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Volumn 14, Issue 13, 2004, Pages 3563-3566

Synthesis and biological evaluation of benzimidazole-4,7-diones that inhibit vascular smooth muscle cell proliferation

Author keywords

[No Author keywords available]

Indexed keywords

BENZIMIDAZOLE 4,7 DIONE; BENZIMIDAZOLE DERIVATIVE; STRESS ACTIVATED PROTEIN KINASE; UNCLASSIFIED DRUG;

EID: 2942598186     PISSN: 0960894X     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.bmcl.2004.04.051     Document Type: Article
Times cited : (30)

References (18)
  • 1
    • 0027241856 scopus 로고
    • Ross R. Nature. 362:1993;801
    • (1993) Nature , vol.362 , pp. 801
    • Ross, R.1
  • 15
    • 2942544058 scopus 로고    scopus 로고
    • note
    • 3 cells/well in 200 μL of DMEM containing 10% (v/v) fetal bovine serum in 96-well flat-bottom plates (Costar, Corning, NY, USA). After 24 h incubation, the complete medium was replaced with DMEM containing 0.2% FBS, and incubated for an additional 72 h. And then the cells were treated with test compounds in 100 μL of DMEM containing PDGF (5 ng/mL) and 5% (v/v) fetal bovine serum for 48 h. Proliferation of the cells was determined using a colorimetric assay kit based on the uptake of WST by viable cells (Premix WST-1 cell proliferation assay system, Takara Bio Inc, Otsu, Japan). The assay kit is dependent on the reduction of tetrazolium salt WST-1, which results in formation of a dark red formazan product, by various mitochondrial dehydrogenase of viable cells
  • 18
    • 2942571016 scopus 로고    scopus 로고
    • note
    • Immunoblot assay of SMC lysate: SAPK/JNK, phospho-SAPK/JNK antibodies were purchased from Cell Signaling Technology Inc. (Beverly, MA, USA). The RAoSMCs were cultured for 72 h in DMEM containing 0.2% FBS and treated with the compound 2c (1 μM). The cells were pooled and homogenized in RIPA buffer (150 mM NaCl, 50 mM Tris, pH 7.6), 1% Triton X-100, 0.1% SDS, 0.5% sodium deoxycholate, 1 mM PMSF, 1 μg/mL aprotinin, 1 μg/mL leupeptin, and 1 μg/mL pepstatin at 4°C. After incubating for 30 min on ice, insoluble materials were removed by centrifugation at 14, 000 rpm for 15 min and the protein lysate concentrations were measured by Bradford assay. The same amounts and proportions of proteins from whole cell lysates or precipitated immune complexes were resolved on SDS-PAGE and blotted onto nitrocellulose membranes. The membrane was incubated with primary antibodies overnight at 4°C, followed by HRP-conjugated secondary antibodies for 50 min at room temperature, and detected by enhance chemiluminescence (ECL) reagent


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.