Molecular mechanism of Spatholobi Caulis in treatment of ovarian cancer based on network pharmacology and experimental verification / 中国中药杂志

The present study explored the main active ingredients and the underlying mechanism of Spatholobi Caulisin the treatment of ovarian cancer(OC) by network pharmacology, molecular docking, and in vitro cell experiments. The active ingredients and their predicted targets(AITs) were first acquired online with the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP). Theoretical disease targets(DTs) were obtained through professional databases including GeneCards, OMIM, PharmGkb, TTD, and DrugBank. The common targets in the intersection of AITs and DTs were used for the construction of a "drug-ingredient-disease-target" network by Cytoscape 3.7.1. STRING database was used to construct a protein-protein interaction(PPI) network. R 4.0.5 was used for GO and KEGG functional enrichment analyses. Schr9 dinger Maestro was used to perform and optimize the molecular docking and virtual screening.Twenty-three active ingredients of Spatholobi Caulis were screened out, involving 75 OC targets and 178 signaling pathways.Network analysis revealed that Spatholobi Caulis presumedly exerted an anti-OC effect by acting on key protein targets such as GSK-3β, Bcl-2, and Bax. Molecular docking showed that GSK-3β possessed goodbinding activity to prunetin. In vitro cell experiments preliminarily verified the core targets and pathways of prunetin, the active ingredient of Spatholobi Caulis against human OC SKOV3 cells.CCK-8 assay was used to detect the cell proliferation, and flow cytometry was used to detect the effect of prunetin on apoptosis of human OC SKOV3 cells.The expression of prunetin targets and related regulatory proteins was detected by Western blot.In vitro cell experiments demonstrated that prunetindisplayed significant inhibitory effects on the proliferation of OC cells and could induce apoptosis of SKOV3 cells. Western blot showed that prunetin could induce SKOV3 cell apoptosis by inhibiting GSK-3β phosphorylation and regulating the expression of downstream Bcl-2 and Bax proteins. This study reveals the scientific nature of network pharmacology in the prediction and guidance of experimental design, confirming that prunetin can treat OC by blocking the GSK-3β/Bcl-2/Bax cell signal transduction pathway. The findings are expected to provide a basis for the investigation of the mechanism of Spatholobi Caulis in the treatment of OC.

Study on isoflavones isolated from stems of Wisteria sinensis / 中草药

Objective: To investigate the chemical constituents from the stems of Wisteria sinensis. Methods: The 14 chemical constituents were isolated and purified by silica gel, polyamide, Sephadex LH-20 column chromatography, and semi-preparative HPLC, and their chemical structures were identified by physico-chemical constants and spectral data. Results: Fourteen isoflavones were isolated from the methanol extract of the stems of W. sinensis and identified as tectorigenin-7-O-β-D-(6″-O-acetyl)-glucoside (1), 7,3',4'-trihydroxyisoflavanone (2), formononetin (3), afromosin (4), prunetin (5), biochanin A (6), gliricidin (7), 8-O-methylreyusin (8), tectoridin (9), genistin (10), sissotrin (11), ononin (12), kakkanin (13), and kushenol O (14). Conclusion: Compound 1 is a new compound named as tectoridin A, and compounds 2, 5, 7, 9-11, 13, and 14 are obtained from the genus Wisteria for the first time.

Effects of prunetin on the proteolytic activity, secretion and gene expression of MMP-3 in vitro and production of MMP-3 in vivo

We investigated whether prunetin affects the proteolytic activity, secretion, and gene expression of matrix metalloproteinase-3 (MMP-3) in primary cultured rabbit articular chondrocytes, as well as in vivo production of MMP-3 in the rat knee joint to evaluate the potential chondroprotective eff ect of prunetin. Rabbit articular chondrocytes were cultured in a monolayer, and reverse transcription-polymerase chain reaction (RT-PCR) was used to measure interleukin-1beta (IL-1beta)-induced expression of MMP-3, MMP-1, MMP-13, a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4), and ADAMTS-5. In rabbit articular chondrocytes, the effects of prunetin on IL-1beta-induced secretion and proteolytic activity of MMP-3 were investigated using western blot analysis and casein zymography, respectively. The eff ect of prunetin on MMP-3 protein production was also examined in vivo. The results were as follows: (1) prunetin inhibited the gene expression of MMP-3, MMP-1, MMP-13, ADAMTS-4, and ADAMTS-5; (2) prunetin inhibited the secretion and proteolytic activity of MMP-3; (3) prunetin suppressed the production of MMP-3 protein in vivo. These results suggest that prunetin can regulate the gene expression, secretion, and proteolytic activity of MMP-3, by directly acting on articular chondrocytes.

Isoflavones from flower buds of Rosa rugosa and their cytotoxicity / 中草药

Objective: To study the chemical constituents from the flower buds of Rosa rugosa. Methods: The chemical constituents from the flower buds of R. rugosa were isolated by silica gel, MCI-gel resin, Sephadex LH-20 column chromatography and high performance liquid chromatography (HPLC) methods. Their structures were elucidated by spectroscopic methods and physicochemical properties. Results: Three isoflavones, 6,8-dihydroxy-4',7-dimethoxyisoflavone (1), prunetin (2), and pratensein (3) were isolated from the flower buds of R. rugosa. Conclusion: Compound 1 is a new compound named rosa isoflavone. Compounds 2 and 3 are isolated from R. rugosa for the first time. Compound 1 displays the stronger cytotoxicity against A549 and PC3 cells with IC50 values of 2.6 and 3.2 μmol/L, respectively.

Chemical constituents from roots of Astragalus englerianus and their anti-oxidative activities / 中草药

Objective: To study the chemical constituents from the roots of Astragalus englerianus, and to determine their anti-oxidative activities. Methods: The compounds were isolated and purified by silica gel, RP18, and Sephadex LH-20 column chromatography, then their structures were elucidated on the basis of spectral data and physicochemical properties, and anti-oxidative activities were tested by DPPH method. Results: Twenty-nine compounds were obtained from the ethyl acetate fraction of the methanol extract from the roots of A. englerianus and identified as isoliquiritigenin (1), 4'-hydroxy-2, 4-dimethoxychalcone (2), xenognosin (3), formononetin (4), calycosin (5), prunetin (6), (3R)-vestitol (7), liquiritigenin (8), (6aR, 11aR)-medicarpin-3-O-β-D-glucopyranoside (9), olean-12-en-3β, 22β, 24-triol (10), friedelin (11), β-sitosterol (12), stigmasterol (13), 7β-hydroxysitosterol (14), 7-oxositosterol (15), 3β-sitosteryl (9'Z)-9'-heptadecenoate (16), stigmast-4-en-3-one (17), 5α, 8α-epidioxy-ergosta-6, 9, 22E-trien-3β-ol (18), 5α, 8α-epidioxy-ergosta-6, 22E-dien-3β-ol (19), D-2-O-methylinositol (20), octacosanol (21), methyl stearate (22), eicosanoic acid (23), heneicosanoic acid (24), oleic acid (25), linoleic acid (26), α-linolenic acid (27), tripalmitin (28), and trilinolein (29). The ethyl acetate soluble portion, compounds 1 and 3 showed DPPH free radical scavenging activity with IC50 values of (66.0 ± 1.8), (64.3 ± 0.4), and (57.1 ± 1.2) μg/mL, respectively. Conclusion: This is the first report on the compounds 2, 3, 6, 10, 11, 14-22, and 28 from the plants of Astragalus Linn., and all the compounds are obtained from A. englerianus for the first time. A. englerianus is found to possess the potent anti-oxidative activity.

Chemical constituents from leaves of Cajanus cajan / 中草药

Objective: To study the chemical compounds from the leaves of Cajanus cajan. Methods: Normal phase silica gel, medium-pressure ODS, MCI, and Sephadex LH-20 gel column chromatographies were used to isolate and purify the constituents. And their structures were identified by their physical properties, nuclear magnetic resonance spectrum, mass spectrometry, etc. Results: Twenty-three compounds were separated from the alcohol extract in the leaves of C. cajan. They were cajanine (1), longistyline A (2), longistyline C (3), cajanolactone A (4), pinostrobin (5), orientin (6), isovitexin (7), vitexin (8), cajanol (9), cajanin (10), prunetin (11), pratensein (12), (2R, 3R)-2, 3-dihydro-5-7, 4'-dimethoxyflavone (13), ethyl 10', 16'-dihydroxy hexadecanoate (14), vanillic acid (15), ethyl heptadecanoate (16), 2-O-quebrachitol (17), 2, 3, 4-trihydroxy-isovaleric acid (18), stigmasterol (19), betulinic acid (20), heptadecanoic acid (21), β-sitosterol (22), and β-daucosterol (23). Conclusion: Compounds 10-19 are obtained from this plant for the first time.

Effect of Prunetin on TNF-alpha-Induced MUC5AC Mucin Gene Expression, Production, Degradation of IkappaB and Translocation of NF-kappaB p65 in Human Airway Epithelial Cells / 결핵및호흡기질환

BACKGROUND: We investigated whether prunetin significantly affects tumor necrosis factor-alpha (TNF-alpha)-induced MUC5AC mucin gene expression, production, inhibitory kappa B (IkappaB) degradation and nuclear factor kappa B (NF-kappaB) p65 translocation in human airway epithelial cells. METHODS: Confluent NCI-H292 cells were pretreated with prunetin for 30 minutes and then stimulated with TNF-alpha for 24 hours or the indicated periods. MUC5AC mucin gene expression and mucin protein production were measured by reverse transcription polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. The effect of prunetin on TNF-alpha-induced degradation of IkappaB and translocation of NF-kappaB p65 was investigated by western blot analysis. RESULTS: We found that incubation of NCI-H292 cells with prunetin significantly inhibited mucin production and down-regulated the MUC5AC gene expression induced by TNF-alpha. Prunetin inhibited TNF-alpha-induced degradation of IkappaB and translocation of NF-kappaB p65. CONCLUSION: This result suggests that prunetin inhibits the NF-kappaB signaling pathway, which may explain its role in the inhibition of MUC5AC mucin gene expression and production regulated by the NF-kappaB signaling pathway.

Chemical Constituents from Roots of Flemingia philippinensis / 中草药·英文版

ObjectiveTo study the chemical constituents from the roots of Flemingia philippinensis.MethodsThe chemical constituents were isolated and purified by combination of silica gel column,Sephadex LH-20,polyamide,and ODS column chromatography.The structures of the isolated compounds were identified by means of spectral data.ResultsTen compounds were isolated from F.philippinensis and identified as isoderrone (1),dalparvin A (2),prunetin (3),7,3'-dihydroxy-5,4',5'-trimethoxyisoflavone (4),pratensein-7-O-β-D-glucoside (5),sissotrin (6),sophororicoside (7),formononetin (8),orobol (9),and biochanin A (10).ConclusionCompounds 1-6 are obtained from this plant for the first time.

Regulation of Tumor Necrosis Factor-alpha-induced Airway Mucin Production and Gene Expression by Carbenoxolone, Prunetin, and Silibinin / 결핵및호흡기질환

BACKGROUND: In this study, we tried to investigate whether carbenoxolone, prunetin, and silibinin affect tumor necrosis factor (TNF)-alpha-induced MUC5AC mucin production and gene expression from human airway epithelial cells. METHODS: Confluent NCI-H292 cells were pretreated with each agent (carbenoxolone, prunetin, and silibinin) for 30 min and then stimulated with TNF-alpha for 24 hours. The MUC5AC mucin gene expression and mucin protein production were measured by reverse transcription-polymerase chain reaction and enzyme linked immunosorbent assay, respectively. RESULTS: Carbenoxolone, prunetin and silibinin inhibited the production of MUC5AC mucin protein induced by TNF-alpha; the 3 compounds also inhibited the expression of MUC5AC mucin gene induced by TNF-alpha. CONCLUSION: This result suggests that carbenoxolone, prunetin and silibinin can inhibit mucin gene expression and production of mucin protein induced by TNF-alpha, by directly acting on airway epithelial cells.