To initiate OGD, cells were moved to serum and glucose-free DMEM and maintained in an ischemic condition with 5% CO2 and 95% N2 for 3 h at 37 °C. After OGD exposure, the cells were cultured in normal medium under normoxic condition with 5% CO2 and 95% air for 24 h for reoxygenation. Cells were pretreated with different concentrations of scoparone (≥98%; Sigma-Aldrich, St. Louis, MO, USA) for 2 h, and then subjected to OGD/R. The chemical structure of scoparone was shown in Fig. 1A .
Scoparone
Manufactured by Merck Group
Sourced in United States
Scoparone is a laboratory equipment product. It is designed for use in scientific research and analysis. The core function of Scoparone is to provide a tool for researchers and scientists to conduct their experiments and investigations.
Automatically generated - may contain errors
Lab products found in correlation
Borneol, by Merck Group (1 mentions)
Valproate, by Merck Group (1 mentions)
Phenobarbital, by Merck Group (1 mentions)
Tween 80, by Merck Group (1 mentions)
Carbamazepine, by Merck Group (1 mentions)
Phenytoin, by Merck Group (1 mentions)
Rolipram, by Merck Group (1 mentions)
Scopoletin, by Merck Group (1 mentions)
Esculetin, by Fujifilm (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Hyperoside, by Merck Group (1 mentions)
Caffeic acid, by Thermo Fisher Scientific (1 mentions)
Chlorogenic acid, by Thermo Fisher Scientific (1 mentions)
Glacial acetic acid, by Junsei (1 mentions)
Ag490, by Merck Group (1 mentions)
Penicillin streptomycin, by Lonsera (1 mentions)
Palmitic acid, by Merck Group (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Anti collagen 1, by Abcam (1 mentions)
Anti gfap, by Abcam (1 mentions)
7 protocols using scoparone
1
Scoparone Protects Against OGD/R Injury
2
Anticonvulsant Interactions: Borneol and Scoparone
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All drugs—borneol, carbamazepine, phenytoin, phenobarbital and valproate—were purchased from Sigma-Aldrich (Poznań, Poland). scoparone was isolated from Artemisia umbelliformis Lam. (Asteraceae), as reported previously [19 (link)]. All the studied drugs, except for valproate (i.e., borneol, scoparone, carbamazepine, phenobarbital and phenytoin), were suspended in a 1% aqueous solution of Tween 80 (Sigma-Aldrich, Poznań, Poland), while valproate was dissolved in distilled water. The ASMs were administered intraperitoneally (ip) as follows: phenytoin—120 min, phenobarbital—60 min and carbamazepine and valproate—30 min, before the experiments, as reported earlier [39 (link),40 (link)]. borneol and scoparone were injected i.p. at increasing time periods of 15, 30, 60 and 120 min. Of note, these pretreatment times for borneol and scoparone when used separately (i.e., 15, 30, 60 and 120 min.) were based on our previously published work in order to make the results for scoparone comparable to those for other coumarins (i.e., osthole, imperatorin and xanthotoxin) tested earlier [10 (link),26 (link)]. For all the combinations with ASMs, both borneol and scoparone were administered i.p. 15 min. before the MES test and brain tissue sampling.
3
Pharmacological Modulators of Cellular Signaling
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Nω-nitro-L-arginine methyl ester hydrochloride (L-NAME), 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), L-phenylephrine (Phe), dimethyl sulfoxide, rolipram, scopoletin, and scoparone (Fig. 1 ) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Esculetin and capillarisin were purchased from Wako Pure Chemical Industries (Osaka, Japan). Udenafil was donated by Dong-A ST Company (Seoul, Korea). All other chemicals were purchased from standard suppliers.
4
Analytical Characterization of Bioactive Compounds
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Chlorogenic acid and caffeic acid were purchased from Acros Organics (Pittsburgh, PA, USA). Hyperoside and scoparone were purchased from Sigma-Aldrich (St. Louis, MO, USA). Isoquercitrin and isoChlorogenic acid A were purchased from Biopurify Phytochemicals Ltd. (Chengdu, China). The purity of the six compounds was determined to be ≥97% by HPLC analysis. HPLC-grade reagents, methanol, acetonitrile, and water were obtained from J.T.Baker (Phillipsburg, NJ, USA). Glacial acetic acid was of analytical reagent grade and was procured from Junsei (Tokyo, Japan).
5
Investigating STAT3 Signaling Regulation
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Scoparone (6,7-dimethoxycoumarin, catalog #254886) and AG490 (JAK2 inhibitor, catalog #T3434) were purchased from Sigma Aldrich (St Louis, MO, USA). PDGF (catalog #120-HD-005) was obtained from R&D Systems (McKinley Place, Minneapolis, MN, USA). Cyclin-pro-Luc (−985/+281) and constructs expressing wild-type STAT3-flag or the constitutively active form of STAT3 (STAT3C-FLAG) were kind gifts of Dr James E. Darnell (the Rockefeller University, New York, NY, USA).
6
Hepatic Steatosis Model and Scoparone Treatment
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Mouse hepatocyte AML12 and human hepatocarcinoma cell line HepG2, Huh7 were purchased from the Cell Biology Institute of Chinese Academy of Science (Shanghai, China) and cultured in DMEM with 10% FBS and 1% penicillin/streptomycin (Lonsera, Grand Island, United States) at 37°C in a humidified atmosphere containing 5% CO2.
To induce hepatic steatosis model, cells were incubated in DMEM containing 0.5 mM palmitic acid (PA) and 1% BSA (Sigma, Steinheim, Germany) for 24 h. The cells were treated with scoparone at different doses simultaneously. The cells cultured in the DMEM with 1% BSA were used as normal control. scoparone was purchased from Shanghai Winherb Pharmaceutical Technology Development Co., Ltd. (Batch No: 190623; Purity ≥98%), and initially dissolved in 50 mM dimethyl sulfoxide (DMSO). The final concentrations of DMSO were kept below 0.1% in all culture conditions.
To induce hepatic steatosis model, cells were incubated in DMEM containing 0.5 mM palmitic acid (PA) and 1% BSA (Sigma, Steinheim, Germany) for 24 h. The cells were treated with scoparone at different doses simultaneously. The cells cultured in the DMEM with 1% BSA were used as normal control. scoparone was purchased from Shanghai Winherb Pharmaceutical Technology Development Co., Ltd. (Batch No: 190623; Purity ≥98%), and initially dissolved in 50 mM dimethyl sulfoxide (DMSO). The final concentrations of DMSO were kept below 0.1% in all culture conditions.
7
Immunofluorescence Assay for Liver Fibrosis
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Scoparone, Salvianolic acid B and Hoechst 33342(nuclear counterstaining) were purchased from Sigma Chemical (Sigma-Aldrich, St. Louis, MO, USA). Anti-α-SMA, anti-collagen I, anti-desmin, and anti-GFAP were purchased from Abcam (Abcam, Cambridge, MA, USA), malondialdehyde (MDA), superoxide dismutase (SOD) were purchased from Jiancheng Bioengineering Institute (Nanjing, China), goat anti-rabbit IgG-Cy3 and goat anti-mouse IgG-Cy3 were from Com Win Biotech Co.Ltd. (Beijing, China).
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