Antagonistic action of caffeine against LY294002-induced apoptosis in cerebellar granule neurons.
OBJECTIVE
To study the effect of caffeine on apoptosis induced by inhibition of 1-phosphatidylinositol 3-kinase in cerebellar granule neurons.


METHODS
Cerebellar granule neurons culture, agar gel electrophoresis, and stress-activated protein kinase (SAPK)/c-Jun N-terminal protein kinase (JNK) assay kit to measure SAPK/JNK activity.


RESULTS
LY294002 evoked apoptosis concentration-dependently in cerebellar granule neurons. But death resulting from LY294002 was prevented by caffeine in a concentration-dependent manner. The survival effect of caffeine was not affected by inhibitors of ryanodine-sensitive Ca2+ release, nor was it inhibited by L-type channel blockers and N-methyl-D-aspartate (NMDA) receptor blocker. In addition, RP-cAMP, H89, and KN62 were not able to inhibit the protective effect of caffeine. Phosphorylation of c-Jun was necessary for the induction of apoptosis induced by LY294002 in cerebellar granule neurons. But caffeine directly inhibited the activation of JNK and decreased phospho-c-Jun in granule neurons.


CONCLUSIONS
Caffeine inhibited the activation of JNK and decreased the phosphorylation of c-Jun to protect granule neurons from LY294002-induced apoptosis.
Department of Pharmacology, Guangzhou Medical College of PLA, Guangzhou 510315, China.
W Y
WY
Wang
M T
MT
Li
R B
RB
Pi
P X
PX
Qiu
X W
XW
Su
S Z
SZ
Lin
G M
GM
Yan
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002110:Caffeine / Q000494:pharmacology*; D002478:Cells, Cultured; D002525:Cerebellar Cortex / Q000166:cytology*; D002867:Chromones / Q000037:antagonists & inhibitors*; D053938:DNA Fragmentation; D004305:Dose-Response Relationship, Drug; D005260:Female; D008297:Male; D009025:Morpholines / Q000037:antagonists & inhibitors*; D009474:Neurons / Q000166:cytology; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D000081082:Phosphoinositide-3 Kinase Inhibitors; D051381:Rats; D017207:Rats, Sprague-Dawley
D002867:Chromones; D009025:Morpholines; D010726:Phosphodiesterase Inhibitors; D000081082:Phosphoinositide-3 Kinase Inhibitors; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; D002110:Caffeine