[Show abstract][Hide abstract] ABSTRACT: Novel agents need to be developed to overcome the limitations of the current melanoma therapeutics. Atractylenolide I (AT-I) is a sesquiterpene compound isolated from atractylodis macrocephalae rhizoma. Previous findings demonstrated that AT-I exhibited cytotoxic action in melanoma cells. However, the molecular mechanisms of AT‑1's anti-melanoma properties remain to be elucidated. In the present study, the cell cycle-arrest and apoptosis-promoting effects as well as the ERK/GSK3β signaling-related mechanism of action of AT-I were examined. B16 melanoma cells were treated with various concentrations of AT-1 (50, 75 and 100 µM) for 48 or 72 h. Cell cycle and apoptosis were analyzed by flow cytometry. Protein expression levels were detected by western blot analysis. AT-I treatment induced G1 phase arrest, which was accompanied by increased p21 and decreased CDK2 protein expression levels. Apoptosis was observed after AT-I treatment for 72 h, which was accompanied by activated caspase‑3 and ‑8. AT-I treatment significantly decreased phospho-ERK, phospho-GSK3β, c-Jun and increased p53 protein expression levels. Lithium chloride (LiCl, 5 mM), a GSK3β inhibitor, treatment alone did not increase the apoptosis of B16 cells, while pretreatment with LiCl markedly reversed AT-I-induced apoptosis. Additionally, AT-I-induced G1 phase arrest was partially reversed by LiCl pretreatment. In conclusion, ERK/GSK3β signaling was involved in the apoptotic and G1 phase arrest effects of AT-I in melanoma cells.
[Show abstract][Hide abstract] ABSTRACT: Two new rare α-pyrone (=2H-pyran-2-one) derivatives, rhodanthpyrones A and B (1 and 2, resp.), together with fourteen known compounds, 3-16, were isolated from the whole plants of Gentiana rhodantha. The structures of these compounds were elucidated by spectroscopic analyses. This is the first report on the occurrence of α-pyrone derivatives in the genus Gentiana.
[Show abstract][Hide abstract] ABSTRACT: Norisoboldine (NOR), the primary isoquinoline alkaloid constituent of the root of Lindera aggregata, has previously been demonstrated to attenuate osteoclast (OC) differentiation. Accumulative evidence has shown that aryl hydrocarbon receptor (AhR) plays an important role in regulating the differentiation of various cells, and multiple isoquinoline alkaloids can modulate AhR. In the present study, we explored the role of NOR in the AhR signaling pathway. These data showed that the combination of AhR antagonist resveratrol (Res) or α-naphthoflavone (α-NF) nearly reversed the inhibition of OC differentiation through NOR. NOR could stably bind to AhR, up-regulate the nuclear translocation of AhR, and enhance the accumulation of the AhR-ARNT complex, AhR-mediated reporter gene activity and CYP1A1 expression in RAW 264.7 cells, suggesting that NOR might be an agonist of AhR. Moreover, NOR inhibited the nuclear translocation of NF-κB-p65, resulting in the evident accumulation of the AhR-NF-κB-p65 complex, which could be markedly inhibited through either Res or α-NF. Although NOR only slightly affected the expression of HIF-1α, NOR markedly reduced VEGF mRNA expression and ARNT-HIF-1α complex accumulation. In vivo studies indicated that NOR decreased the number of OCs and ameliorated the bone erosion in the joints of rats with collagen-induced arthritis, accompanied by the up-regulation of CYP1A1 and the down-regulation of VEGF mRNA expression in the synovium of rats. A combination of α-NF nearly completely reversed the effects of NOR. In conclusion, NOR attenuated OC differentiation and bone erosion through the activation of AhR and the subsequent inhibition of both NF-κB and HIF pathways.
International journal of biological sciences 01/2015; 11(9):1113-26. DOI:10.7150/ijbs.12152 · 4.37 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Eleven new sesquiterpenoids, wenyujinins A-K (1-11), and a new monoterpenoid, wenyujinin L (12), were isolated from the rhizomes of Curcuma wenyujin. Their structures and relative configurations were elucidated using 1D and 2D NMR, X-ray crystallographic analysis, and HRESIMS data. The absolute configurations of 1, 2, 3, 4, 6, 8, 9, and 10 were determined by comparison of the experimental and calculated ECD spectra. The absolute configuration of 5 was determined from the ECD data of the [Rh2(OCOCF3)4] complex, whereas those of 7 and 12 were determined from the ECD spectra of the compounds alone. Compounds 7 and 7a strongly inhibited the induction of NO production by LPS, with IC50 values of 7.6 and 8.5 μM, respectively. Compounds 6 and 10 moderately inhibited NO production with IC50 values of 47.7 and 48.6 μM, respectively.
Journal of Natural Products 10/2014; DOI:10.1021/np400984c · 3.95 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Norisoboldine (NOR) is a benzylisoquinoline alkaloid isolated from Radix Linderae, a traditional Chinese medicine. Our previous studies have demonstrated that it produces anti-inflammatory and anti-rheumatoid arthritis effects.
The present study was undertaken to explore the analgesic effects of NOR and its potential mechanism in the formalin test and the acetic acid writhing test.
Oral administration of NOR dose dependently attenuated the formalin-induced pain responses in the second phase, and reduced formalin-induced paw oedema. It also diminished acetic acid-induced writhing responses but had no effect on acute thermal pain in the hotplate test. The mechanistic studies suggested that the adenosine system, but not the opioid receptor system, is involved in NOR-induced antinociception. Naloxone, a non-selective opioid receptor antagonist, had no effect on NOR-induced analgesic action. However, caffeine (a non-selective adenosine receptor antagonist) completely reversed the analgesic effect of NOR in formalin-induced nociceptive responses in the second phase, and 8-cyclopentyl-1, 3-dipropylxanthine (DPCPX, a selective adenosine A1 receptor antagonist) completely inhibited NOR-induced analgesia in both formalin-induced nociceptive responses and acetic acid-induced writhing responses. In addition, NOR reduced formalin-induced activation of extracellular signal-regulated kinase and calcium/calmodulin-dependent protein kinase II in the spinal cord, which is also blocked by DPCPX. Furthermore, NOR decreased forskolin-evoked cyclic adenosine monophosphate levels in mouse spinal cord neuronal cultures through the adenosine A1 receptor.
Our data demonstrate that NOR produces the analgesic effect in inflammatory pain by a mechanism related to the adenosine system.
European journal of pain (London, England) 08/2014; 18(7). DOI:10.1002/j.1532-2149.2013.00439.x · 3.22 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Abietane derivatives, bungnates A, B, 15-dehydrocyrtophyllone A and 15-dehydro-17-hydroxycyrtophyl-lone A, and two phenylethanoid glycosides, bunginoside A and 3 '',4 ''-di-O-acetylmartynoside, together with nine known abietane derivatives and fourteen known phenylethanoid glycosides, were isolated from dried roots of Clerodendrum bungei. Their structures were determined on the basis of detailed spectroscopic analyses and acidic hydrolysis. The absolute configuration of bunginoside A was established from analysis of CD data. Selected compounds were evaluated for inhibitory effects against angiotensin converting enzyme (ACE) and alpha-glucosidase. 15-Dehydrocyrtophyllone A showed an ACE inhibitory effect, and verbascoside, leucosceptoside A and isoacteoside exhibited strong inhibitory capacity against alpha-glucosidase. (c) 2014 Elsevier Ltd. All rights reserved.
[Show abstract][Hide abstract] ABSTRACT: Six new diterpenoids, 4-epi-7α-O-acetylscoparic acid A (1), 7α-hydroxyscopadiol (2), 7α-O-acetyl-8,17β-epoxyscoparic acid A (3), neo-dulcinol (4), dulcinodal-13-one (5), and 4-epi-7α-hydroxydulcinodal-13-one (6), and a new flavonoid, dillenetin 3-O-(6″-O-p-coumaroyl)-β-d-glucopyranoside (10), along with 12 known compounds, were isolated from the aerial parts of Scoparia dulcis. The 7S absolute configuration of the new diterpenoids 1-4 and 6 was deduced by comparing their NOESY spectra with that of a known compound, (7S)-4-epi-7-hydroxyscoparic acid A (7), which was determined by the modified Mosher's method. The flavonoids scutellarein (11), hispidulin (12), apigenin (15), and luteolin (16) and the terpenoids 4-epi-scopadulcic acid B (9) and betulinic acid (19) showed more potent α-glucosidase inhibitory effects (with IC50 values in the range 13.7-132.5 μM) than the positive control, acarbose. In addition, compounds 1, 11, 12, 15, 16, and acerosin (17) exhibited peroxisome proliferator-activated receptor gamma (PPAR-γ) agonistic activity, with EC50 values ranging from 0.9 to 24.9 μM.
[Show abstract][Hide abstract] ABSTRACT: A new triglycoside flavone, luteolin-4'-O-α-l-rhamnopyranosyl-(1 → 6)-β-d-glucopyranosyl-(1 → 3)-β-d-glucopyranoside (1) was isolated from the leaves of Ficus ischnopoda, together with five known flavonoids (2-6) and seven known lignans (7-13). Their structures were elucidated on the basis of extensive spectroscopic analysis and comparison with literature data. Compounds 1-13 were reported in this plant for the first time. Compounds 1 and 3-6 exhibited significant anti-HSV-1 activity in vitro.
Natural product research 04/2014; 28(14). DOI:10.1080/14786419.2014.904307 · 1.23 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Two new pyrrolizidine alkaloids, namely, neocroalbidine and neocroalbidinone, were isolated from the herbs of Crotalaria albida. Their structures were established using a combination of NMR spectroscopy and mass spectrometry techniques. The absolute stereochemistry of the two alkaloids presented herein was determined by single-crystal X-ray diffraction analysis.
[Show abstract][Hide abstract] ABSTRACT: Four new compounds, impecylone (1), deacetylimpecyloside (2), seguinoside K 4-methylether (3) and impecylenolide (4), were isolated from Imperata cylindrica along with two known compounds, impecyloside (5) and seguinoside K (6). Their structures were elucidated mainly by spectroscopic analyses including 1D- and 2D-NMR techniques, and the absolute configuration of 1 was confirmed by X-ray diffraction analysis. In calcium assay, the result indicated that compounds 1, 2, 4 and 5 cannot obviously inhibit the calcium peak value compared with the negative control, and suggested that the four compounds could not have anti-inflammatory activity.
Journal of Natural Medicines 07/2013; DOI:10.1007/s11418-013-0793-9 · 1.45 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Norisoboldine (NOR) is the main alkaloid constituent in the dry root of Lindera aggregata (Sims) Kosterm. (L. strychnifolia Vill.). As reported previously, orally administered NOR displayed a robust inhibition of joint bone destruction present in both mouse collagen-induced arthritis and rat adjuvant-induced arthritis with lower efficacious doses than that required for ameliorating systemic inflammation. This attracted us to assess the effects of NOR on differentiation and function of osteoclasts, primary effector cells for inflammatory bone destruction, to get insight into its anti-rheumatoid arthritis mechanisms.
Both RAW264.7 cells and mouse bone marrow-derived macrophages (BMMs) were stimulated with RANKL (100 ng/mL) to establish osteoclast differentiation models. ELISA, RT-PCR, gelatin zymography, western blotting, immunoprecipitation and EMSA were used to reveal related signalling pathways. NOR (10 and 30 µM), without significant cytotoxicity, showed significant reduction of the number of osteoclasts and the resorption pit areas, and it targeted osteoclast differentiation at the early stage. In conjunction with the anti-resorption effect of NOR, mRNA levels of cathepsin K and MMP-9 were decreased, and the activity of MMP-9 was attenuated. Furthermore, our mechanistic studies indicated that NOR obviously suppressed the ubiquitination of TRAF6, the accumulation of TRAF6-TAK1 complexes and the activation of ERK and p38 MAPK, and reduced the nuclear translocation of NF-κB-p65 and DNA-binding activity of NF-κB. However, NOR had little effect on expressions of TRAF6 or the phosphorylation and degradation of IκBα. Moreover, NOR markedly inhibited expressions of transcription factor NFATc1, but not c-Fos. Intriguingly, the subsequent nuclear translocations of c-Fos and NFATc1 were substantially down-regulated. Hence, we demonstrated for the first time that preventing the differentiation and function of osteoclasts at the early stage was an important anti-bone destruction mechanism of NOR, which might be attributed to inhibition of ubiquitination of TRAF6, the accumulation of TRAF6-TAK1 complexes and the activation of MAPKs/NF-κB/c-Fos/NFATc1 pathways.
PLoS ONE 03/2013; 8(3):e59171. DOI:10.1371/journal.pone.0059171 · 3.23 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Aim:
To explore the effects of norisoboldine (NOR), a major isoquinoline alkaloid in Radix Linderae, on joint destruction in rats with adjuvant-induced arthritis (AIA) and its underlying mechanisms.
AIA was induced in adult male SD rats by intradermal injection of Mycobacterium butyricum in Freund's complete adjuvant at the base of the right hind paw and tail. From d 14 after immunization, the rats were orally given NOR (7.5, 15, or 30 mg/kg) or dexamethasone (0.5 mg/kg) daily for 10 consecutive days. Joint destruction was evaluated with radiological scanning and H&E staining. Fibroblast-like synoviocytes (FLS) were prepared from fresh synovial tissues in the AIA rats. The expression of related proteins and mRNAs were detected by ELISA, Western blotting and RT-PCR.
In AIA rats, NOR (15 and 30 mg/kg) significantly decreased the swelling of paws and arthritis index scores, and elevated the mean body weight. NOR (30 mg/kg) prevented both the infiltration of inflammatory cells and destruction of bone and cartilage in joints. However, NOR (15 mg/kg) only suppressed the destruction of bone and cartilage, but did not obviously ameliorate synovial inflammation. NOR (15 and 30 mg/kg) significantly decreased the serum levels of receptor activator of nuclear factor κB ligand (RANKL), IL-6, PGE2, and MMP-13, but not the osteoprotegerin and MMP-1 levels. The mRNA levels of RANKL, IL-6, COX-2, and MMP-13 in synovium were also suppressed. Dexamethasone produced similar effects in AIA rats as NOR did, but without elevating the mean body weight. In the cultured FLS, treatment with NOR (10 and 30 mmol/L) significantly decreased the secretion of RANKL, IL-6, PGE2, and MMP-13 proteins. Furthermore, the treatment selectively prevented the activation of MAPKs, AKT and transcription factor AP-1 component c-Jun, but not the recruitment of TRAF6 or the activation of JAK2/STAT3. Treatment of the cultured FLS with the specific inhibitors of p38, ERK, AKT, and AP-1 significantly decreased the secretion of RANKL, IL-6, PGE2, and MMP-13 proteins.
NOR can alleviate joint destruction in AIA rats by reducing RANKL, IL-6, PGE2, and MMP-13 expression via the p38/ERK/AKT/AP-1 pathway.
[Show abstract][Hide abstract] ABSTRACT: AimTo study the chemical constituents of the rhizomes of Imperata cylindrica.Methods
The compounds were isolated by silica gel and Sephadex LH-20 column chromatography. Their structures were elucidated mainly by spectroscopic analyses including IR, HR-ESI-MS, 1D- and 2D-NMR.ResultsTwo new chromones, 8-hydroxy-2-(2-phenylethyl)chromone (1) and 2-(2-phenylethyl) chromone-8-O-β-D-glucopyranoside (2), and a new flavone glycoside, 4′-methoxyflavone-6-O-β-D-glucopyranoside (5), along with five known compounds, flidersiachromone (3), 5-hydroxy-2-(2-phenylethyl) chromone (4), flavone (6), 4′-hydroxy-5-methoxyflavone (7) and 5-hydroxyflavone (8) were isolated from Imperata cylindrica.Conclusion
Compounds 1, 2 and 5 were new compounds.
Chinese Journal of Natural Medicines 01/2013; 11(1):77–80. DOI:10.1016/S1875-5364(13)60012-6
[Show abstract][Hide abstract] ABSTRACT: To investigate the chemical constituents of Verbena officinalis L., five triterpenoid constituents were isolated and elucidated as 3α,19,23-trihydroxyurs-12-en-28-oic acid, namely, 4-epi-barbinervic acid (1), 2α,3β-dihydroxyurs-12-en-28-oic acid (2), 3α,24-dihydroxyurs-12-en- 28-oic acid (3), 3α,24-dihydroxy-olean-12-en-28-oic acid (4), ursolic acid (5), using spectroscopic methods and comparing with published data. Compounds 2 and 4 were obtained from V. officinalis for the first time, and compound 1 is a new triterpenoid compound, which exhibits significantly higher antitumor activity against human hepatoma cell line Bel 7402 in vitro than the blank control.
Natural product research 10/2012; 27(14). DOI:10.1080/14786419.2012.733391 · 1.23 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Qian-wang-hong-bai-san (QW), a Chinese herbal formula, is traditionally used as a skin whitening agent in China. Aim of study: In our previous screening assays, QW was identified as an effective tyrosinase inhibitor. In this study, we aim to investigate the underlying mechanism of the anti-melanogenic effect of QW in B16 cells.
Cytotoxicity of QW in B16 cell line was examined by MTT assay. Cellular tyrosinase activity was determined based on the melanin content measured at 475 nm with a microplate spectrophotometer. Protein expression was analyzed by Western blotting and quantified by Quantity One.
QW dose-dependently inhibited tyrosinase activity and decreased melanin content at 48 h without significant cytotoxicity in B16 cells. Western blot analysis showed that QW treatment down-regulated the expression levels of phospho-p38, phospho-CREB, MITF, tyrosinase, TRP-1 and TRP-2 in a dose-dependent manner. At the same time, QW treatment for 48 h inhibited IBMX-induced elevation of cellular melanin content and tyrosinase activity. However, the attenuation of IBMX-mediated up-regulations of phospho-CREB and phospho-PKA was readily observed with 60 min of QW treatment.
The anti-melanogenic activity of QW in B16 melanoma cells can be attributed, at least in part, to the inhibition of the p38 MAPK and PKA signaling pathways. These findings shed new light on the molecular mechanisms of the skin-whitening property of QW.
Journal of Ethnopharmacology 06/2012; 141(2):622-628. DOI:10.1016/j.jep.2011.08.043 · 3.00 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Norisoboldine is an alkaloid and has been identified as the major active component in Linderae Radix. A novel method for quantitative analysis of norisoboldine in Linderae Radix using ultra performance liquid chromatography (UPLC) with UV detection was developed for quality control. A similar and conventional HPLC protocol was simultaneously developed for comparison purposes. Chromatographic separation of norisoboldine was performed on an Acquity UPLC BEH C18 column (50 mm × 2.1 mm ID, 1.7 µm) for UPLC and on a Waters X-Bridge C18 column (250 mm × 4.6 mm ID, 5.0 µm) for HPLC. In the UPLC method, the average recovery was 97.3% (n = 9, RSD 1.1%); the limit of detection (LOD) and limit of quantification (LOQ) were 0.151 and 0.302 ng, respectively. In the HPLC method, the average recovery was 98.9% (n = 9, RSD 1.7%); the LOD and LOQ were 1.51 and 3.02 ng, respectively. The UPLC method provided a shorter analysis time, a better sensitivity in detection and quantitation, and less solvent was used in comparing to the HPLC method. The amount of norisoboldine detected in each Linderae Radix sample from different sources were in good agreement in both HPLC and UPLC methods.
Journal of Liquid Chromatography & Related Technologies 04/2012; 35(6):788-797. DOI:10.1080/10826076.2011.608236 · 0.64 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: 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.
[Show abstract][Hide abstract] ABSTRACT: To investigate the chemical constituents of Psidium guajava L, the EtOH/H(2)O extract of the fresh leaves was subjected to various chromatography. One diphenylmethane, one benzophenone, and eight flavonoids were isolated and elucidated as 2,6-dihydroxy-3-formaldehyde-5-methyl-4-O-(6″-O-galloyl-β-D-glucopyranosyl)-diphenylmethane (1), 2,6-dihydroxy-3,5-dimethyl-4-O-(6″-O-galloyl-β-D-glucopyranosyl)-benzophenone (2), kaempferol (3), quercetin (4), quercitrin (5), isoquercitrin (6), guaijaverin (7), avicularin (8), hyperoside (9), reynoutrin (10) by spectroscopic methods, including 1D and 2D NMR and HR-ESI-MS spectrometry as well as by comparison with published data. Compounds 5 and 10 are obtained from P. guajava for the first time, and compound 1 is a new diphenylmethane compound.
Natural product research 11/2011; 26(21):1971-5. DOI:10.1080/14786419.2011.633081 · 1.23 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: One new bibenzyl, 7, and one new diarylheptanone, diobulbinone A (18), together with sixteen known compounds, 1-6 and 8-17, have been isolated form the rhizomes of Dioscorea bulbifera. Their structures were elucidated by NMR and MS analyses. Compound 7 showed high antioxidant capacity in FRAP assay and DPPH radical-scavenging activity.