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1.
Chinese Journal of Natural Medicines (English Ed.) ; (6): 494-505, 2022.
Article in English | WPRIM | ID: wpr-939914

ABSTRACT

Impaired immunomodulatory capacity and oxidative stress are the key factors limiting the effectiveness of mesenchymal stem cell transplantation therapy. The present study was aimed to investigate the effects of jujuboside A (JuA) on the protective effect and immunomodulatory capacity of human umbilical cord mesenchymal stem cells (hUC-MSCs). Hydrogen peroxide was used to establish an oxidative damage model of hUC-MSCs, while PBMCs isolated from rats were used to evaluate the effect of JuA pre-treatment on the immunomodulatory capacity of hUC-MSCs. Furthermore, Hoechst 33258 staining, lactate dehydrogenase test, measurement of malondialdehyde, Western blot, high-performance liquid chromatography; and flow cytometry were performed. Our results indicated that JuA (25 μmol·L-1) promoted the proliferation of hUC-MSCs, but did not affect the differentiating capability of these cells. JuA pre-treatment inhibited apoptosis, prevented oxidative damage, and up-regulated the protein expression of nuclear factor-erythroid factor 2-related factor 2 and heme oxygenase 1 in hUC-MSCs in which oxidative stress was induced with H2O2. In addition, JuA pre-treatment enhanced the inhibitory effect of hUC-MSCs against abnormally activated PBMCs, which was related to stimulation of the expression and activity of indoleamine 2,3-dioxygenase. In conclusion, our results demonstrate that JuA pre-treatment can enhance the survival and immunomodulatory ability through pathways related to oxidative stress, providing a new option for the improvement of hUC-MSCs in the clinical setting.


Subject(s)
Animals , Humans , Rats , Cell Differentiation , Hydrogen Peroxide/metabolism , Mesenchymal Stem Cells , Oxidative Stress , Saponins , Umbilical Cord/metabolism
2.
Chinese Traditional and Herbal Drugs ; (24): 2712-2717, 2019.
Article in Chinese | WPRIM | ID: wpr-851104

ABSTRACT

Objective: To study and compare the content of two flavonoids and two saponins in Ziziphi Spinosae Semen from seven major producing areas in China, and to provide high quality sources of medicinal materials. Methods: Investigation and sample collection of seven major production areas were carried out in 2016 and 2017 and a total of 74 samples were collected. UPLC-UV-ELSD liquid phase method was used to determine the content of two flavones, spironol and 6’’’-ferulinyl spironol, as well as two saponins jujuboside A and jujuboside B. Results: In 2016 and 2017, the content of spinolin in the samples was 0.052%-0.102% and 0.049%-0.144%, respectively. The content of 6’’’-ferulinyl spironol was 0.021%-0.072% and 0.026%-0.088%, respectively. The content of jujuboside A was 0.016%-0.061% and 0.033%-0.054%, respectively. The content of jujuboside B was 0.008%-0.046% and 0.005%-0.046%, respectively. Conclusion: By correlation analysis, there was no significant difference in the content of each component among the seven major production areas. The producing areas with high spinosin content are Linfen, Shanxi (including Daning and Jixian), Shexian, Hebei, Qian’ an, Hebei, Qingyang, Gansu, Meixian, Shaanxi, Heyang, Shaanxi, Jianchang, Liaoning, and Jixian, Tianjin. The producing areas with high content of jujuboside A are most places of Shandong province, Neiqiu, Hebei and Weinan, Shaanxi. Considering the content of two kinds of components, Shandong Province is generally high and has little change. The correlation analysis also showed that the content of two flavonoids had good correlation, but the content of two saponins had poor correlation.

3.
Chinese Traditional and Herbal Drugs ; (24): 4634-4642, 2019.
Article in Chinese | WPRIM | ID: wpr-850811

ABSTRACT

Objective: The aim of this work was to investigate the quality markers (Q-markers) of Ziziphi Spinosae Semen (ZSS) based on biotransformation by human intestinal microbiota. Methods: In this study, in vitro biotransformation of ZSS aqueous extract by normal human intestinal microbiota was analyzed using UPLC-Q-Orbitrap-MS. Furthermore, the time course of the biotransformation was studied to probe into the biotransformation mechanism of compounds in ZSS by human intestinal flora. The change rules of flavonoids, saponins and alkaloids in the incubation solution at different time points were plotted based on the percentage of peak area of compounds. Results: A total of 31 original ingredients and four metabolites were characterized in transformed ZSS aqueous extract by human intestinal microbiota. No obvious degradation was observed for benzylisoquinoline alkaloids within 24 h. As far as flavones concerned, a wide range of metabolic reactions as well as significant reaction were shown. Meanwhile, these flavonoids were completely degraded during 24 h. In addition, both jujuboside A and jujuboside B were metabolized to their saponins by deglycosylation reactions. Thus, coclaurine, zizyphusine, kaempferol-3-O-rutinoside, spinosin, vicenin II, jujuboside A, and jujuboside B were referred as prospective Q-markers. Conclusion: The results indicated that the chemical compounds in ZSS were obviously affected by transformation. Intestinal transformation studies play an important role for the elucidation of therapeutic material basis of ZSS and it should be taken into account during the process of the investigation of Q-marker.

4.
China Pharmacy ; (12): 2802-2807, 2019.
Article in Chinese | WPRIM | ID: wpr-817524

ABSTRACT

OBJECTIVE: To establish a method for simultaneous determination of spinosin and jujuboside A in the seads of Ziziphus jujuba, and to investigate its quality grading standard. METHODS: HPLC-ELSD method was adopted. The separation was carried out on Inertsil ODS-SP column with mobile phase consisted of acetonitrile-water (gradient elution) at the flow rate of 1.0 mL/min. The column temperature was 30 ℃, the temperature of drift tube was 90 ℃, the flow of carrier gas was 2.9 L/min and injection volume was 20 μL. The thickness, width, length and 100-grain quality of the medicinal materials were used as indicators to investigate the appearance traits. SPSS 22.0 software was used to analyze the correlation of the contents of spinosin and jujuboside A, its appearance traits with the quality constant of TCM, and establish a quality classification standard for the seads of Z. jujuba. RESULTS: The linear range of spinosin and jujuboside A were 1.03-6.18 μg/mL (r=0.999 7), 1.05-6.30 μg/mL (r=0.999 8); the limits of quantitation were 0.171, 0.174 μg/mL, respectively; the limits of detection were 0.052, 0.053 μg/mL, respectively. RSDs of precision, stability and reproducibility tests were all lower 2%. The recoveries were 99.01%-102.97% (RSD=1.39%, n=6), 97.94%-101.03% (RSD=1.13%, n=6), respectively. Correlation analysis results showed that the length, width, 100-grain quality spinosin content and jujuboside A content of the medicinal materials were positively correlated with the quality constant of TCM. The results of quality classification for 30 batches of medicinal materials showed that S1-S4 and S7-S12 were first-class products; S5, S6, S13-S17 and S20-S30 were second-class products; S18 and S19 were third-class products. CONCLUSIONS: Established content determination method is simple, precision, accurate and stable, and can be used for simultaneous determination of spinosin and jujuboside A in the seads of Z. jujuba. Established quality grading standard of the seads of Z. jujuba can be used to evaluate the quality.

5.
Chinese Traditional and Herbal Drugs ; (24): 4520-4527, 2018.
Article in Chinese | WPRIM | ID: wpr-851652

ABSTRACT

Objective To establish a quality evaluation system for slice decoction of Ziziphi Spinosae Semen (SZR). Methods Ten batches of SZR slice were collected and prepared into decoction, and then the decoctions of SZR were prepared as freeze-dried powders. The extraction rate and the content of the index components were used as indicators to optimize the extraction process. Multi-dimensional chemical fingerprints were established by HPLC coupled with UV detector and ELSD detector. The similarity, RSD of relative retention time, and RSD of relative peak area were analyzed to comprehensively evaluate the quality of ten batches of SZR slice decoction. A sensitive method based on ultra-HPLC coupled with triple quadrupole mass spectrometry (UPLC-MS/MS) was established and validated for quantitative analysis of the nine compounds in SZR slice decoction, including coclaurine, magnoflorine, vicenin II, spinosin, swertisin, kaempferol-3-O-rutinoside, 6'''-feruloylspinosin, jujuboside A, and jujuboside B. Consequently, the developed methods were successfully applied to the quality assessment of SZR slice decoction. Results The average dry extraction rate of ten batches of SZR slice decoction was 20.77%. Three, four, and two common peaks were identified in the HPLC-UV 227 nm, 335 nm, and HPLC-ELSD fingerprints. The similarity of the fingerprints at different detection wavelengths was in the range of 0.942-0.988, relative retention time and relative peak area RSD% were less than 3%. The method validation results indicated that the methods were simple, specific and reliable. All the compounds showed good linearity (r > 0.999 0) with a relatively wide concentration range, acceptable recovery at 87.40%-110.42%, and RSD% less than 4.12%. The content of nine chemical compounds in ten batches of SZR slice decoction were as follows: 0.25-0.27 mg/g of coclaurine, 3.2-3.6 mg/g of magnoflorine, 0.077-0.084 mg/g of vicenin II, 0.82-0.89 mg/g of spinosin, 0.007 6-0.008 5 mg/g of swertisin, 0.057-0.060 mg/g of kaempferol-3-O-rutinoside, 0.32-0.35 mg/g of 6'''-feruloylspinosin, 0.83-0.91 mg/g of jujuboside A, and 0.095-0.110 mg/g of jujuboside B. Conclusion A stable and reliable quality evaluation method of SZR slice decoction was established, which provides a stable material basis for the follow-up pharmacokinetic and pharmacodynamic experiments and provides a reference for the quality control of the prescription containing SZR at the same time.

6.
Chinese Traditional and Herbal Drugs ; (24): 2002-2006, 2017.
Article in Chinese | WPRIM | ID: wpr-852797

ABSTRACT

Objective: To establish an accelerated solvent extraction (ASE)-charged aerosol detector (CAD) method for simultaneous detection of jujuboside A, jujuboside B, spinosin, and betulinic acid in Zizyphi Spinosae Semen. Methods: The orthogonal design was applied to optimize the extraction parameters of the ASE system. The target compounds were detected by HPLC-CAD with the parameters as follow: Thermo Syncronis C18 (100 mm × 3 mm, 3 μm) column, a gradient elution program with acetonitrile-water as mobile phase at a flow rate of 0.5 mL/min, the column temperature was kept at 40℃. Detector was Corona Ultra CAD with 35℃ of nebulization temperature. Results: Optimization of the ASE parameters with orthogonal design greatly improved the extraction efficiency; All the target compounds could be simultaneously determined in a single run. Good linear relationships (0.998 3-0.999 6) and high relative recoveries were 98.46%-102.02%. Conclusion: The method is rapid, simple, accurate, and thus could be used for the quality control of Zizyphi Spinosae Semen.

7.
Chinese Journal of Pharmacology and Toxicology ; (6): 399-404, 2017.
Article in Chinese | WPRIM | ID: wpr-613832

ABSTRACT

OBJECTIVE To investigate the effect of jujuboside A on glomerular cell apoptosis in diabetic rats, and to explore the possible mechanisms. METHODS SD rats were administered with streptozotocin 100 mg · kg-1 to estabilish the diabetic model. Diabetic SD rats received jujuboside A 10 and 20 mg · kg-1 daily for 4 weeks by lavage administration, respectively. The level of glycosylated hemoglobin (GHb) in the blood of each group was measured by fructosamine method. The morphological changes in glomerular cells were observed by PAS staining. Glomerular cell apoptosis was determined by TUNEL staining. The protein expression of Bcl-2 and Bax was detected by immunohistochemistry. The protein expression of cleaved caspase 9 and cleaved caspase 3 was detected by Western blotting. Trans?forming growth factor β1 (TGF-β1) mRNA expression was analyzed by qPCR. RESULTS Compared with model group, jujuboside A 10 and 20 mg·kg-1 treatment significantly reduced the level of GHb in blood (mmol · L- 1: 10.9 ± 0.8 vs 17.5 ± 1.5, P<0.05; 7.6 ± 0.5 vs 17.5 ± 1.5, P<0.05), PAS positive score of glomerular cells (26.8 ± 3.2 vs 36.4 ± 3.8, P<0.05; 18.4 ± 2.1 vs 36.4 ± 3.8, P<0.05) and the apoptosis of glomerular cells〔(8.2±0.8)%vs (17.6±1.8)%, P<0.05;(5.1±0.5)%vs (17.6±1.8)%, P<0.05〕. Moreover, Bcl-2 protein expression in kidney tissues was elevated (P<0.05), whereas Bax (P<0.05), cleaved caspase 9 (P<0.05) and cleaved caspase 3 (P<0.05) protein expression and TGF-β1 mRNA (P<0.05) expression were reduced after jujuboside A administration. CONCLUSION Jujuboside A can prevent glomerular cell apoptosis in diabetic rats, which may be associated with the regulation of mitochondrial apoptotic pathways and TGF-β1 expression.

8.
Chinese Pharmaceutical Journal ; (24): 751-756, 2016.
Article in Chinese | WPRIM | ID: wpr-859128

ABSTRACT

OBJECTIVE: To establish an HPLC method for simultaneous determination of ginsenoside Rg1, Re, Rb1 and jujuboside A, B in Renshen Jianpi Pellets (RSJPW). METHODS: The determination was conducted on an Eclipse XDB C18 column (4.6 mm × 250 mm, 5 μm) with the mobile phase of acetonitrile (A)-water (B) gradiently eluted at the following flow rates: 0-90 min, 1.0 mL · min-1; 90-130 min, 1.0-0.5 mL · min-1; 130-140 min, 0.5 mL · min-1. And the column temperature was maintained at 35℃; the detection wavelength was set at 203 nm. RESULTS: The calibration curves of ginsenoside Rg,, Re, Rb, and jujuboside A, B were in good linearity over 0.32-2.24 μg (r=0.9982), 0.16-1.12 μg (r=0.995 3), 0.32-2.24 μg (r=0.9996), 0.16-1.12 μg (r=0.9915), 0.08-0.56 μg (r=0.9996). The corresponding average recovery rates were 97.18%, 97.62%, 98.79%, 98.48%, 94.51%; the standard deviations were 1.10%, 0.98%, 0.34%, 1.09%, 1.88%, respectively. CONCLUSION: The established HPLC method is accurate, reliable and reproducible, which can be used as a reference for the quality control of RSJPW.

9.
China Journal of Chinese Materia Medica ; (24): 1880-1883, 2016.
Article in Chinese | WPRIM | ID: wpr-250473

ABSTRACT

Immunogenic antigen (jujuboside A-BSA) and coating antigen (jujuboside A-OVA) of jujuboside A were synthesized by sodium periodate oxidation method for the first time. Jujuboside A artificial antigen was confirmed by matrix-assisted laser desorption ionization/time-of-flight mass spectrometry (MALDI-TOF-MS). The titer and specificity of the antibody in serum of immunized mice were detected by enzyme-linked immunosorbent assay (ELISA). The corrected relation curve of inhibition rate showed that the antibody against Jujuboside A obtained from immunized mice could bind to jujuboside A and the titer was up to 1∶4 000. The jujuboside A artificial antigen was synthesized, which can be used further to preparation of monoclonal antibody and the pharmacokinetics study of jujuboside A in laboratory animals.

10.
China Pharmacist ; (12): 1073-1076, 2016.
Article in Chinese | WPRIM | ID: wpr-494930

ABSTRACT

Objective:To develop a method of quantitative analysis of multi-components by single marker ( QAMS) for 4 kinds of saponins in spina date seed.Methods:Using jujuboside A as a reference, an HPLC method with a Kromasil C18 column (250 mm × 4.6 mm, 5 μm) was applied, the mobile phase was acetonitrile-0.1% phosphoric acid (30 ∶70), the flow rate was 1.0 ml · min-1 , the detection wavelength was 280 nm, the column temperature was 32℃and the injection volume was 20μl.The relative cor-rection factors among the four kinds of saponins were detected by QAMS .An external standard method was also used to determine the contents of the four saponins ( Wf ) , and the results were compared with those of QAMS ( Ws ) , and then the relative errors were calcu-lated between W f and Ws to evaluate the accuracy of QAMS , and the reproducibility was investigated as well .Results:The established QAMS was used to determine the four kinds of saponins in spina date seed , and totally 14 batches of spina date seed were determined . No significant differences were found out between the calculated values and the determined ones ( RSD<5%) .Conclusion:The vali-dated HPLC method shows the advantages of precision ,reproducibility and reliability .The established QAMS method is feasible , and suitable for the quality control of saponins in spina date seed .

11.
Chinese Traditional and Herbal Drugs ; (24): 751-755, 2015.
Article in Chinese | WPRIM | ID: wpr-854354

ABSTRACT

To analyze and evaluate the quality of Ziziphi Spinosae Semen produced in Tianjin and comparing with Ziziphi Spinosae Semen traded on the market in different places. Twelve kinds of wild Ziziphi Spinosae Semen samples collected from the four represent regions were gathered by systematic combination of regional stratified sampling method and HPLC method was used to determine spinosin, jujuboside A, jujuboside B, and betulinic acid. The contents of spinosin, jujuboside A, jujuboside B, and betulinic acid from wild Ziziphi Spinosae Semen produced in Tianjin are 0.810-1.925, 0.695-1.708, 0.201-0.390, and 0.651-0.789 mg/g; Ziziphi Spinosae Semen produced in Chengde, Hebei province has the highest betulinic acid contents of 1.654 mg/g. The quality of wild Ziziphi Spinosae Semen produced in Tianjin is excellent. What's more, the wild Ziziphi Spinosae Semen produced in Baxian Mountain, Tianjin takes particularly prominent advantages of better quality. Quality of wild Ziziphi Spinosae Semen produced in other three regions is equal to the average level of that traded on the market.

12.
World Science and Technology-Modernization of Traditional Chinese Medicine ; (12): 569-577, 2015.
Article in Chinese | WPRIM | ID: wpr-464811

ABSTRACT

This article was aimed to study An-shen drug substances of semen ziziphi spinosae and the relationship between index component distribution in vivo and meridian tropism. Intragastric administration of thyroid tablet suspension at the dose of 160 mg·kg-1 was given for 13 days for the establishment of yin deficiency rat model. Elevated plus maze test (EPM) was combined with light/dark box test to evaluate the effect of semen ziziphi spinosae on anxiety behavior among yin deficiency rats. The yin deficiency anxiety model rats' eyeballs were picked for blood at 10, 20, 30, 40, 60, 90, 120, 240 min after intragastric administration of semen ziziphi spinosae decoction. The rats were sacrificed for the collecting of heart, liver, spleen, lungs, kidneys, stomach, brain, large intestine and small intestine and other tissues. HPLC-PDA-ELSD was combined to detect concentrations of spinosin, jujuboside A and jujuboside B in different tissues of rats. Related pharmacokinetic parameters were achieved after processing detected data with DAS 2.0 software. The results showed that compared with the yin deficiency group, semen ziziphi spinosae significantly reduced the rats' abnormal increased food-intake (P stomach > liver > brain > large intestine > spleen > lungs > heart > kidneys. The order of AUC0-t in tissues was small intestine > stomach > liver > large intestine > spleen > brain > heart > kidneys > lungs. The order of average concentration of jujuboside A among tissues was lungs > large intestine > heart > spleen > liver > kidneys > small intestine > stomach > brain. The order of AUC0-t in tissues was lungs > spleen > liver > heart > large intestine > brain > stomach > kidneys > small intestine. The order of average concentration of jujuboside B among tissues was large intestine > small intestine > stomach. The order of AUC0-t in tissues was large intestine > small intestine > stomach. It was concluded that semen ziziphi spinosae can obviously improve yin deficiency symptoms with good anti-anxiety effects. Spinosin and jujuboside A in semen ziziphi spinosae were the drug substances of An-shen effect. They were also the material basis of sweet and sour taste. The spinosin and jujuboside A distribution in vivo of yin deficiency anxiety model rats were close to the meridian tropism of semen ziziphi spinosae.

13.
World Science and Technology-Modernization of Traditional Chinese Medicine ; (12): 1323-1328, 2013.
Article in Chinese | WPRIM | ID: wpr-441644

ABSTRACT

This study was aimed to establish an analytical method to determine spinosin, jujuboside A, jujuboside B and betulinic acid content of Ziziphi Spinosae Semen quickly and accurately. The UPLC determination of Ziziphi Spinosae Semen method was established on an HSS T3 Column (2.1 mm × 100 mm, 1.8 μm) eluted with the mobile phase consisted of water containing 0.03% phosphoric acid and acetonitrile in gradient mode with the flow rate of 0.5 mL?min-1 and the detection wavelength was set at 204 nm. The standard curves of spinosin, jujuboside A, jujuboside B, betulinic acid showed a good linearity in 29.70 - 594.0 μg?mL-1, 10.68 - 213.6 μg?mL-1, 6.175 - 123.5 μg?mL-1, 22.00 - 440.0 μg?mL-1, respectively. The mean recoveries (n = 9) of the four components were between 98 . 3% and 99 . 4%, RSD < 1 . 4%. This method which is quick and accurate can be used to determine spinosin, jujuboside A, jujuboside B and betulinic acid in Ziziphi Spinosae Semen.

14.
Chinese Journal of Information on Traditional Chinese Medicine ; (12)2006.
Article in Chinese | WPRIM | ID: wpr-581076

ABSTRACT

Objective To study the optimum process for Suanzaoren granules,and to optimize the ethanol-extracting process for jujuboside A in Semen Ziziphi Spinosae.Methods With the extracting rate of jujuboside A as the index,HPLC was used to determine the content of jujuboside A in extracted solution and the extracting conditions were optimized by orthogonal design.Results The optimum extracting condition was adding 8 times amount of 70% ethanol and refluxing for 60 min,then adding 6 times amount of 70% ethanol,refluxing and extracting for 40 min(two times).Conclusion The optimized process is stable and highly efficient.

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