1.Molecular cloning and functional characterization of an isoflavone glucosyltransferase from Pueraria thomsonii.
Hai-Yan DUAN ; Jian WANG ; Liang-Ping ZHA ; Hua-Sheng PENG ; Yu-Ping ZHAO ; Yuan YUAN ; Lu-Qi HUANG
Chinese Journal of Natural Medicines (English Ed.) 2022;20(2):133-138
Pueraria thomsonii has long been used in traditional Chinese medicine. Isoflavonoids are the principle pharmacologically active components, which are primarily observed as glycosyl-conjugates and accumulate in P. thomsonii roots. However, the molecular mechanisms underlying the glycosylation processes in (iso)flavonoid biosynthesis have not been thoroughly elucidated. In the current study, an O-glucosyltransferase (PtUGT8) was identified in the medicinal plant P. thomsonii from RNA-seq database. Biochemical assays of the recombinant PtUGT8 showed that it was able to glycosylate chalcone (isoliquiritigenin) at the 4-OH position and glycosylate isoflavones (daidzein, formononetin, and genistein) at the 7-OH or 4'-OH position, exhibiting no enzyme activity to flavonones (liquiritigenin and narigenin) in vitro. The identification of PtUGT8 may provide a useful enzyme catalyst for efficient biotransformation of isoflavones and other natural products for food or pharmacological applications.
Cloning, Molecular
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Genistein
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Glucosyltransferases/metabolism*
;
Isoflavones/pharmacology*
;
Pueraria/chemistry*
2.Role of PPAR-γ-regulated autophagy in genistein-induced inhibition of hepatic stellate cell activation.
Xipeng LIU ; Meifang ZHANG ; Haifeng ZHANG ; Anda ZHAO ; Juan SUN ; Wen TANG
Journal of Southern Medical University 2019;39(5):561-565
OBJECTIVE:
To investigate the inhibitory effect of genistein on activation of hepatic stellate cells (HSCs) and the role of the autophagy pathway regulated by PPAR-γ in mediating this effect.
METHODS:
Cultured HSC-T6 cells were exposed to different concentrations of genistein for 48 h, and HSC activation was verified by detecting the expressions of -SMA and 1(I) collagen; autophagy activation in the cells was determined by detecting the expressions of LC3-II and p62 using Western blotting. The autophagy inhibitor 3-MA was used to confirm the role of autophagy in genistein-induced inhibition of HSC activation. A PPAR-γ inhibitor was used to explore the role of PPAR-γ in activating autophagy in the HSCs.
RESULTS:
Genistein at concentrations of 5 and 50 μmol/L significantly inhibited the expressions of -SMA and 1(I) collagen ( < 0.05), markedly upregulated the expressions of PPAR-γ and the autophagy-related protein LC3-II ( < 0.05) and significantly down-regulated the expression of the ubiqutin-binding protein p62 ( < 0.05) in HSC-T6 cells. The cells pretreated with 3-MA prior to genistein treatment showed significantly increased protein expressions of -SMA and 1(I) collagen compared with the cells treated with genistein only ( < 0.05). Treatment with the PPAR-γ inhibitor obviously lowered the expression of LC3-II and enhanced the expression p62 in genistein-treated HSC-T6 cells, suggesting the activation of the autophagy pathway.
CONCLUSIONS
PPAR-γ- regulated autophagy plays an important role in mediating genistein-induced inhibition of HSC activation .
Anticarcinogenic Agents
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pharmacology
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Autophagy
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Collagen Type I
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Genistein
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pharmacology
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Hepatic Stellate Cells
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Humans
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PPAR gamma
;
physiology
3.A new isoflavone from Dalbergia odorifera and inhibitory activity of its tyrosinase.
Suet CHEUNG ; Wan FANG ; Xiao-Qing LI ; Rui WANG ; Shi-Kai YAN ; Hui-Zi JIN
China Journal of Chinese Materia Medica 2022;47(18):4959-4965
Twelve flavonoids were isolated and purified from the ethyl acetate fraction of 95% ethanol extract of Dalbergia odorifera by heat reflux extraction, solvent extraction, recrystallization, normal phase silica gel, Sephadex LH-20, MCI gel and HPLC methods. The structures were identified with multiple spectroscopic methods, including 1 D-NMR, 2 D-NMR and MS. The compounds were identified as 6,7,8-trimethoxy-5,4'-dihydroxy isoflavone(1), medicarpin(2), 7,2'-dihydroxy-4'-methoxy-isoflavanol(3), biochanin A(4), prunetin(5), genistein(6), pratensein(7), 3-(4-hydroxyphenyl)-6-isopentenyl-7-methoxy-4H-chromen-4-one(8), tectorigenin(9), irisolidone(10), vestitol(11), and formononetin(12). Compound 1 was a new isoflavone, and compound 8 was isolated from D. odorifera for the first time. The results showed that compounds 1-3 had inhibitory effects on tyrosinase, with inhibition rates of 35.58%, 38.63% and 51.34% at the concentration of 1.0 mmol·L~(-1), respectively.
Dalbergia/chemistry*
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Ethanol
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Flavonoids/chemistry*
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Genistein
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Isoflavones/pharmacology*
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Monophenol Monooxygenase
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Plant Extracts/pharmacology*
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Silica Gel
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Solvents
4.Molecular docking of epidermal growth factor receptor tyramine kinase domain and its inhibitor genistein.
Jianglan YUAN ; Hui LIU ; Xu KANG ; Guolin ZOU
Chinese Journal of Biotechnology 2008;24(10):1813-1817
Genistein is a high specific and noncompetitive inhibitor of epidermal growth factor receptor tyramine kinase domain (EGFR-TK). In the paper, a molecular docking between genistein and EGFR-TK was studied to explore the mechanism of their interaction and antitumor mechanism of genistein by AUTODOCK 3.05 program. The results indicated that genistein located in the active cavity of EGFR-TK by high affinity (deltaG = -31.2 kJ/mol), and genistein inhibited EGFR-TK by interfering with forming of Lys721/Glu738 ion pair. The inhibition belonged to noncompetitive interaction, in which hydrophobic force and hydrogen bond played key roles.
Catalytic Domain
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Genistein
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metabolism
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pharmacology
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Models, Molecular
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Receptor, Epidermal Growth Factor
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antagonists & inhibitors
;
metabolism
6.Chemical constituents from Artocarpus incisus and their inhibitory effects on proliferation of synoviocytes in vitro.
Yu-Tong XIE ; Shu-Hong XIONG ; Yuan BIAN ; Yu WANG ; Ruo-Qing GUAN ; Xin-Yuan SUO ; Meng-Ran DU ; Yan-Ping LIU ; Yan-Hui FU
China Journal of Chinese Materia Medica 2022;47(17):4665-4673
The chemical constituents from the branches and leaves of Artocarpus incisus were isolated and purified via silica gel, ODS, and Sephadex LH-20 column chromatography as well as preparative HPLC. The chemical structures of all isolated compounds were identified in the light of their physicochemical properties, spectroscopic analyses, and comparisons of their physicochemical and spectroscopic data with the reported data in literature. As a result, 20 compounds were isolated and characterized from the 90% ethanol extract of the branches and leaves of A. incisus, which were identified as tephrosin(1), 6-hydroxy-6 a, 12 a-dehydrodeguelin(2), sarcolobin(3), lupiwighteone(4), 12-deoxo-12α-methoxyelliptone(5), 6 aα,12 aα-12 a-hydroxyelliptone(6), homopterocarpin(7), 3-hydroxy-8,9-dimethoxypterocarpan(8), pterocarpin(9), maackiain(10), medicarpin(11), calycosin(12), genistein(13), formononetin(14), 5-hydroxy-4',7-dimethoxy isoflavone(15), liquiritigenin(16), 4(15)-eudesmene-1β,7α-diol(17), ent-4(15)-eudesmene-1β,6α-diol(18), 1α-hydroxyisodauc-4-en-15-al(19), and guaianediol(20). Except compounds 13 and 16, all other compounds were isolated from the Artocarpus plants for the first time. Additionally, using MTS assay, compounds 1-20 were eva-luated for their anti-rheumatoid arthritis activities by measuring their anti-proliferative effects on synoviocytes in vitro. As a consequence, compounds 1-16 showed notable anti-rheumatoid arthritis activities, which displayed inhibitory effects on the proliferation of MH7 A synovial fibroblast cells, with the IC_(50) values in range of(9.86±0.09)-(218.07±1.96) μmol·L~(-1).
Arthritis
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Artocarpus
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Cell Proliferation
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Ethanol
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Genistein
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Plant Extracts/pharmacology*
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Silica Gel
;
Synoviocytes
7.Effects of tyrosine protein kinase inhibitor genistein on the cell cycles of activated hepatic stellate cells.
Qiong WANG ; Yi-ping WANG ; Xiao-jing LIU ; Ming-hui HUANG
Chinese Journal of Hepatology 2003;11(1):53-53
Animals
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Cell Cycle
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drug effects
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Enzyme Inhibitors
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pharmacology
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Genistein
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pharmacology
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Liver
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cytology
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drug effects
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Male
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Rats
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Rats, Wistar
9.The covalent binding of genistein to the non-prosthetic-heme-moiety of bovine lactoperoxidase leads to enzymatic inactivation.
Hebron C CHANG ; Daniel R DOERGE ; Chenghong HSIEH ; Yingju LIN ; Fuujen TSAI
Biomedical and Environmental Sciences 2011;24(3):284-290
OBJECTIVEGenistein, a major soy isoflavone metabolite (SIF), inactivates oxidation activity of bovine lactoperoxidase (LPO). Modification of the heme moiety of LPO by nitrogen-containing compounds has been shown to inactivate LPO. In contrast, SIF mediated inactivation of LPO does not involve a heme modification and the mechanism of SIF inhibition is poorly understood.
METHODSAfter inactivation of LPO by genistein in the presence of H(2)O(2), trypsin-digested LPO peptide fragments were collected and analyzed by MALDI-TOF-MS to characterize the chemical binding of genistein(s) to LPO.
RESULTSThe heme moiety of LPO was not modified by genistein. A covalent binding study showed that (3)H-genistein bound to LPO with a ratio of ∼12 to 1. After HPLC analysis and peak collection, trypsin-digested peptide fragments were analyzed by MALDI-TOF-MS. The 3H-genistein co-eluted peptide fragments (RT=24 min) were putatively identified as 199IVGYLDEEGVLDQNR214 with two bound genistein molecules or a genistein dimer (2 259 Da), 486TPDNIDIWIGGNAEPMVER504 with two bound genistein molecules or a genistein dimer (2 663 Da), and 161ARWLPAEYEDGLALPFGWTQR182 with three bound genistein molecules or a genistein trimer (3 060 Da). The fragment with a mass of 1 792 Da (RT=36 min) was identified as 132CDENSPYR139 with three genistein molecules or a genistein trimer.
CONCLUSIONSThe results suggest that LPO was inactivated by irreversible covalent binding of genistein or genistein polymers to particular peptide fragments constituting regions of the outward domain. No genistein interaction with the prosthetic heme moiety of LPO was observed.
Animals ; Cattle ; Enzyme Activation ; drug effects ; Genistein ; metabolism ; Hydrogen Peroxide ; pharmacology ; Isoflavones ; pharmacology ; Lactoperoxidase ; metabolism ; Placental Lactogen ; Protein Binding
10.Effects of genistein and zearalenone on proliferation of PEO4.
Zeng-li YU ; Li-shi ZHANG ; Qun-ying LI ; De-sheng WU
Chinese Journal of Preventive Medicine 2003;37(3):154-157
OBJECTIVEThe objective of this study was to investigate the estrogenic activity of genistein and zearalenone through their effects on the proliferative capacity of human ovarian PEO4.
METHODSEstrogen receptor-positive PEO4 cell was grown in DMEM medium containing 10% bovine serum. Five days before the addition of the test compounds, the cells were washed in phosphate-buffered saline, and the medium was substituted with a phenol red-free DMEM medium containing 5% dextran charcoal-stripped FBS. The respective test compound was added in fresh medium and the control cell received only the vehicle (ethanol). Cell proliferation was detected respectively by MTT assay, (3)H-TdR incorporation and flow cytometry.
RESULTSCompared with vehicle control, 96 x 10(-6) mol/L GS significantly inhibited PEO4 cell proliferation and DNA synthesis as measured by MTT and (3)H-TdR incorporation after treatment for 24 h. Alao, 32 x 10(-6) mol/L GS could exert inhibition on PEO4 cell growth as time extension to 48 h. 32 x 10(-6) mol/L approximately 96 x 10(-6) mol/L GS induced G(2)/M arrest. At low dose (< 8 x 10(-6) mol/L=, GS promoted proliferation in PEO4 cells. ZEA enhanced proliferation, promoted DNA synthesis and increased the S phase population in PEO4 cells.
CONCLUSIONSGenistein possess estrogenic activity and zearalenone have anti-estrogenic activity. They play different effects on the proliferation of human ovarian cancer cell. Genistein enhanced the proliferation of PEO4. Zearalenone inhibited its the proliferation. These results implied that genistein and zearalenone elicit different signal-transduction channel.
Antineoplastic Agents ; pharmacology ; Cell Division ; drug effects ; Estrogens, Non-Steroidal ; pharmacology ; Female ; Genistein ; pharmacology ; Humans ; Ovarian Neoplasms ; pathology ; Receptors, Estrogen ; metabolism ; Tumor Cells, Cultured ; Zearalenone ; pharmacology