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Article ; Online: An integrated strategy by absorbed component characterization, pharmacokinetics, and activity evaluation for identification of potential nephroprotective substances in Zhu‐Ling decoction

Shu, Zhi‐heng / Fan, Cai‐lian / Wei, Hong‐yan / Li, Zi‐ting / Norimoto, Hisayoshi / Tang, Xi‐yang / Yao, Zhi‐hong / Yao, Xin‐sheng / Dai, Yi

Journal of Separation Science. 2023 Sept., v. 46, no. 17 p.e2300331-

2023

Abstract: An efficient strategy for the identification of potential nephroprotective substances in Zhu‐Ling decoction has been established with the integration of absorbed components characterization, pharmacokinetics, and activity evaluation. A qualitative method ...

Abstract	An efficient strategy for the identification of potential nephroprotective substances in Zhu‐Ling decoction has been established with the integration of absorbed components characterization, pharmacokinetics, and activity evaluation. A qualitative method was developed to characterize the chemical constituents absorbed components in vivo of Zhu‐Ling decoction by using ultra‐performance liquid chromatography‐quadrupole‐time‐of‐flight mass spectrometry. A quantitative method was established and validated for the simultaneous determination of eight compounds in rat plasma by using ultra‐performance liquid chromatography‐triple quadruple tandem mass spectrometry. Finally, the nephroprotective activities of absorbed components with high exposure were assessed by cell survival rate, superoxide dismutase, and malondialdehyde activities in hydrogen peroxide–induced Vero cells. As a result, 111 compounds in Zhu‐Ling decoction and 36 absorbed components were identified in rat plasma and urine, and poricoic acid A, poricoic acid B, alisol A, 16‐oxo‐alisol A, and dehydro‐tumulosic acid had high exposure levels in rat plasma. Finally, poricoic acid B, poricoic acid A, 16‐oxo‐alisol A, and dehydro‐tumulosic acid showed remarkable nephroprotective activity against Vero cells damage induced by hydrogen peroxide. Besides, superoxide dismutase and malondialdehyde activities were obviously regulated in hydrogen peroxide–induced Vero cells by treatment with the four compounds mentioned above. Therefore, these four compounds were considered to be effective substances of Zhu‐Ling decoction due to their relatively high exposure in vivo and biological activity. This study provided a chemical basis for the action mechanism of Zhu‐Ling decoction in the treatment of chronic kidney diseases.
Keywords	cell viability ; hydrogen peroxide ; kidneys ; malondialdehyde ; nephroprotective effect ; pharmacokinetics ; quantitative analysis ; rats ; separation ; superoxide dismutase ; survival rate ; tandem mass spectrometry ; urine
Language	English
Dates of publication	2023-09
Publishing place	John Wiley & Sons, Ltd
Document type	Article ; Online
Note	JOURNAL ARTICLE
ZDB-ID	2047990-6
ISSN	1615-9314 ; 1615-9306
ISSN (online)	1615-9314
ISSN	1615-9306
DOI	10.1002/jssc.202300331
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Chemical and metabolic profiling of Codonopsis Radix extract with an integrated strategy using ultra‐high‐performance liquid chromatography coupled with mass spectrometry

Tang, Xi‐Yang / Zeng, Jia‐Xing / Wang, Xiao‐Xing / Xu, Wan‐Yi / Zhao, Peng‐Cheng / Fan, Cai‐Lian / Yao, Zhi‐Hong / Yao, Xin‐Sheng / Dai, Yi

Journal of Separation Science. 2023 Jan., v. 46, no. 2 p.e2200723-

2023

Abstract: Codonopsis radix was commonly used as food materials or herbal medicines in many countries. However, the comprehensive analysis of chemical constituents, and in vivo xenobiotics of Codonopsis radix remain unclear. In the present study, an integrated ... ...

Abstract	Codonopsis radix was commonly used as food materials or herbal medicines in many countries. However, the comprehensive analysis of chemical constituents, and in vivo xenobiotics of Codonopsis radix remain unclear. In the present study, an integrated strategy with feature‐based molecular networking using ultra‐high‐performance liquid chromatography coupled with mass spectrometry was established to systematically screen the chemical constituents and the in vivo xenobiotics of Codonopsis radix. A step‐by‐step manner based on a composition database, visual structure classification, discriminant ions, and metabolite software prediction was proposed to overcome the complexities due to the similar structure of chemical constituents and metabolites of Codonopsis radix. As a result, 103 compounds were tentatively characterized, 20 of which were identified by reference standards. Besides, a total of 50 xenobiotics were detected in vivo, including 26 prototypes and 24 metabolites, while the metabolic features of the pyrrolidine alkaloids were elucidated for the first time. The metabolism reactions of pyrrolidine alkaloids and sesquiterpene lactones included oxidation, methylation, hydration, hydrogenation, demethylation, glucuronidation, and sulfation. This study provided a generally applicable approach to the comprehensive investigation of the chemical and metabolic profile of traditional Chinese medicine and offered reasonable guidelines for further screening of quality control indicators and pharmacodynamics mechanism of Codonopsis radix.
Keywords	Codonopsis ; Oriental traditional medicine ; computer software ; databases ; demethylation ; hydrogenation ; mass spectrometry ; metabolites ; methylation ; oxidation ; pharmacodynamics ; prediction ; quality control ; separation ; sesquiterpenoid lactones ; ultra-performance liquid chromatography ; xenobiotics
Language	English
Dates of publication	2023-01
Publishing place	John Wiley & Sons, Ltd
Document type	Article ; Online
Note	JOURNAL ARTICLE
ZDB-ID	2047990-6
ISSN	1615-9314 ; 1615-9306
ISSN (online)	1615-9314
ISSN	1615-9306
DOI	10.1002/jssc.202200723
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Pharmacokinetics, hepatic disposition, and heart tissue distribution of 14 compounds in rat after oral administration of Qi-Li-Qiang-Xin capsule via ultra-high-performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry.

Tang, Xi-Yang / Dai, Zi-Qin / Zeng, Jia-Xing / Li, Zi-Ting / Fan, Cai-Lian / Yao, Zhi-Hong / Yao, Xin-Sheng / Dai, Yi

Journal of separation science

2022 Volume 45, Issue 13, Page(s) 2177–2189

Abstract: In the present study, a specific and sensitive approach using ultra-high-performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry was developed and validated for the quantitative analysis of 14 constituents in rat plasma, ...

Abstract	In the present study, a specific and sensitive approach using ultra-high-performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry was developed and validated for the quantitative analysis of 14 constituents in rat plasma, liver, and heart. The method was fully validated and successfully applied to pharmacokinetic, hepatic disposition, and heart tissue distribution studies of 14 compounds after the oral administration of Qi-Li-Qiang-Xin capsule. Ginsenoside Rb1, alisol A, astragaloside IV, and periplocymarin were found to be highly exposed in rat plasma, while toxic components such as hypaconitine, mesaconitine, and periplocin had low circulation levels in vivo. Moreover, sinapine thiocyanate, neoline, formononetin, calycosin, and alisol A exhibited significant liver first-pass effects. Notably, high levels of alisol A, periplocymarin, benzoylmesaconine, and benzoylhypaconine were observed in the heart. Based on high exposure and appropriate pharmacokinetic features in the systemic plasma and heart, astragaloside IV, ginsenoside Rb1, periplocymarin, benzoylmesaconine, benzoylhypaconine, and alisol A can be considered as the main potentially effective components. Ultimately, the results provide relevant information for discovery of effective substances, as well as further anti-heart failure action mechanism investigations of Qi-Li-Qiang-Xin capsule.
MeSH term(s)	Administration, Oral ; Animals ; Chromatography, High Pressure Liquid/methods ; Drugs, Chinese Herbal/analysis ; Liver/chemistry ; Rats ; Tandem Mass Spectrometry/methods ; Tissue Distribution
Chemical Substances	Drugs, Chinese Herbal
Language	English
Publishing date	2022-05-03
Publishing country	Germany
Document type	Journal Article
ZDB-ID	2047990-6
ISSN	1615-9314 ; 1615-9306
ISSN (online)	1615-9314
ISSN	1615-9306
DOI	10.1002/jssc.202101008
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Pharmacokinetics, hepatic disposition, and heart tissue distribution of 14 compounds in rat after oral administration of Qi‐Li‐Qiang‐Xin capsule via ultra‐high‐performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry

Tang, Xi‐Yang / Dai, Zi‐Qin / Zeng, Jia‐Xing / Li, Zi‐Ting / Fan, Cai‐Lian / Yao, Zhi‐Hong / Yao, Xin‐Sheng / Dai, Yi

Journal of separation science. 2022 July, v. 45, no. 13

2022

Abstract: In the present study, a specific and sensitive approach using ultra‐high‐performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry was developed and validated for the quantitative analysis of 14 constituents in rat plasma, ...

Abstract	In the present study, a specific and sensitive approach using ultra‐high‐performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry was developed and validated for the quantitative analysis of 14 constituents in rat plasma, liver, and heart. The method was fully validated and successfully applied to pharmacokinetic, hepatic disposition, and heart tissue distribution studies of 14 compounds after the oral administration of Qi‐Li‐Qiang‐Xin capsule. Ginsenoside Rb1, alisol A, astragaloside IV, and periplocymarin were found to be highly exposed in rat plasma, while toxic components such as hypaconitine, mesaconitine, and periplocin had low circulation levels in vivo. Moreover, sinapine thiocyanate, neoline, formononetin, calycosin, and alisol A exhibited significant liver first‐pass effects. Notably, high levels of alisol A, periplocymarin, benzoylmesaconine, and benzoylhypaconine were observed in the heart. Based on high exposure and appropriate pharmacokinetic features in the systemic plasma and heart, astragaloside IV, ginsenoside Rb1, periplocymarin, benzoylmesaconine, benzoylhypaconine, and alisol A can be considered as the main potentially effective components. Ultimately, the results provide relevant information for discovery of effective substances, as well as further anti‐heart failure action mechanism investigations of Qi‐Li‐Qiang‐Xin capsule.
Keywords	astragalosides ; formononetin ; ginsenosides ; heart ; liver ; oral administration ; pharmacokinetics ; quantitative analysis ; rats ; separation ; sinapine ; tandem mass spectrometry ; thiocyanates ; tissue distribution ; toxicity ; ultra-performance liquid chromatography
Language	English
Dates of publication	2022-07
Size	p. 2177-2189.
Publishing place	John Wiley & Sons, Ltd
Document type	Article
Note	JOURNAL ARTICLE
ZDB-ID	2047990-6
ISSN	1615-9314 ; 1615-9306
ISSN (online)	1615-9314
ISSN	1615-9306
DOI	10.1002/jssc.202101008
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Article ; Online: An integrated strategy by absorbed component characterization, pharmacokinetics, and activity evaluation for identification of potential nephroprotective substances in Zhu-Ling decoction.

Shu, Zhi-Heng / Fan, Cai-Lian / Wei, Hong-Yan / Li, Zi-Ting / Norimoto, Hisayoshi / Tang, Xi-Yang / Yao, Zhi-Hong / Yao, Xin-Sheng / Dai, Yi

Journal of separation science

2023 Volume 46, Issue 17, Page(s) e2300331

Abstract: An efficient strategy for the identification of potential nephroprotective substances in Zhu-Ling decoction has been established with the integration of absorbed components characterization, pharmacokinetics, and activity evaluation. A qualitative method ...

Abstract	An efficient strategy for the identification of potential nephroprotective substances in Zhu-Ling decoction has been established with the integration of absorbed components characterization, pharmacokinetics, and activity evaluation. A qualitative method was developed to characterize the chemical constituents absorbed components in vivo of Zhu-Ling decoction by using ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry. A quantitative method was established and validated for the simultaneous determination of eight compounds in rat plasma by using ultra-performance liquid chromatography-triple quadruple tandem mass spectrometry. Finally, the nephroprotective activities of absorbed components with high exposure were assessed by cell survival rate, superoxide dismutase, and malondialdehyde activities in hydrogen peroxide-induced Vero cells. As a result, 111 compounds in Zhu-Ling decoction and 36 absorbed components were identified in rat plasma and urine, and poricoic acid A, poricoic acid B, alisol A, 16-oxo-alisol A, and dehydro-tumulosic acid had high exposure levels in rat plasma. Finally, poricoic acid B, poricoic acid A, 16-oxo-alisol A, and dehydro-tumulosic acid showed remarkable nephroprotective activity against Vero cells damage induced by hydrogen peroxide. Besides, superoxide dismutase and malondialdehyde activities were obviously regulated in hydrogen peroxide-induced Vero cells by treatment with the four compounds mentioned above. Therefore, these four compounds were considered to be effective substances of Zhu-Ling decoction due to their relatively high exposure in vivo and biological activity. This study provided a chemical basis for the action mechanism of Zhu-Ling decoction in the treatment of chronic kidney diseases.
MeSH term(s)	Chlorocebus aethiops ; Rats ; Animals ; Hydrogen Peroxide ; Vero Cells ; Mass Spectrometry/methods ; Triterpenes ; Drugs, Chinese Herbal/pharmacology ; Drugs, Chinese Herbal/chemistry ; Chromatography, High Pressure Liquid/methods
Chemical Substances	poricoic acid A ; Hydrogen Peroxide (BBX060AN9V) ; Triterpenes ; Drugs, Chinese Herbal
Language	English
Publishing date	2023-07-12
Publishing country	Germany
Document type	Journal Article
ZDB-ID	2047990-6
ISSN	1615-9314 ; 1615-9306
ISSN (online)	1615-9314
ISSN	1615-9306
DOI	10.1002/jssc.202300331
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Clinical Research on the Mechanisms Underlying Immune Checkpoints and Tumor Metastasis.

Tang, Xi-Yang / Shi, An-Ping / Xiong, Yan-Lu / Zheng, Kai-Fu / Liu, Yu-Jian / Shi, Xian-Gui / Jiang, Tao / Zhao, Jin-Bo

Frontiers in oncology

2021 Volume 11, Page(s) 693321

Abstract: This study highlights aspects of the latest clinical research conducted on the relationship between immune checkpoints and tumor metastasis. The overview of each immune checkpoint is divided into the following three sections: 1) structure and expression; ...

Abstract	This study highlights aspects of the latest clinical research conducted on the relationship between immune checkpoints and tumor metastasis. The overview of each immune checkpoint is divided into the following three sections: 1) structure and expression; 2) immune mechanism related to tumor metastasis; and 3) clinical research related to tumor metastasis. This review expands on the immunological mechanisms of 17 immune checkpoints, including TIM-3, CD47, and OX-40L, that mediate tumor metastasis; evidence shows that most of these immune checkpoints are expressed on the surface of T cells, which mainly exert immunomodulatory effects. Additionally, we have summarized the roles of these immune checkpoints in the diagnosis and treatment of metastatic tumors, as these checkpoints are considered common predictors of metastasis in various cancers such as prostate cancer, non-Hodgkin lymphoma, and melanoma. Moreover, certain immune checkpoints can be used in synergy with PD-1 and CTLA-4, along with the implementation of combination therapies such as LIGHT-VTR and anti-PD-1 antibodies. Presently, most monoclonal antibodies generated against immune checkpoints are under investigation as part of ongoing preclinical or clinical trials conducted to evaluate their efficacy and safety to establish a better combination treatment strategy; however, no significant progress has been made regarding monoclonal antibody targeting of CD28, VISTA, or VTCN1. The application of immune checkpoint inhibitors in early stage tumors to prevent tumor metastasis warrants further evidence; the immune-related adverse events should be considered before combination therapy. This review aims to elucidate the mechanisms of immune checkpoint and the clinical progress on their use in metastatic tumors reported over the last 5 years, which may provide insights into the development of novel therapeutic strategies that will assist with the utilization of various immune checkpoint inhibitors.
Language	English
Publishing date	2021-07-22
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2649216-7
ISSN	2234-943X
ISSN	2234-943X
DOI	10.3389/fonc.2021.693321
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: A new strategy for discovering effective substances and mechanisms of traditional Chinese medicine based on standardized drug containing plasma and the absorbed ingredients composition, a case study of Xian-Ling-Gu-Bao capsules.

Qiu, Zuo-Cheng / Tang, Xi-Yang / Wu, Qing-Chang / Tang, Zi-Ling / Wong, Man-Sau / Chen, Jia-Xu / Yao, Xin-Sheng / Dai, Yi

Journal of ethnopharmacology

2021 Volume 279, Page(s) 114396

Abstract: Ethnopharmacological relevance: The overall therapeutic effect of traditional Chinese medicine formulae (TCMF) was achieved by the interactions of multiple components with multiple targets. However, current pharmacology research strategies have ... ...

Abstract	Ethnopharmacological relevance: The overall therapeutic effect of traditional Chinese medicine formulae (TCMF) was achieved by the interactions of multiple components with multiple targets. However, current pharmacology research strategies have struggled to identify effective substance groups and encountered challenges in elucidating the underlying mechanisms of TCMF. Aim: In this study, a comprehensive strategy was proposed and applied to elucidate the interactions of the multiple components that underlie the functions of the famous TCMF: Xian-Ling-Gu-Bao (XLGB) capsule on bone metabolism in vivo and to elucidate the molecular mechanisms underlying the effects of XLGB on bone cells, especially on osteoblasts. Methods: The efficacy of XLGB in the protection against bones loss in ovariectomized (OVX) rats was confirmed by Micro-CT analysis. The anti-osteoporosis mechanism involved in the systemic regulatory actions of XLGB was elucidated by transcriptome sequencing analysis on bone marrow mesenchymal stem cells isolated from OVX rats. Moreover, the components absorbed in XLGB-treated plasma were characterized by mass spectrometry analysis, and subsequently, a standardized preparation process of drug-containing plasma was established. The synergistic osteogenic effect of the multiple components in plasma was investigated by a combination and then knockout of components using pre-osteoblast MC3T3-E1 cells. In order to decipher the underlying mechanism of XLGB, the targets of the absorbed components on bone were predicted by target prediction and network pharmacology analysis, then several interactions were validated by biochemical and cell-based assay. Results: A total of 18 genes, including HDC, CXCL1/2, TNF, IL6 and Il1b, were newly found to be the major target genes regulated by XLGB. Interestingly, we found that a combination of the three absorbed components, i.e. MSP, rather than their single form at the same concentration, stimulated the formation of calcified nodules in MC3T3-E1 cells, suggesting a synergistic effect of these components. Besides, target prediction and experimental validation confirmed the binding affinity of corylin and icaritin for estrogen receptor α and β, the inhibitory activity of isobavachin and isobavachalcone on glycogen synthase kinase-3β, and the inhibitory activity of isobavachalcone on cathepsin K. The cell-based assay further confirmed the result of the biochemical assay. A network that integrated absorbed components of XLGB-targets-perturbation genes-pathways against osteoporosis was established. Conclusion: Our current study provides a new systemic strategy for discovering active ingredient groups of TCM formulae and understanding their underlying mechanisms.
MeSH term(s)	3T3 Cells ; Administration, Oral ; Animals ; Bone Density/drug effects ; Bone Marrow Cells ; Cell Differentiation/drug effects ; Cell Proliferation/drug effects ; Drugs, Chinese Herbal/therapeutic use ; Estradiol/pharmacology ; Female ; Gene Expression Regulation/drug effects ; Gene Regulatory Networks ; Medicine, Chinese Traditional ; Mice ; Osteoblasts/drug effects ; Osteoblasts/physiology ; Osteoporosis/prevention & control ; Ovariectomy ; RANK Ligand/pharmacology ; RAW 264.7 Cells ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Signal Transduction/drug effects ; Stem Cells
Chemical Substances	Drugs, Chinese Herbal ; RANK Ligand ; Tnfsf11 protein, mouse ; xian ling gu bao ; Estradiol (4TI98Z838E)
Language	English
Publishing date	2021-07-08
Publishing country	Ireland
Document type	Journal Article
ZDB-ID	134511-4
ISSN	1872-7573 ; 0378-8741
ISSN (online)	1872-7573
ISSN	0378-8741
DOI	10.1016/j.jep.2021.114396
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Zs.A 1494: Show issues

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Article ; Online: Effects of Xian-Ling-Gu-Bao capsule on the gut microbiota in ovariectomized rats: Metabolism and modulation.

Tang, Xi-Yang / Gao, Meng-Xue / Xiao, Hui-Hui / Dai, Zi-Qin / Yao, Zhi-Hong / Dai, Yi / Yao, Xin-Sheng

Journal of chromatography. B, Analytical technologies in the biomedical and life sciences

2021 Volume 1176, Page(s) 122771

Abstract: Xian-Ling-Gu-Bao capsule (XLGB) has been proven to prevent and treat osteoporosis. However, as a long-term oral formula, XLGB's effects on the metabolic capacity, structure and function of gut microbiota have yet to be elucidated in ovariectomized (OVX) ... ...

Abstract	Xian-Ling-Gu-Bao capsule (XLGB) has been proven to prevent and treat osteoporosis. However, as a long-term oral formula, XLGB's effects on the metabolic capacity, structure and function of gut microbiota have yet to be elucidated in ovariectomized (OVX) rats. Our objectives were to evaluate the capacity of gut microbiota for metabolizing XLGB ingredients and to assess the effect of this prescription on gut microbiota. Herein, an integrated analysis that combined ultrahigh-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) and ultrahigh-performance liquid chromatography tandem triple quadrupole mass spectrometry (UPLC-TQD-MS) was conducted to determine the metabolic capacity of gut microbiota. The effects of XLGB on gut microbiota were explored by metagenomic sequencing in OVX rats. Fecal samples from each group were collected after intragastric administration for three months. In total, 64 biotransformation products were fully characterized with rat gut microbiota from the OVX group and the XLGB group. The deglycosylation reaction was the main biotransformation pathway in core structures in the group that was incubated with XLGB. Compared with the OVX group, different biotransformation products and pathways of the XLGB group after incubation for 2 h and 8 h were described. After three months of feeding with XLGB, the domesticated gut microbiota was conducive to the production of active absorbed components via deglycosylation, such as icaritin, psoralen and isopsoralen. Comparisons of the gut microbiota of the OVX and XLGB groups showed differences in the relative abundances of the two dominant bacterial divisions, namely, Firmicutes and Bacteroidetes. The proportion of Firmicutes was significantly lower and that of Bacteroidetes was significantly higher in the XLGB group. This result demonstrated that XLGB could provide a basis for the treatment of osteoporosis by regulating lipid and bile acid metabolism. In addition, the increase in Lactobacillus, Bacteroides and Prevotella could be an important factor that led to easier production of active absorbed aglycones in the XLGB group. Our observation provided further evidence of the importance of gut microbiota in the metabolism and potential activity of XLGB.
Language	English
Publishing date	2021-05-18
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	1180823-8
ISSN	1873-376X ; 0378-4347 ; 1570-0232 ; 1387-2273
ISSN (online)	1873-376X
ISSN	0378-4347 ; 1570-0232 ; 1387-2273
DOI	10.1016/j.jchromb.2021.122771
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Zs.A 813: Show issues

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Article ; Online: An UHPLC-MS/MS method for simultaneous determination of ten sex steroid hormones in ovariectomy-induced osteoporosis rat and its application in discovery of sex steroid hormones regulatory components of Xian-Ling-Gu-Bao capsule.

Tang, Xi-Yang / Dai, Zi-Qin / Shi, Dan-Feng / Zeng, Jia-Xing / Wang, Xin-Luan / Li, Ling / Yao, Xin-Sheng / Dai, Yi

Journal of pharmaceutical and biomedical analysis

2021 Volume 195, Page(s) 113888

Abstract: Sex steroid hormones could directly affect the bone metabolism by regulating cell physiological functions. In female, it inevitably causes the abnormal levels of sex steroid hormones at post-menopause in vivo. Ovariectomized rats and mice are classic ... ...

Abstract	Sex steroid hormones could directly affect the bone metabolism by regulating cell physiological functions. In female, it inevitably causes the abnormal levels of sex steroid hormones at post-menopause in vivo. Ovariectomized rats and mice are classic animal models of osteoporosis to better understand the action mechanism of anti-osteoporosis drugs. However, it is not clear whether Xian-Ling-Gu-Bao capsule (XLGB), a kidney-tonifying traditional Chinese medicine prescription, treat osteoporosis via regulating multiple sex steroid hormones. In the present study, a reliable method involving ultra high-performance liquid chromatography coupled with triple quadrupole mass spectrometry (UHPLC/TQ-XS-MS) was developed for simultaneous quantitative analysis of ten sex steroid hormones (three estrogens, five androgens and two progestogens) in rat and mouse serum. The results of methodology were acceptable. The validated method was then successfully applied in the determination of the levels of sex steroid hormones in ovariectomy-induced osteoporosis rats, as well as drug (17β-estradiol and XLGB) intervened rats. As a result, XLGB could not only significantly increase the level of 17β-estradiol, but also improve the levels of progesterone, 17α-hydroxyprogesterone and androstenedione. Combined with molecular docking results and pharmacokinetic parameters, psoralen, isopsoralen and sweroside were considered as the key effective components of XLGB to activate adenylyl cyclase on promoting the biosynthesis of multiple sex steroid hormones. It is the first time to evaluate the regulatory effect of kidney-tonifying traditional Chinese medicine prescription on the levels of steroids in ovariectomy-induced osteoporosis rat, as well as the potential substance basis and mechanism of steroid hormone regulation.
MeSH term(s)	Animals ; Chromatography, High Pressure Liquid ; Female ; Gonadal Steroid Hormones ; Humans ; Mice ; Molecular Docking Simulation ; Osteoporosis/drug therapy ; Osteoporosis/etiology ; Ovariectomy ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry
Chemical Substances	Gonadal Steroid Hormones
Language	English
Publishing date	2021-01-02
Publishing country	England
Document type	Journal Article
ZDB-ID	604917-5
ISSN	1873-264X ; 0731-7085
ISSN (online)	1873-264X
ISSN	0731-7085
DOI	10.1016/j.jpba.2020.113888
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Zs.A 1850: Show issues

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Article: Sweroside promotes osteoblastic differentiation and mineralization via interaction of membrane estrogen receptor-α and GPR30 mediated p38 signalling pathway on MC3T3-E1 cells

Wu, Qing-chang / Tang, Xi-yang / Dai, Zi-qin / Dai, Yi / Xiao, Hui-hui / Yao, Xin-sheng

Phytomedicine. 2020 Mar., v. 68

2020

Abstract: Dipsaci Radix has been clinically used for thousands of years in China for strengthening muscles and bones. Sweroside is the major active iridoid glycoside isolated from Dipsaci Radix. It has been reported that sweroside can promote alkaline phosphatase ( ...

Abstract	Dipsaci Radix has been clinically used for thousands of years in China for strengthening muscles and bones. Sweroside is the major active iridoid glycoside isolated from Dipsaci Radix. It has been reported that sweroside can promote alkaline phosphatase (ALP) activity in both the human osteosarcoma cell line MG-63 and rat osteoblasts. However, the underlying mechanism involved in these osteoblastic processes is poorly understood.This study aimed to characterize the bone protective effects of sweroside and to investigate the signaling pathway that is involved in its actions in MC3T3-E1 cells.Cell proliferation, differentiation and mineralization were evaluated by the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay, ALP test and Alizarin Red S staining, respectively. The concentration of sweroside in intracellular and extracellular fluids was determined by ultra-performance liquid chromatography coupled to triple quadrupole xevo-mass spectrometry (UPLC/TQ-XS-MS). Proteins associated with the osteoblastic signaling pathway were analysed by western blot and immunofluorescence methods.Sweroside did not obviously affect the proliferation but significantly promoted the ALP activity and mineralization of MC3T3-E1 cells. The maximal absorption amount 0.465 ng/ml (1.3 × 10−9 M) of sweroside was extremely lower than the tested concentration of 358.340 ng/ml (10−6 M), indicating an extremely low absorption rate by MC3T3-E1 cells. Moreover, the ALP activity, the protein expression of ER-α and G protein-coupled receptor 30 (GPR30) induced by sweroside were markedly blocked by both the ER antagonist ICI 182780 and the GPR30 antagonist G15. In addition, sweroside also activated the phosphorylation of p38 kinase (p-p38), while the phosphorylation effects together with ALP and mineralization activities were completely blocked by a p38 antagonist, SB203580. Additionally, the phosphorylation of p38 induced by sweroside were markedly blocked by both the ER antagonist ICI 182780 and the GPR30 antagonist G15.The present study indicated that sweroside, as a potential agent in treatment of osteoporosis, might exert beneficial effects on MC3T3-E1 cells by interaction with the membrane estrogen receptor-α and GPR30 that then activates the p38 signaling pathway. This is the first study to report the specific mechanism of the effects of sweroside on osteoblastic differentiation and mineralization of MC3T3-E1 cells.
Keywords	G-protein coupled receptors ; Western blotting ; alizarin ; alkaline phosphatase ; antagonists ; bones ; estrogens ; extracellular fluids ; fluorescent antibody technique ; human diseases ; iridoid glycosides ; mineralization ; muscles ; osteoblasts ; osteoporosis ; osteosarcoma ; phosphorylation ; protective effect ; protein synthesis ; signal transduction ; spectroscopy ; staining ; China
Language	English
Dates of publication	2020-03
Publishing place	Elsevier GmbH
Document type	Article
ZDB-ID	1205240-1
ISSN	1618-095X ; 0944-7113
ISSN (online)	1618-095X
ISSN	0944-7113
DOI	10.1016/j.phymed.2019.153146
Database	NAL-Catalogue (AGRICOLA)

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Zs.A 4115: Show issues

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