Wedelolactone

Manufactured by Merck Group

Sourced in United States

Wedelolactone is a natural compound extracted from various plant species. It functions as a chemical reagent used in laboratory research and analysis. Wedelolactone can be utilized in various analytical techniques and experiments within the scientific community.

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Lab products found in correlation

13 protocols using wedelolactone

Notch and IKK Inhibitor Screening in SCC13 Cells

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The following compounds were used for inhibition of gamma-secretase-NOTCH and Inhibitor-kappaB Kinase in experiments: DAPT (https://pubchem.ncbi.nlm.nih.gov/compound/dapt), catalog D5942, Sigma Aldrich, St Louis, MO); RO4929097 (https://pubchem.ncbi.nlm.nih.gov/compound/49867930#section=Top), catalog No. ADV465749148, Sigma Aldrich, St Louis, MO); Wedelolactone (Wedelolactone#section=Top">https://pubchem.ncbi.nlm.nih.gov/compound/Wedelolactone#section=Top), catalog #56639, Sigma Aldrich, St Louis, MO).
SCC13 cells were treated with two different Notch inhibitors, DAPT or R04929097 in different doses and measured by FACS analysis. 10μM Wedelolactone was used to treat SCC13 cells for 24 hours. Cells then harvested and stained with CD133 antibody for FACS analysis. Spheres or colonies were counted after 5 days drug treatment. Medium with drug were changed every three days.

Quan X.X., Hawk N.V., Chen W., Coupar J., Lee S., Petersen D.W., Meltzer P.S., Montemarano A., Braun M., Chen Z, & Van Waes C. (2018). Targeting Notch1 and IKKα enhanced NF-κB activation in CD133+ Skin Cancer Stem Cells. Molecular cancer therapeutics, 17(9), 2034-2048.

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Antioxidant and Cytotoxicity Assay Protocol

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2,2-Diphenyl-2-picrylhydrazyl (DPPH; Aldrich), FeCl₂·4H₂O (Fluka), trichloroacetic acid (Sigma), phenazine methosulfate (PMS; Sigma), nicotinamide adenine dinucleotide (NADH; Sigma), nitro blue tetrazolium (NBT; Sigma Aldrich), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT; Sigma), butylated hydroxytoluene (BHT; Aldrich), stigmasterol (Sigma), caffeic acid (Sigma), wedelolactone (Sigma), and ethylenediaminetetraacetate (EDTA; Sigma) were purchased from Sigma Chemical Co. (St. Louis, MO). Chlorogenic acid was purchased from Acros Organics (Thermo Fisher Scientific Inc.). Ferrozine, ferric chloride (FeCl₃), and potassium ferricyanide (K₃Fe (CN)₆) were purchased from Showa Co., Ltd. (Tokyo, Japan). Dulbecco's Modified Eagle's Medium (DMEM; Invitrogen), fetal bovine serum (FBS, Gibco), and penicillin-streptomycin were purchased from Gibco BRL (Life technology, Paisley, Scotland).

Chan C.F., Huang W.Y., Guo H.Y, & Wang B.R. (2014). Potent Antioxidative and UVB Protective Effect of Water Extract of Eclipta prostrata L.. The Scientific World Journal, 2014, 759039.

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Comprehensive Phytochemical Protocol Inventory

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Indigo, Aliso B acetate, gentiopicroside, peimine, perillaldehyde, wedelolactone, bergapten, picroside I, berberine, cinnamaldehyde, ephedrine, p-synephrine, 6-shogao, and 6-gingerol were purchased from Sigma-Aldrich. Phellodendrine was purchased from Abcam. Baicalein, baicalin, wogonin, wogonoside, and Ephedra sinica were kindly provided by Y.L. Leu (Chang Gung University, Taiwan). Scutellaria baicalensis (precursor of Baicalein, baicalin, wogonin, wogonoside) and Ephedra sinica were supplied and authenticated by Department of Pharmacy Services, Chang Gung Memorial Hospital at Taoyuan, Taiwan. See Supplementary Information for more details on compound preparation.

Lin Y.H., Luck H., Khan S., Schneeberger P.H., Tsai S., Clemente-Casares X., Lei H., Leu Y.L., Chan Y.T., Chen H.Y., Yang S.H., Coburn B., Winer S, & Winer D.A. (2019). Aryl hydrocarbon receptor agonist indigo protects against obesity-related insulin resistance through modulation of intestinal and metabolic tissue immunity. International Journal of Obesity (2005), 43(12), 2407-2421.

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Isolating and Culturing Rat Retinal Müller Cells

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Primary cultures of rat retinal Müller cells were prepared as previously described with modifications.^{29 (link)} The dissociated retinal tissue of rats at postnatal days 5 to 7 was seeded in culture dishes that contained Dulbecco's modified Eagle's medium supplemented with 5 mM D-glucose (normal glucose as a control). The cultures were forcibly pipetted every 3 days to purify the Müller cell population until the percentage of cultured Müller cells, which was identified by the GS expression, exceeded 98%.^{34 (link)} The cells were maintained in Dulbecco's modified Eagle's medium with 25 mM D-glucose, as a HG treatment, for 48 h to mimic diabetes in vitro. IL-17A (25 ng ml⁻¹) and/or the IKK inhibitor Wedelolactone (Wedel; 10 μM; Sigma-Aldrich, St Louis, MO, USA) were applied to the primary Müller cell cultures for 24 h in the HG condition. Ad-Act1-shRNA at a multiplicity of infection of 20 infected primary Müller cells in the HG medium, and Ad-GFP was used as a control. After infection for 24 h, the adenoviruses were removed, and the cells were cultured for an additional 24 h before harvest.

Qiu A.W., Bian Z., Mao P.A, & Liu Q.H. (2016). IL-17A exacerbates diabetic retinopathy by impairing Müller cell function via Act1 signaling. Experimental & Molecular Medicine, 48(12), e280-.

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Peptide-Mediated Microbeads Formulation

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RGD peptide, wedelolactone, U0126 and A6730 were purchased from Sigma-Aldrich (ON, Canada). The microbeads were purchased from Bangs Laboratory (ON, Canada).

Khakshour S., Labrecque M.P., Esmaeilsabzali H., Lee F.J., Cox M.E., Park E.J, & Beischlag T.V. (2017). Retinoblastoma protein (Rb) links hypoxia to altered mechanical properties in cancer cells as measured by an optical tweezer. Scientific Reports, 7, 7833.

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Fluorometric Proteasome Activity Assay with Copper Sulfate

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Chemicals were obtained from commercial providers: wedelolactone, dimethyl sulfoxide (DMSO), propidium iodide (PI), and copper sulfate (Sigma-Aldrich, St. Louis, MO, USA), dihydroethidium (DHE; Cayman Pharma, Ann Arbor, MI, USA), Proteasome Activity Fluorometric Assay Kit II (UPBio, Aurora, CO, USA). The pCNDA3.1-hCTR1-N-Myc plasmid was kindly provided by Dennis J. Thiele [36 (link)].

Nehybová T., Šmarda J., Daniel L., Stiborek M., Kanický V., Spasojevič I., Preisler J., Damborský J, & Beneš P. (2017). Wedelolactone Acts as Proteasome Inhibitor in Breast Cancer Cells. International Journal of Molecular Sciences, 18(4), 729.

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Synthesis and Evaluation of Cisplatin and Wedelolactone in Ovarian Cancer Cell Lines

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Cisplatin was synthesized at Discipline of Biomedical Science, School of Medicine, The University of Sydney.25 Wedelolactone was purchased from Sigma Aldrich, Sydney, Australia. All other solvents or chemicals used in this study were of analytical grade. Human ovarian cancer cell lines A2780, A2780^cisR and A2780^ZD0473 resistant were purchased from ECACC (93112519, 93112517 for A2780 and A2780^cisR respectively). A2780^ZD0473R cell line has been developed by in vitro exposure of A2780 (ECACC no: 93112519) cells to increasing concentration of the drug ZD0473 from 0.5 to 12.5 uM.

Sarwar S., Alamro A.A., Alghamdi A.A., Naeem K., Ullah S., Arif M., Yu J.Q, & Huq F. (2021). Enhanced Accumulation of Cisplatin in Ovarian Cancer Cells from Combination with Wedelolactone and Resulting Inhibition of Multiple Epigenetic Drivers. Drug Design, Development and Therapy, 15, 2211-2227.

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Mechanism of Hsp60-Mediated Apoptosis

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Luteolin, Wedelolactone, DAPI, Hoechst 33342, JC-1, Doxorubicin, poly-L-lysine solution, Meyer’s Hematoxylin solution, DPX mountant for histology and β-actin were obtained from Sigma Aldrich. Control peptide, pan-caspase inhibitor and Caspase-9 inhibitor were purchased from Calbiochem. Dharma-FECT transfection reagent and Hsp60 siRNA were purchased from Dharmacon. Fluorochrome conjugated secondary antibodies, Annexin-V Alexa Fluor 488 and ER-Tracker™ Blue-White DPX were procured from Molecular Probes-Invitrogen. ImmEdge pen (hydrophobic barrier pen), Bloxall blocking solution, DAB peroxidase substrate kit, Vectastain ABC kit were purchased from Vector Laboratories, Inc. Burlingame. Anti-human mouse Hsp60, GAPDH, and β-tubulin antibodies were purchased from Thermo-Fisher whereas, anti-human rabbit Hsp60, Cleaved PARP, Caspase-8, Caspase-9, COX-IV, PDI and Calnexin were obtained from Cell Signaling Technology, Inc. XIAP, PCNA, and HRP-conjugated secondary antibodies were purchased from Santa Cruz Biotechnology. All chemicals and antibodies were obtained from Sigma unless specified otherwise.

Arya R.K., Singh A., Yadav N.K., Cheruvu S.H., Hossain Z., Meena S., Maheshwari S., Singh A.K., Shahab U., Sharma C., Singh K., Narender T., Mitra K., Arya K.R., Singh R.K., Gayen J.R, & Datta D. (2015). Anti-breast tumor activity of Eclipta extract in-vitro and in-vivo: novel evidence of endoplasmic reticulum specific localization of Hsp60 during apoptosis. Scientific Reports, 5, 18457.

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Modulation of Calcium Signaling in Macrophages

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For calcium modulation, macrophages were treated in calcium-free DMEM (GIBCO) or DMEM with EGTA (Sigma, USA) for calcium deprivation. Cells were treated with calcium-free DMEM plus 2 mM CaCl₂ (Sigma) for calcium recovery. Then, macrophages were also treated with SKF96365, 2-aminoethyl diphenylborinate (2-APB) (Sigma), and LaCl₃ (Aladdin, China) for inhibition of store-operated calcium entry (SOCE) or with ATP to increase calcium ATPase activity. Then, LPS O55: B5 (Sigma) was added and incubated for the indicated time periods before further measurement. For chemical modulation of CaMKKβ, AMPK, SIRT1, NF-κB, and AP-1, macrophages were pretreated with STO-609, AICAR, Compound C, wedelolactone (Sigma), SR11302 (Tocris, UK), and SRT1720 and EX527 (Selleck, USA) for 1 h and then stimulated with LPS before further measurement.

Liu X., Wang N., Zhu Y., Yang Y., Chen X., Fan S., Chen Q., Zhou H, & Zheng J. (2016). Inhibition of Extracellular Calcium Influx Results in Enhanced IL-12 Production in LPS-Treated Murine Macrophages by Downregulation of the CaMKKβ-AMPK-SIRT1 Signaling Pathway. Mediators of Inflammation, 2016, 6152713.

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Inflammatory Stress-Induced Signaling Assays

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15-oxoeicosatetraenoic acid (15-oxoETE) was purchased from Cayman Biochemical (Ann Arbor, MI). Ethacrynic acid and secondary antibodies were purchased from Santa Cruz Biotechnologies (Dallas, TX). LPS derived from E.coli 0127:B8, phorbol myristoyl acetate (PMA), actin primary antibody, wedelolactone and diethyl ether were purchased from Sigma-Aldrich (St. Louis, MO). Primary HO-1 and the IKKβ Kinase assay kit were purchased from Cell Signaling (Beverly, MA). The NQO1 primary antibody was purchased from Abcam (Cambridge, MA), GCLM from Proteintech (Chicago, IL), and GAPDH from Trevigen (Gaithersburg, MD). Solvents for liquid chromatography mass spectrometry (LC-MS) including pure water and acetonitrile were purchased from Burdick and Jackson (Morristown, NJ). Glacial acetic acid, Tris(2-carboxyethyl)phosphine hydrochloride (TCEP), Optima grade methanol and chloroform were purchased from Fisher Scientific (Pittsburgh, PA). The luciferase reporter assay was purchased from Promega (Madison, WI) and the human recombinant TNFα was purchased from Life technologies (Grand Island, NY).

Snyder N.W., Golin-Bisello F., Gao Y., Blair I.A., Freeman B.A, & Wendell S.G. (2014). 15-oxoeicosatetraenoic acid is a 15-hydroxyprostaglandin dehydrogenase-derived electrophilic mediator of inflammatory signaling pathways. Chemico-biological interactions, 234, 144-153.

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