Bo-Yang Yu

Anti-inflammatory Effects of Aqueous Extract from Radix Liriope muscari and Its Major Active Fraction and Component

Aim To investigate the anti-inflammatory activities of the aqueous extract from Liriope muscari (Decne.) Bailey (Lm-a), its crude saponin fraction (Lm-s) and one major component (Lm-3), and to provide some pharmacological evidence for its clinical use in inflammatory diseases.

Advances in the pharmacological activities and mechanisms of diosgenin.

Diosgenin, a well-known steroid sapogenin derived from plants, has been used as a starting material for production of steroidal hormones. The present review will summarize published literature concerning pharmacological potential of diosgenin, and the underlying mechanisms of actions. Diosgenin has shown a vast range of pharmacological activities in preclinical studies. It exhibits anticancer, cardiovascular protective, anti-diabetes, neuroprotective, immunomodulatory, estrogenic, and skin protective effects, mainly by inducing apoptosis, suppressing malignant transformation, decreasing oxidative stress, preventing inflammatory events, promoting cellular differentiation/proliferation, and regulating T-cell immune response, etc. It interferes with cell death pathways and their regulators to induce apoptosis. Diosgenin antagonizes tumor metastasis by modulating epithelial-mesenchymal transition and actin cytoskeleton to change cellular motility, suppressing degradation of matrix barrier, and inhibiting angiogenesis. Additionally, diosgenin improves antioxidant status and inhibits lipid peroxidation. Its anti-inflammatory activity is through inhibiting production of pro-inflammatory cytokines, enzymes and adhesion molecules. Furthermore, diosgenin drives cellular growth/differentiation through the estrogen receptor (ER) cascade and transcriptional factor PPARγ. In summary, these mechanistic studies provide a basis for further development of this compound for pharmacotherapy of various diseases.

Analysis of Flavonoids and Phenolic Acids in Iris tectorum by HPLC-DAD-ESI-MS~n

Abstract Aim To develop high performance liquid chromatography combined with photodiode-array detection and electrospray ionization multiple-stage mass spectrometry (HPLC-DAD-ESI-MS n ) for the analysis and identification of flavonoids and phenolic acids in the rhizome of Iris tectorum Maxim‥ Method The structures of flavonoids and phenolic acids were identified by chromatographic retention times, UV spectra as well as ESI-MS n spectra. Results Ten isoflavones were identified as tectorigenin-7- O -β-glucosyl-4′- O -β-glucoside (3), tectoridin (5) , iristectorin B (6) , iristectorin A (7) , iridin (8) , genistein (11) , tectorigenin (12) , iristectorigenin A (14) , iristectorigenin B (16), i and rigenin (17) . Two flavanones, one flavonol and one flavanonol were tentatively identified as hesperetin (9) , 5, 7, 3′-trihydroxy-6, 4′-dimethoxyflavanone (10), rhamnocitrin (13) and dihydrokaempferide (15), respectively. The three phenolic acids were tectoruside (1) , androsin (2) and apocynin (4) . Conclusion The developed simple and rapid method is useful to rapidly identify the bioactive constituents in the rhizome of Iris tectorum . Two flavanones, hesperetin (9) and 5,7,3′-trihydroxy-6, 4′-dimethoxyflavanone (10) were identified from this species for the first time.

Simultaneous Determination of 26 Pesticide Residues in 5 Chinese Medicinal Materials Using Solid-phase Extraction and GC-ECD Method

AIM: To develop an analytical method for the simultaneous determination of 26 pesticide residues including 21 organochlorines and 5 pyrethroids in 5 crude Chinese medicinal materials by gas chromatography-electron capture detector (GC-ECD) method. METHODS: Florisil solid-phase extraction (SPE) was used for the clean-up of samples and GC-ECD for the determination. The separation of the target pesticides was carried out on HP-5 quartz capillary column in an optimized oven program with nitrogen as a carrier at a constant flow of 1.0 mL•min^(-1) and the injection inlet valve was set at 260℃ and ECD at 330℃. RESULTS: The florisil solid-phase extraction was effective for the clean-up of the samples. For 26 pesticides, LODs and LOQs of the developed method were 0.386-2.500 ng•g^(-1) and 0.879-6.250 ng•g^(-1) respectively, and the average recoveries were found to be 69.61%-112.80% (RSD=2.43%-15.18%, n=9). CONCLUSION: The developed method could be used to simultaneously determine the 26 pesticide residues in 5 Chinese medicinal materials, and it could also be potential in the determination of other similar Chinese medicinal materials.

DT-13, a saponin of dwarf lilyturf tuber, exhibits anti-cancer activity by down-regulating C-C chemokine receptor type 5 and vascular endothelial growth factor in MDA-MB-435 cells

AIM To investigate the anticancer activity of DT-13 under normoxia and determine the underlying mechanisms of action. METHODS MDA-MB-435 cell proliferation, migration, and adhesion were performed to assess the anticancer activity of DT-13, a saponin from Ophiopogon japonicus, in vitro. In addition, the effects of DT-13 on tumor growth and metastasis in vivo were evaluated by orthotopic implantation of MDA-MB-435 cells into nude mice; mRNA levels of vascular endothelial growth factor (VEGF), C-C chemokine receptor type 5 (CCR5) and hypoxia-inducible factor 1α (HIF-1α) were evaluated by real-time quantitative PCR; and CCR5 protein levels were detected by Western blot assay. RESULTS At 0.01 to 1 μmol·L(-1), DT-13 inhibited MDA-MB-435 cell proliferation, migration, and adhesion significantly in vitro. DT-13 reduced VEGF and CCR5 mRNAs, and decreased CCR5 protein expression by down-regulating HIF-1α. In addition, DT-13 inhibited MDA-MB-435 cell lung metastasis, and restricted tumor growth slightly in vivo. CONCLUSION DT-13 inhibited MDA-MB-435 cell proliferation, adhesion, and migration in vitro, and lung metastasis in vivo by reducing VEGF, CCR5, and HIF-1α expression.

Homoisoflavonoids from Ophiopogon japonicus and Its Oxygen Free Radicals (OFRs) Scavenging Effects

ABSTRACT AIM To investigate the OFRs scavenging activities of homoisoflavonoids from the fibrous root of Ophiopogon japonicus . METHODS The scavenging effect of homoisoflavonoids isolated from O. japonicus extracts was examined on superoxide anion (O 2 ·− ), hydroxyl radical (·OH), and hydrogen peroxide (H 2 O 2 ) by chemiluminescence. RESULTS Ten homoisoflavonoids were obtained from O. japonicus extracts. They were identified as methylophiopogonanone A ( 1 ), methylophiopogonanone B ( 2 ), ophiopogonanone A ( 3 ), ophiopogonanone E ( 4 ), 5, 7-dihydroxy-6, 8-dimethyl-3-(4′-hydroxy-3′, 5′-methoxybenzyl) chroman-4-one ( 5 ), methylophiopogonone A ( 6 ), methylophiopogonone B ( 7 ), desmethylisoophiopogonone B ( 8 ), 5, 7, 2′-trihydroxy-8-methyl-3-(3′, 4′-methylenedioxybenzyl) chromone ( 9 ) and 5, 7, 2′-trihydroxy-6, 8-dimethyl-3-(3′, 4′-methylenedioxybenzyl) chromone ( 10 ). CONCLUSION Compounds 8 and 9 were firstly isolated from O. japonicus . The majority of homoisoflavonoids could scavenge ·OH and H 2 O 2 in vitro to a certain extent, and their bioactivities should be related with the respective structures.

Structural characterization and identification of major constituents in Radix Scrophulariae by HPLC coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry

AIM To analyze the major constituents in Radix Scrophulariae (Scrophularia ningpoensis). METHOD Radix Scrophulariae was analyzed by HPLC coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (ESI-Q-TOF MS/MS). Compounds were separated by HPLC using a C18 column and gradient elution of acetonitrile and 0.1 % (V/V) acetic acid-water. Negative ion mode was employed. RESULTS A total of thirty-six compounds, including fourteen iridoid glycosides, nineteen phenylpropanoid glycosides, and three organic acids, were identified from Radix Scrophulariae based on the accurate mass measurement of precursor and product ions. Twenty-one of the constituents were identified by comparing their retention times (tR) and ESI-MS/MS data with those of reference standards and/or previous publications, while another fifteen compounds were tentatively identified or deduced according to their Q-TOF MS/MS data which afforded sufficient structural information. CONCLUSION It is believed that this study is useful for the identification of constituents in Radix Scrophulariae, as well as related plants and complex prescriptions.

The saponin DT-13 inhibits gastric cancer cell migration through down-regulation of CCR5-CCL5 axis.

AIM To investigate the effect of DT-13 on gastric cancer cell migration, and to explore the possible mechanisms underlying the anti-metastasis activity of DT-13. METHODS Growth inhibition of DT-13 was analyzed by the MTT assay. Cell migration was measured by the scratch-wound assay and transwell double chamber assay. To investigate the possible mechanisms underlying the anti-metastasis activity of DT-13, chemokine receptors that are involved in cancer metastasis (CCR2, CCR5, CCR7, CXCR4, and CXCR6) were detected by conventional PCR. The effect of DT-13 on CCR5 and CXCR4 expression was further evaluated by quantitative PCR and Western blot, respectively. The secretion of CCL5 (ligand of CCR5) and SDF-1 (ligand of CXCR4) were detected by enzyme-linked immunosorbent assay (ELISA). RESULTS DT-13 inhibited BGC-823 and HGC-27 cell growth in a dose dependent manner, and the estimated IC50 value for 24 h treatment was 23.5 ± 5.1 μmol·L(-1) for BGC-823 cells and 35.6 ± 7.6 μmol·L(-1) for HGC-27 cells. DT-13 also significantly decreased gastric cancer cell migration. DT-13 significantly decreased the gene expression of CCR5 in both BGC-823 and HGC-27 gastric cancer cells, and moderately reduced the expression of CXCR4. Similar to the results of gene expression, significant down-regulation of CCR5 protein was observed, but CXCR4 protein levels were much less affected. CCL5 secretion, but not SDF-1 production, was inhibited by DT-13. CONCLUSION DT-13 inhibited gastric cancer cell migration by down-regulation of the CCR5-CCL5 axis.

Diosgenin reduces leukocytes adhesion and migration linked with inhibition of intercellular adhesion molecule-1 expression and NF-κB p65 activation in endothelial cells

Abstract Aim To investigate the anti-inflammatory activity and possible mechanism of diosgenin, and to provide some further pharmacological evidence for its clinical applications. Methods Anti-inflammatory activities of diosgenin were evaluated by zymosan A-evoked peritoneal leukocytes migration in mice and the adhesion of human pro-myelocytic leukemia cell strain (HL-60) to human umbilical vein endothelial cell line (ECV304) cells induced by tumor necrosis factor (TNF-α1;). Furthermore, the effects of diosgenin on TNF-α1;-induced intercellular adhesion molecule-1 (ICAM-1) expression were also investigated by reverse transcription-PCR (RT-PCR) and flow cytometric analysis, and nuclear factor-kappaB (NF-αB)/p65 translocation and phosphorylation were determined by Western blot. Results Diosgenin significantly inhibited peritoneal leukocytes migration induced by zymosan A in mice when given orally once at doses of 1 and 3 mg·kg−1. Pretreatment ECV304 cells with diosgenin at concentrations of 0.1 and 1 μmol·L−1 for 5 h remarkably decreased TNF-α1;-stimulated adhesion of HL-60 to ECV304 cells in vitro, with no effect on viability of ECV304 cells. Furthermore, diosgenin inhibited TNF-α-induced overexpression of ICAM-1 both at the mRNA and protein levels, and suppressed nuclear p65 accumulation and p65 activation in ECV304 cells. Conclusion These results suggested that diosgenin significantly inhibited leukocytes migration and adhesion, partly due to the down-regulation of ICAM-1 expression through NF-κB pathway.

Network pharmacology-based prediction and verification of the molecular targets and pathways for schisandrin against cerebrovascular disease.

AIM To illuminate the molecular targets for schisandrin against cerebrovascular disease based on the combined methods of network pharmacology prediction and experimental verification. METHOD A protein database was established through constructing the drug-protein network from literature mining data. The protein-protein network was built through an in-depth exploration of the relationships between the proteins. The computational platform was implemented to predict and extract the sensitive sub-network with significant P-values from the protein-protein network. Then the key targets and pathways were identified from the sensitive sub-network. The most related targets and pathways were also confirmed in hydrogen peroxide (H2O2)-induced PC12 cells by Western blotting. RESULTS Twelve differentially expressed proteins (gene names: NFKB1, RELA, TNFSF10, MAPK1, CHUK, CASP8, PIGS2, MAPK14, CREB1, IFNG, APP, and BCL2) were confirmed as the central nodes of the interaction network (45 nodes, 93 edges). The NF-κB signaling pathway was suggested as the most related pathway of schisandrin for cerebrovascular disease. Furthermore, schisandrin was found to suppress the expression and phosphorylation of IKKα, as well as p50 and p65 induced by H2O2 in PC12 cells by Western blotting. CONCLUSION The computational platform that integrates literature mining data, protein-protein interactions, sensitive sub-network, and pathway results in identification of the NF-κB signaling pathway as the key targets and pathways for schisandrin.