Sulforaphane protects primary cultures of cortical neurons against injury induced by oxygen-glucose deprivation/reoxygenation via antiapoptosis.
OBJECTIVE
To determine whether sulforaphane (SFN) protects neurons against injury caused by oxygen-glucose deprivation/reoxygenation (OGD/R) and, if so, to investigate the possible mechanisms.


METHODS
Primary cultures of neurons were prepared from the cerebral cortex of 1-day-old Sprague-Dawley rats. On days 5-6 in vitro, the neurons were exposed to OGD for 1 h, followed by reoxygenation for 24 h. Cells were treated with 0, 0.1, 0.2, 0.5, 1, 2.5, or 5 μmol/L SFN, with or without 10 μmol/L LY294002, a PI3K-specific inhibitor, during OGD/R (a total of 25 h). After 24-h reoxygenation, MTT was used to assess viability and injury was assessed by Hoechst 33258/propidium iodide (PI) staining; immunofluorescence staining and Western blot were performed to detect molecular events associated with apoptosis.


RESULTS
The MTT assay showed that 1 μmol/L SFN significantly increased viability, and Hoechst 33258/PI staining showed that the numbers of injured neurons were reduced significantly in the SFN group. Furthermore, immunofluorescence staining and Western blot showed that SFN increased Bcl-2 and decreased cleaved caspase-3 levels. Moreover, LY294002 inhibited the phosphorylated-Akt expression evoked by SFN, decreased Bcl-2 expression and increased cleaved caspase-3 expression.


CONCLUSIONS
SFN protects neurons against injury from OGD/R and this effect may be partly associated with an antiapoptosis pathway.
Department of Pathology, Chongqing Medical University, Chongqing, 400016, China.
Xuemei
X
Wu
Jing
J
Zhao
Shanshan
S
Yu
Yanlin
Y
Chen
Jingxian
J
Wu
Yong
Y
Zhao
D000818:Animals; D000831:Animals, Newborn; D016588:Anticarcinogenic Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000187:drug effects / Q000293:injuries / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D005947:Glucose / Q000172:deficiency*; D017879:Isothiocyanates; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D018696:Neuroprotective Agents / Q000494:pharmacology; D010100:Oxygen / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013454:Sulfoxides; D013861:Thiocyanates / Q000494:pharmacology*
D016588:Anticarcinogenic Agents; D017879:Isothiocyanates; D018696:Neuroprotective Agents; D013454:Sulfoxides; D013861:Thiocyanates; C016766:sulforaphane; D005947:Glucose; D010100:Oxygen