| Project/Area Number |
11671508
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Anesthesiology/Resuscitation studies
|
|---|
| Research Institution | Nagasaki University |
|---|
Principal Investigator |
TODOROKI Sachiko Medicine, Nagasaki University, Research Associate, 医学部, 助手 (50039541)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
MOROOKA Hiroaki Medicine, Nagasaki University, Associate Professor, 医学部, 助教授 (70230175)
|
|---|
| Project Period (FY) |
1999 – 2001
|
| Project Status |
Completed (Fiscal Year 2001)
|
| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2001: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2000: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1999: ¥1,900,000 (Direct Cost: ¥1,900,000)
|
|---|
| Keywords | hypoxia / protein kinase C / translocation / KCN / PC12 cell / mitochondria / 虚血 / 虚血性神経障害 / プロテインカイネースC(PKC) / PKCサブタイプ / ラット副腎髄質由来細胞 / 転写調節因子 |
|---|
| Research Abstract |
The mechanisms of neuronal degeneration following hypoxia/ischemia remain undefined, but the processes include increases in neurotransmitter release, elevation of cytosolic-free calcium concentration, and changes in signal transduction pathways. Activation of the multigene family of protein kinase C (PKC) has been associated with the release of neurotransmitter and the survival of neurons. The roles for specific PKC isozymes in regulating this response, however, are not well understood. The aim of this study was to characterize the expression of PKC isozymes and the role of PKC isozymes expression in regulating cellular damage. Therefore, to understand which PKC isozymes are involved in hypoxia/ischemia-induced neuronal degeneration, we examined PKC isozymes after hypoxia (1 % 02) or KCN exposure in rat pheochromocytoma cells (PC12 cells). A PC12 cell line was maintained in RPMI 1640 medium supplemented with 10 % (v/v) FBS, 5 % (v/v) horse serum in a humidified atmosphere containing 5 % CO2 at 37℃. Hypoxia was induced with KCN or a multigas incubator (APMW-36, ASTEC Co. Ltd, Fukuoka). The cell viability was assessed by the lactate dehydrogenase (LDH) assay. The protein levels of PKCs in the subcellular fractions were measured by immunoblot analysis. PC12 cells underwent a time-dependent decrease in cell viability after exposure to hypoxia (1 % 02) and KCN. Cell toxicity was increased significantly by KCN in glucose free RPMI 1640. Data shows that selective translocation of specific PKC isozymes occurs during hypoxia, with only calcium-dependent PKC-γ and not the other PKC isozymes increasing significantly hi the membrane and mitochondria fractions after KCN treatment of PC12 cells. Therefore, we proposed that activation/translocation of PKC-γ by hypoxia is important in the regulation of hypoxia-induced cell injury in PC 12 cells.
|
