Protective effect of KR-31378 on oxidative stress in cardiac myocytes.
In this study, we investigated whether a novel anti-ischemic KATP opener KR-31378 [(2S,3S,4R)-N"-cyano-N-(6-amino-3,4-dihydro-3-hydroxy-2-methly-2-dimethoxymethly-2H-benzopyran-4-yl)-N'-benzylguanidine] has protective effect against oxidative stress-induced death in heart-derived H9c2 cells. Cell death was induced by BSO, butionine sulfoximine, which inhibits GSH synthesis and subsequently increases reactive oxygen species (ROS) level. Cell death was quantitatively determined by measuring lactate dehydrogenase (LDH) activity and stained by Hoechst 33258. BSO-induced ROS production and mitochondrial membrane potential (MMP) were measured using 2',7'-dichlorofluorescein diacetate oxidation and rhodamine 123, respectively. Both the LDH release and the ROS elevation induced by treatment of H9c2 cells with 10 mM BSO, were significantly decreased by KR-31378. These protective effect and antioxidant effect of KR-31378 appeared to be independent on KATP channel opening. Cells exposed to BSO showed an early reduction in MMP, and this reduction in MMP was significantly reversed by treatment with KR-31378. Caspase-3 activity in BSO treated H9c2 cells was remarkably increased, and this increased caspase-3 activity was significantly reversed by KR-31378. In conclusion, our results suggest that KR-31378 can produce cardioprotective effect against oxidative stress-induced cell death through antioxidant mechanism.
Kim
Mi-Young
MY
Department of Physiology, School of Medicine, Ajou University, Suwon, Korea.
Lee
Sunkyung
S
Yi
Kyu Yang
KY
Yoo
Sung Eun
SE
Lee
Dong-Ha
DH
Lim
Hong
H
Kim
Ho Soon
HS
Lee
Soo Hwan
SH
Baik
Eun Joo
EJ
Moon
Chang-Hyun
CH
Jung
Yi-Sook
YS
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001562:Benzimidazoles / Q000737:chemistry; D019328:Buthionine Sulfoximine / Q000037:antagonists & inhibitors / Q000633:toxicity; D053148:Caspase 3; D061945:Caspase Inhibitors; D020169:Caspases / Q000378:metabolism; D002460:Cell Line; D003652:Decanoic Acids / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005452:Fluoresceins / Q000737:chemistry; D006146:Guanidines / Q000494:pharmacology*; D006880:Hydroxy Acids / Q000494:pharmacology; D007770:L-Lactate Dehydrogenase / Q000037:antagonists & inhibitors / Q000378:metabolism; D008564:Membrane Potentials / Q000187:drug effects; D008856:Microscopy, Fluorescence; D051336:Mitochondrial Membranes / Q000187:drug effects / Q000502:physiology; D032383:Myocytes, Cardiac / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects*; D026902:Potassium Channel Blockers / Q000494:pharmacology; D011714:Pyrans / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D013449:Sulfonamides / Q000494:pharmacology; D013890:Thiourea / Q000031:analogs & derivatives / Q000494:pharmacology; D013997:Time Factors
D001562:Benzimidazoles; D061945:Caspase Inhibitors; D003652:Decanoic Acids; D005452:Fluoresceins; D006146:Guanidines; D006880:Hydroxy Acids; C438686:N''-cyano-N-(6-amino-3,4-dihydro-3-hydroxy-2-methyl-2-dimethoxymethyl-2H-benzopyran-4-yl)-N'-benzylguanidine; D026902:Potassium Channel Blockers; D011714:Pyrans; D017382:Reactive Oxygen Species; D013449:Sulfonamides; C447680:hoechst 32258; D019328:Buthionine Sulfoximine; C037631:2',7'-dichlorofluorescein; C052853:5-hydroxydecanoic acid; C114623:clamikalant; D007770:L-Lactate Dehydrogenase; D053148:Caspase 3; D020169:Caspases; D013890:Thiourea