| Project/Area Number |
11671056
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Kidney internal medicine
|
|---|
| Research Institution | Tokyo Women's Medical College |
|---|
Principal Investigator |
SHIRAGA Hiroshi Tokyo Women's Medical College Dept. of Pediatric Nephrology, School of Medicine, 医学部, 助教授 (60175396)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
AKIOKA Yuko Tokyo Women's Medical College Dept. of Pediatric Nephrology, School of Medicine Instructor, 医学部, 助手 (90212422)
YOSHIOKA Toshomasa Tokyo Women's Medical College Dept. of Pediatric Nephrology, School of Medicine Associate Professor, 医学部, 助教授 (60146438)
ITO Katumi Tokyo Women's Medical College Dept. of Pediatric Nephrology, School of Medicine Professor, 医学部, 教授 (90056771)
|
|---|
| Project Period (FY) |
1999 – 2000
|
| Project Status |
Completed (Fiscal Year 2000)
|
| Budget Amount *help |
¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 2000: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1999: ¥1,300,000 (Direct Cost: ¥1,300,000)
|
|---|
| Keywords | Carvedilol / Endothelial cell line / Toxicity / Electron paramagnetic resonance / NO donor / Fluorescence / Nitric oxide / DAF-2DA / ステロイド依存性 / グルココルチコイド抵抗性 / SSCP法 / グルココルチコイド受容体 |
|---|
| Research Abstract |
Carvedilol, an adrenoceptor blocker with antioxidant activity, was studied for its ability to interact with Nitric oxide (NO) in a cell-free condition and in an endothelial cell line (ECV304).
In a cell-free system, carvedilol attenuated NO-dependent reduction of carboxy-2-phenyl-4, 4, 5, 5-tetramethyl-imidazoline-1-oxyl-3-oxide induced by a NO donor, (±)-(E)-4-methyl-2-[(E)-hydroxyimino]-5-nitro-6-methoxy-3-hexenamide, which was determined by electron paramagnetic resonance (EPR) spectrometry. The EPR study also showed that nitrosylhemoglobin formation in rat red blood cells by the addition of NO-saturated solution was attenuated by prior incubation with 0.1 to 10 μM carvedilol. NO-induced fluorescence in 4, 5-diaminofluorescein-2 diacethyl (DAF-2DA)-loaded ECV304 cells was attenuated by carvedilol but not by labetalol. The IC50 of carvedilol for NO donor (1-hydroxy-2-oxo-3-(aminopropyl)-3-isopropyl-1-triazene or sodium nitroprusside)-induced fluorescence of DAF-2DA in ECV304 cells was 【approximately equal】1.0 x10^<-7> M, which was similar to the reported IC50 of carvedilol for the antioxidant effect.
Cell toxicity induced by a NO donor determined by the number of viable cells after 24 hour treatment with 2-2' (hydroxynitrosohydrazino) bis-ethanamine was significantly attenuated by pretreatment with 1 μM carvedilol. Both free and cell-associated carvedilol quenched NO.Because NO mediates both physiological and pathophysiological processes, NO quenching by the drug may have diverse clinical implications depending upon specific functions of local NO in tissues where carvedilol is distributed.
|
