Jiegu Paste (XSJP) in vitro. Methods The particle size of three kinds of XSJP was measured by laser particle size instrument. The hydroxysafflor yellow A, asperosaponin VI, ginsenoside Rg1, ginsenoside Rb1, notoginsenoside R1, dracorhodin were taken as indexes, and the feature of dissolution for the above three kinds of XSJP was examined by grouting method. The correlation between the powders in different particle sizes and their corresponding dissolution was analyzed by software SPSS 17.0. Results The particle size of the three kinds of powders was correlated with the dissolution of index components. The powders in different particle sizes were negatively correlated with the relative cumulative dissolution of hydroxysafflor yellow A, asperosaponin VI, ginsenoside Rg1, ginsenoside Rb1 and notoginsenoside R1. The cumulative dissolution of superfine powder was respectively increased by 6.8%, 16.6%, 6.6%, 6.7%, and 5.4%, while the dracorhodin had no direct correlation. Conclusion There is a correlation between the particle size and dissolution of Chinese medicine with fibre which contains cellular structure.

A quantitative method using one marker for assay of multi-components in Xianling Gubao Capsule / 中草药

Objective: To investigate the positioning based on the relative retention time of fingerprinting and to establish a new quality evaluation method for traditional Chinese medicine preparations, using one chemical reference substance to calcutate multi- components simultaneously. Methods: Employed icariin as the maker component, icariin relative correction factors (RCF) of epimedin C to icariin, asperosaponin VI to icariin, psoralen to icariin and angelicin to icariin were ealeatated in the chromatographic conditions for determination of the four components in Xianling Gubao Capsule (XGC). The contents of icariin were determined by external standard method, and those of epimedin C, asperosaponin VI, psoralen and angelicin were calculated by icariin and their RCF. The accuracy of the new method was evaluated by comparing the relative retention times and calculating the content which using different brands columns for determination. Results: The analysis methods were established,the linearity was good when sample volume was in the range of at 8.2—328.0 ng for icariine(r = 0.999 5), 0.055 6—2.224 μg for epimedin C (r = 0.999 6), 0.144 1—5.764 μg for asperosaponin VI (r = 0.999 6), 5.4—215.2 ng (r = 0.998 0) for psoralen, 6.6—265.6 ng (r = 0.998 5) for angelicin. The average recoveries of asperosaponin VI, psoralen and psoralen were 97.59%, 98.58%, 98.11%, 97.86%, 98.22%, respectively. The RSDs of recovery were all less than 2.0%; There has been no significant difference between the calculated contents and the determined contents, according to the angle cosine value. Conclusion: The method can control the components without providing epimedin C, asperosaponin VI, psoralen, and angelicin reference. The method is not only save reference substance and medicine resources, but also suitable quality evaluation pattern for TCM preparation. This new method made fingerprinting more meaningful in TCM quality control.

Simultaneous determination of 12 chemical components in Tiaojing Pills by RP-HPLC under variable wavelengths / 中草药

Objective: To establish a RP-HPLC method to determine the contents of paeoniflorin, acteoside, ferulic acid, leonurine hydrochloride, hesperidin, paeonol, baicalin, asperosaponin VI, limonin, atractylenolide I, atractylenolide III, and pachymic acid in Tiaojing Pills. Methods: The determination was performed on a Venusil MP-C18 column (250 mm × 4.6 mm, 5.0 μm) with ethanol-acetonitrile (40∶60, A) and 0.2% phosphoric acid (B) as mobile phases for gradient elution, at the flow rate of 0.8 mL/min; The column temperature was 45 ℃. Results: The nine components were well separated and showed good linearity, such as paeoniflorin 0.5-50.0 mg/L (r = 0.999 5), acteoside 0.1-10.0 mg/L (r = 0.999 1), ferulic acid 0.2-20.0 mg/L (r = 0.999 2), leonurine hydrochloride 0.3-30.0 mg/L (r = 0.999 3), hesperidin 4.0-400.0 mg/L (r = 0.999 8), paeonol 0.2-20.0 mg/L (r = 0.999 1), baicalin 0.6-60.0 mg/L (r = 0.999 4), asperosaponin VI 1.5-150.0 mg/L (r = 0.999 8), limonin 7.0-700. 0 mg/L (r = 0.999 9), atractylenolide I 0.5-50. 0 mg/L (r = 0.999 3), atractylenolide III 0.5-50. 0 mg/L (r = 0.999 4), and pachymic acid 1.0-100. 0 mg/L (r = 0.999 6). The precision was good, RSD ≤ 0.97%, the repeatability was good in terms of RSD ≤ 1.25% and the recovery rate was 98.5%-103.5% (RSD ≤ 1.24%). Test solution was stable at room temperature within 24 h. The contents of twelve batches of the paeoniflorin, acteoside, ferulic acid, leonurine hydrochloride, hesperidin, paeonol, baicalin, asperosaponin VI, limonin, atractylenolide I, atractylenolide III and pachymic acid were 4.328-4.688, 0.033-0.054, 0.073-0.091, 0.177-0.199, 0.243-0.283, 0.043-0.069, 1.144-1.173, 0.037-0.061, 0.094-0.126, 0.127-0.157, 0.155-0.179, and 0.285-0.327 mg/g, respectively. Conclusion: The method is rapid and has high sensitivity, high accuracy, and good specificity, It can be applied to the quality control of Tiaojing Pills.

Determination of asperosaponin VI, psoralen, and angelicin in Xianling Gubao Capsule by HPLC / 中草药

Objective: To determine asperosaponin VI, psoralen, and angelicin in Xianling Gubao Capsules (XGC) via multi- wavelength HPLC method. Methods: Separation was carried out on Welch Ultimate® XB-C18 column. The mobile phase was acetonitrile-water system and a linear gradient elution was used. The column temperature was 30℃. The detection wavelength for asperosaponin VI was set at 212 nm, those for psoralen and angelicin were set at 246 nm. Results: Three components reached baseline separation, the linearity was good when sample volumes were in the ranges of 144.1-5 764.0 for asperosaponin VI (r = 0.999 6), 5.4-215.2 (r = 0.998 0) for psoralen, and 6.6-265.6 ng (r = 0.998 5) for angelicin. The average recoveries of asperosaponin VI, psoralen, and psoralen were 98.11%, 97.86%, and 98.22%, respectively. The RSDs of recoveries were all less than 2.0%. Conclusion: The method is simple and accurate and has good separation, with high sensitivity and good efficiency for the determination of more-index components in XGC.

Isolation, structure characterization and quantification of related impurities in asperosaponin VI / 中国天然药物

AIM@#To study the related impurities in asperosaponin VI bulk drug and to develop a high performance liquid chromatography (HPLC) method for the determination of asperosaponin VI and its related impurities.@*METHODS@#The related impurities were detected in asperosaponin VI bulk drug by a newly developed HPLC method, obtained by ODS column chromatography and semi-preparative HPLC methods, and the structures were elucidated by TOF-MS, IR, and NMR techniques. The HPLC method was validated according to ICH guidelines for asperosaponin VI and its related impurities.@*RESULTS@#Seven related impurities (Imp 1-7) were isolated from asperosaponin VI bulk drug. Impurity 3 was found to be a mixture of two epimers, and was first reported in the paper. The validation results showed good sensitivity, specificity, linearity (r(2) ≥ 0.997 9), precision (RSD < 5.0%), accuracy (recoveries in the range of 94.61%-106.51%) and robustness.@*CONCLUSION@#The developed HPLC method is suitable for the quality control of asperosaponin VI bulk drug.