Inhibition of caspases and intracellular free Ca2+ concentrations are involved in resveratrol protection against apoptosis in rat primary neuron cultures.
OBJECTIVE
To investigate the influence of resveratrol (Res), a nutritional antioxidant, on the inhibition of apoptosis in rat primary neuron cultures.


METHODS
The cultured cortical neurons of neonatal Sprague-Dawley rats were pretreated with Res (0.1, 1.0, and 10.0 micromol/L) and oxygen-glucose deprivation/reperfusion (OGD/RP) with oxygen and glucose were initiated at d 10 in vitro. Neuronal apoptosis was determined by flow cytometry, and morphological changes of neurons were observed by an electron microscope. For the mechanism studies, the intracellular free calcium concentration ([Ca2+]i) and the transcription of caspases-3 and -12 in neurons were detected by Fura 2/AM loading and real-time RT-PCR, respectively.


RESULTS
OGD/RP insult could induce an increase in the apoptotic rate of neurons (from 11.1% to 49.0%), and elicit an obvious morphological change in neurons; pretreatments with Res (0.1, 1.0, and 10.0 micromol/L, respectively) significantly reduced the elevated rate of apoptosis to 41.7%, 40.8%, and 37.4%, respectively, and ameliorated the neuronal morphological injury. Similarly, the OGD/RP insult obviously elicited the elevated levels of the [Ca2+]i and the expressions of caspases-3 and -12 mRNA in neurons. Res pretreatments markedly depressed the neuronal abnormal elevation of [Ca2+]i and the overexpression of caspases-3 and -12 mRNA in a concentration-dependent manner.


CONCLUSIONS
Res can attenuate the rat cortical neuronal apoptosis induced by OGD/RP. The mechanisms are, at least partly, due to the inhibition of the calcium overload and the overexpression of caspases-3 and -12 mRNA.
Gong
Qi-hai
QH
Department of Pharmacology, Zunyi Medical College, Zunyi 563000, China. gongqihai@yahoo.com.cn
Wang
Qian
Q
Shi
Jing-shan
JS
Huang
Xie-nan
XN
Liu
Qiong
Q
Ma
Hu
H
D000818:Animals; D000831:Animals, Newborn; D000975:Antioxidants / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D002118:Calcium / Q000378:metabolism*; D061945:Caspase Inhibitors; D020169:Caspases / Q000378:metabolism; D018929:Cell Culture Techniques; D002540:Cerebral Cortex / Q000166:cytology; D007424:Intracellular Fluid / Q000378:metabolism; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D051381:Rats; D017207:Rats, Sprague-Dawley; D000077185:Resveratrol; D013267:Stilbenes / Q000494:pharmacology*
D000975:Antioxidants; D061945:Caspase Inhibitors; D013267:Stilbenes; D020169:Caspases; D000077185:Resveratrol; D002118:Calcium