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Volumn 17, Issue 4, 2007, Pages 1112-1116

Human ACAT inhibitory effects of shikonin derivatives from Lithospermum erythrorhizon

Author keywords

ACAT inhibitor; Shikonin derivatives

Indexed keywords

(3 HYDROXYISOVALERYL)SHIKONIN; 1,4 NAPHTHOQUINONE DERIVATIVE; ACETYLSHIKONIN; CHLOROFORM; CHOLESTEROL ACYLTRANSFERASE; CHOLESTEROL ACYLTRANSFERASE INHIBITOR; ISOBUTYRYLSHIKONIN; LITHOSPERMUM EXTRACT; SHIKONIN; SHIKONIN DERIVATIVE; UNCLASSIFIED DRUG;

EID: 33846568869     PISSN: 0960894X     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.bmcl.2006.11.024     Document Type: Article
Times cited : (42)

References (24)
  • 11
    • 33846649325 scopus 로고    scopus 로고
    • note
    • 2O (9:1) to yield 2 (30 mg). Fraction 4 was purified by chromatography on silica gel (230-400 mesh, Merck, Ø50 × 150 mm), eluted with n-hexane/EtOAc (5:1) to yield 3 (70 mg).
  • 12
    • 33846602942 scopus 로고    scopus 로고
    • note
    • 3, 300 MHz): δ 176.9, 175.4, 171.6, 168.5, 167.9, 147.4, 136.3, 133.2, 133.0, 131.3, 117.6, 111.7, 111.5, 69.7, 69.0, 46.4, 32.8, 29.2, 29.0, 25.7, 17.9.
  • 15
    • 33846567297 scopus 로고    scopus 로고
    • note
    • 15. Briefly, Flp-In AC-29 cells containing five copies of integrated FRT sequence were generated by transfection of pFRT/lacZeo into AC-29 cells. FRT site-integrated clones were selected by 100 μg/mL zeocin (Invitrogen) for 1 week. Several colonies were picked and maintained in F-12 medium supplemented with 10% fetal bovine serum (FBS) and 100 μg/mL zeocin. The presence and number of FRT-site were verified by β-galactosidase assay and Southern blot analysis, respectively. To construct stable cell lines expressing hACAT-1 or hACAT-2, Flp-In AC-29 cells were plated on 60-mm dishes and transfected with 1.8 μg of the Flp-In recombinase-encoding pOG44 vector and either 0.2 μg of pcDNA5/FRT-hACAT-1 or pcDNA5/FRT-hACAT-2. After 2 days, we proceeded to select the single colony with 500 μg/mL hygromycin (Invitrogen) for 2 weeks. Colonies were picked up, expanded, and assayed for expression of hACAT-1 or -2. Established cell lines were maintained in F-12/10% FBS containing 100 μg/mL hygromycin.
  • 16
    • 33846583836 scopus 로고    scopus 로고
    • note
    • 15 a total of 30,000 cells per well were plated on 96-well culture plates and allowed to recover overnight. Assays were done with cells at least 80% confluent. Cells were incubated in Ham's F-12 medium supplemented with 1% Eagle's vitamins and 10% heat-inactivated FBS containing 5 μg/mL NBD-cholesterol as methyl-β-cyclodextrin complex with or without ACAT inhibitor for 9 h. NBD-cholesterol was added from a 5-mg/mL stock solution in methanol, and methanol concentrations in the medium did not exceed 0.1%. Flp-In AC-29 cells incubated with NBD-cholesterol were used to determine background fluorescence attributable to free NBD-cholesterol. After incubation, the medium was removed, and the cells were washed two times with cold balanced salt solution (BSS, Invitrogen). The fluorescent intensities of 96-well culture plates were read from the top using VICTOR3 fluorescent plate reader (Perkin-Elmer) equipped with 485 nm excitation and 535 nm emission filters. After measurement, cellular protein was digested through incubation with 25 μL of 0.4 N NaOH for 2 h. The fluorescent intensity was normalized by cellular protein content and calculated by subtracting the background fluorescent intensity from the total fluorescent intensity.
  • 17
    • 33846588634 scopus 로고    scopus 로고
    • note
    • 20 100 μM of MβCD was dissolved in 10 mL F-12 medium, and 6 μM NBD-cholesterol, and 10 μg/mL BSA were added to this solution with vigorous vortex. The dispersion was sonicated at room temperature for 5 min using a Fisher model 60 sonic dismembrator at setting 5. The solution was filtered with 0.45 μm membrane filter and kept in a glass tube under argon at 4 °C for up to a week.
  • 22
    • 33846637079 scopus 로고    scopus 로고
    • Ikenoya, M.; Yoshinaka, Y.; Kobayashi, H.; Kawamine, K.; Shibuya, K.; Sato, F.; Sawanobori, K.; Watanabe, T.; Miyazaki, A. Atherosclerosis. 2006, July 1; (Epub. ahead of print).


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.