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Xu C.,Shanghai University of Traditional Chinese Medicine | Chou G.-X.,Shanghai University of Traditional Chinese Medicine | Chou G.-X.,Shanghai Randnter for Standardization of Chinese Medicines | Wang C.-H.,Shanghai University of Traditional Chinese Medicine | And 2 more authors.
Phytochemistry | Year: 2012

The rare noriridoids, Andrographidoids A-E (1-5), along with a known iridoid curvifloruside F (6), were isolated from roots of Andrographis paniculata. All noriridoids were aglycones and 1-4 had (semi-) acetal structures located at C-3 but not at C-1. Their structures were established by a series of 1D and 2D NMR analyses. The antibacterial activity of these iridoids was also assessed using the microtitre plate broth dilution method. © 2011 Elsevier Ltd. All rights reserved.


Shen K.,Shanghai University of Traditional Chinese Medicine | Ji L.,Shanghai University of Traditional Chinese Medicine | Ji L.,Shanghai Randnter for Standardization of Chinese Medicines | Lu B.,Shanghai University of Traditional Chinese Medicine | And 2 more authors.
Cell Biochemistry and Biophysics | Year: 2012

c-Jun N-terminal kinase/stress-activated protein kinase (JNK/SAPK) is involved in the regulation of various cellular functions including cell cycle, proliferation, apoptosis. However, whether JNK/SAPK directly regulates the angiogenesis of human umbilical vein endothelial cells (HUVECs) induced by vascular endothelial growth factor A (VEGFA) has not yet been fully elucidated. Our present study firstly demonstrated VEGFA-induced angiogenic responses including the increase of cell viability, migration, and tube formation with a concentration-dependent manner in HUVECs. Further results showed that VEGFA induced the activation of JNK/SAPK, p38 kinase and extracellular signal-regulated kinases 1 and 2 (ERK1/2), while JNK/SAPK inhibitor SP600125 and specific siRNA both blocked all those angiogenic effects induced by VEGFA. Furthermore, VEGFA induced the phosphorylation of ASK1, SEK1/MKK4, MKK7, and c-Jun, which are upstream or downstream signals of JNK/SAPK. In addition, in vivo matrigel plug assay further showed that SP600125 inhibited VEGFA-induced angiogenesis. Further results showed that SP600125 and JNK/SAPK siRNA decreased VEGFA-induced VEGFR2 (Flk-1/KDR) sustained phosphorylation in HUVECs. Taken together, all these results demonstrate that JNK/SAPK regulates VEGFA-induced VEGFR2 sustained phosphorylation, which plays important roles in VEGFA-induced angiogenesis in HUVECs. © 2012 Springer Science+Business Media, LLC.


Chen Z.,Shanghai University of Traditional Chinese Medicine | Chen Z.,China Pharmaceutical University | Tao H.,Shanghai University of Traditional Chinese Medicine | Tao H.,Shanghai Randnter for Standardization of Chinese Medicines | And 7 more authors.
Journal of Separation Science | Year: 2014

As a final step of the purine metabolism process, xanthine oxidase catalyzes the oxidation of hypoxanthine and xanthine into uric acid. Our research has demonstrated that Erycibe obtusifolia has xanthine oxidase inhibitory properties. The purpose of this paper is to describe a new strategy based on a combination of multiple mass spectrometric platforms and thin-layer chromatography bioautography for effectively screening the xanthine oxidase inhibitory and antioxidant properties of E. obtusifolia. This strategy was accomplished through the following steps. (i) Separate the extract of E. obtusifolia into fractions by an autopurification system controlled by liquid chromatography with mass spectrometry. (ii) Determine the active fractions of E. obtusifolia by thin-layer chromatography bioautography. (iii) Identify the structure of the main active compounds with the information provided by direct analysis in real time mass spectrometry. (iv) Calculate the IC50 value of each compound against xanthine oxidase using high-performance liquid chromatography. Using the caulis of E. obtusifolia as the experimental material, seven target peaks were screened out as xanthine oxidase inhibitors or antioxidants. Our screening strategy allows for rapid analysis of small molecules with almost no sample preparation and can be completed within a week, making it a useful assay to identify unstable compounds and provide the empirical foundation for E. obtusifolia as a natural remedy for gout and oxidative-stress-related diseases. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.


Liu Q.,Shanghai Randnter for Standardization of Chinese Medicines | Chou G.-X.,Shanghai Randnter for Standardization of Chinese Medicines | Chou G.-X.,Shanghai University of Traditional Chinese Medicine | Wang Z.-T.,Shanghai Randnter for Standardization of Chinese Medicines | Wang Z.-T.,Shanghai University of Traditional Chinese Medicine
Helvetica Chimica Acta | Year: 2012

One new iridoid glucoside, 4″-O-β-D-glucosyl-6′-O-(4-O- β-D-glucosylcaffeoyl)linearoside (1), and two new secoiridoid glucosides, 6′-O-acetylsweroside (2) and 6′-O-acetyl-3′-O-[3-(β-D- glucopyranosyloxy)-2-hydroxybenzoyl]sweroside (3), were isolated from the dried roots of Gentiana manshurica (Gentianaceae), together with 11 known ones, including one iridoid glucoside, five secoiridoid glucosides, and five triterpenes. The structures of the new compounds were determined on the basis of detailed spectroscopic analyses and acidic hydrolysis. Copyright © 2012 Verlag Helvetica Chimica Acta AG, Zürich, Switzerland.


Li S.,Shanghai University of Traditional Chinese Medicine | Li S.,Xinjiang Medical University | Liu W.,Shanghai University of Traditional Chinese Medicine | Teng L.,Shanghai University of Traditional Chinese Medicine | And 7 more authors.
Journal of Pharmaceutical and Biomedical Analysis | Year: 2014

Harmane, a β-carboline alkaloid with a wide spectrum of pharmacological activities, is naturally present in the human diet, in numerous foodstuffs and in hallucinogenic plants such as Peganum harmala, Banisteriopsis caapi and Tribulus terrestris. However, the precise metabolic fate of harmane remains unknown. In order to know whether harmane is extensively metabolized, a rapid and sensitive method using ultra-performance liquid chromatography combined with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (UPLC/ESI-QTOF-MS) was used to analyze the metabolic profile of harmane in vitro and in vivo in rats. A total of 21 metabolites were identified from the rat liver microsomes and rat liver S9 (9), rat urine (11), feces (16), bile (16), and plasma (10) after a single oral administration of harmane using MetaboLynx™ and MassFragment ™ software tools. It indicated that the biliary and faecal clearance were the major excretion routes for harmane as well as its metabolites. The specific CLogP values combined with different acidic and alkaline mobile phase were helpful and useful for distinguishing N-oxidation and monohydroxylation metabolites. The metabolic transformation pathways of harmane included monohydroxylation, dihydroxylation, N-oxidation, O-glucuronide conjugation, O-sulphate conjugation, and glutathione conjugation. In conclusion, this study showed an insight into the metabolism of harmane. © 2014 Elsevier B.V.


Shen K.,Shanghai University of Traditional Chinese Medicine | Ji L.,Shanghai University of Traditional Chinese Medicine | Ji L.,Shanghai Randnter for Standardization of Chinese Medicines | Gong C.,Shanghai University of Traditional Chinese Medicine | And 7 more authors.
Biochemical Pharmacology | Year: 2012

Notoginsenoside Ft1 (Ft1) is a saponin isolated from Panax notoginseng, which has been used traditionally for the treatment of trauma injuries in East Asia. Here we show that Ft1 is a novel stimulator of angiogenesis. The results show that Ft1 induces proliferation, migration, and tube formation in cultured human umbilical vein endothelial cells (HUVECs). Ft1 increases translocalization of hypoxia-inducible factor-1α (HIF-1α) from cytoplasm to nuclei, where it binds to the vascular endothelial growth factor (VEGF) promoter, increasing the expression of VEGF mRNA and the subsequent secretion of the growth factor. Ft1 induces the activation of PI3K/AKT and Raf/MEK/ERK signaling pathways. Pharmacological inhibition with LY294002, wortmanin or PD98059 reduces Ft1-induced angiogenesis, indicating the important role played by these pathways. In addition, Ft1 induces phosphorylation of the mammalian target of rapamycin (mTOR), and siRNA-mediated mTOR knockdown decreases tube formation, proliferation, transport of HIF-1α into nuclei and VEGF mRNA expression in response to Ft1. Finally, in vivo, Ft1 promotes the formation of blood vessels in Matrigel plug and wound healing in mice. Taken together, the present results reveal that Ft1 stimulates angiogenesis via HIF-1α-mediated VEGF expression, with PI3K/AKT and Raf/MEK/ERK signaling cascades concurrently participating in the process. © 2012 Elsevier Inc. All rights reserved.


Zhao T.,Shanghai University of Traditional Chinese Medicine | Zheng S.-S.,Shanghai University of Traditional Chinese Medicine | Zhang B.-F.,Shanghai University of Traditional Chinese Medicine | Zhang B.-F.,Shanghai Randnter for Standardization of Chinese Medicines | And 6 more authors.
Food Chemistry | Year: 2012

The β-carboline alkaloids, harmaline and harmine, are present in hallucinogenic plants Ayahuasca and Peganum harmala, and in a variety of foods. In order to establish the metabolic pathway and bioactivities of endogenous and xenobiotic bioactive β-carbolines, high-performance liquid chromatography, coupled with mass spectrometry, was used to identify these metabolites in human liver microsomes (HLMs) in vitro and in rat urine and bile samples after oral administration of the alkaloids. Three metabolites of harmaline and two of harmine were found in the HLMs. Nine metabolites for harmaline and seven metabolites for harmine, from the rat urine and bile samples, were identified. Among them, four in vivo metabolites were isolated and fully characterised by NMR analysis. For the first time, harmaline is shown transforming to harmine by oxidative dehydrogenation in rat. Five metabolic pathways were therefore proposed, namely, oxidative dehydrogenation, 7-O-demethylation, hydroxylation, O-glucuronide conjugation and O-sulphate conjugation. © 2012 Elsevier Ltd. All rights reserved.


Fang L.-X.,Shanghai University of Traditional Chinese Medicine | Xiong A.-Z.,Shanghai University of Traditional Chinese Medicine | Wang R.,Shanghai University of Traditional Chinese Medicine | Wang R.,Shanghai Randnter for Standardization of Chinese Medicines | And 5 more authors.
Journal of Separation Science | Year: 2013

The objective of this study was to develop an effective strategy for screening and identifying mycotoxins in herbal medicine (HM). Here, Imperatae Rhizoma, a commonly used Chinese herb, was selected as a model HM. A crude drug contaminated with fungi was analyzed by comparing with uncontaminated ones. Ultra-performance LC coupled to tandem quadrupole TOF-MS (UPLC-Q-TOF-MS) with collision energy function was applied to analyze different samples from Imperatae Rhizoma. Then, MarkerLynxTM software was employed to screen the excess components in analytes, compared with control samples, and those selected markers were likely to be the metabolites of fungi. Furthermore, each of the accurate masses of the markers obtained from MarkerLynxTM was then searched in a mycotoxins/fungal metabolites database established in advance. The molecular formulas with relative mass error between the measured and theoretical mass within 5 ppm were chosen and then applied to MassFragmentTM analysis for further confirmation of their structures. With the use of this approach, five mycotoxins that have never been reported in HM were identified in contaminated Imperatae Rhizoma. The results demonstrate the potential of UPLC-Q-TOF-MS coupled with the MarkerLynxTM software and MassFragmentTM tool as an efficient and convenient method to screen and identify mycotoxins in herbal materials and aid in the quality control of HM. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.


Chen J.-Z.,China Pharmaceutical University | Chou G.-X.,Shanghai University of Traditional Chinese Medicine | Chou G.-X.,Shanghai Randnter for Standardization of Chinese Medicines | Wang C.-H.,Shanghai University of Traditional Chinese Medicine | And 5 more authors.
Journal of Pharmaceutical and Biomedical Analysis | Year: 2010

Norisoboldine (1,9-dihydroxy-2,10-dimethoxynoraporphine) is one of the major bioactive isoquinoline alkaloids in Linderae Radix, a commonly used Chinese herbal medicine. The aim of this study is to isolate and characterize metabolites of norisoboldine after gavage feeding in rats. High-performance liquid chromatography coupled with electrospray ionization and ion-trap mass spectrometry (HPLC-ESI/MSn) was used to identify metabolites of norisoboldine in rat urine and bile samples. A total of five metabolites of norisoboldine were characterized by comparing retention time and UV absorption in HPLC, and by molecular mass and fragmentation pattern of the analytes by mass spectrometry with those of norisoboldine. Two new glucuronide conjugates of norisoboldine, noriosboldine-1-O-β-d-glucuronide and norisoboldine-9-O-α-d-glucuronide, were isolated from rat urine samples and their structures were confirmed by NMR spectroscopy (1H, 13C, HMBC and HSQC) for the first time. The results suggested that glucuronidation and sulfation were involved in metabolic pathways of norisoboldine in rat. © 2010 Elsevier B.V.


Wang J.,Shanghai University of Traditional Chinese Medicine | Liang Q.,Shanghai University of Traditional Chinese Medicine | Ji L.,Shanghai University of Traditional Chinese Medicine | Ji L.,Shanghai Randnter for Standardization of Chinese Medicines | And 5 more authors.
Human and Experimental Toxicology | Year: 2011

The present study was undertaken to investigate the gender-related liver injury induced by Dioscorea bulbifera L. (DB), a traditional medicinal plant, in mice, and further explored its hepatotoxic chemical compound. Serum and liver tissue samples were collected at 0, 4, 8, 12 h, after mice were administrated orally with 640 mg/kg ethyl acetate extracts (EF) isolated from DB. After treatments, serum alanine transaminase (ALT) and aspartate transaminase (AST) activities were both significantly elevated. Liver lipid peroxidation (LPO) level increased, while glutathione amounts, glutathione-S-transferase (GST), superoxide dismutase (SOD) and catalase (CAT) activities all decreased in the time-dependent manner. Further analysis demonstrated that ALT and AST activities in female mice were significantly lower than those in male. Meanwhile, liver glutathione amounts and CAT activity in female mice after giving EF for 12 h were both higher than those in male. Further, comparing the liver injury induced by Diosbulbin B isolated from DB with that induced by EF on the basis of chemical analysis for the amounts of Diosbulbin B in EF of DB, we found that Diosbulbin B could be the main hepatotoxic chemical compound in DB. Taken together, our results show that DB can induce gender-related liver oxidative stress injury in mice, and its main hepatotoxic chemical compound is Diosbulbin B, for the first time. © 2011 SAGE Publications.

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