Abstract:

Objective  To clarify the pro-apoptotic effects of docosahexaenoicacid (DHA) on rat C6 glioma cells in vitro. Methods  After treatment of 10, 25, 50, 75 and 100 μmol/L DHA for 24, 48 and 72 hours, C6 cell viability was detected. After treatment of 100 μmol/L DHA for 24 hours, apoptotic cells were labeled by the terminal-deoxynucleotidyl transferase mediated nick end labeling（TUNEL）method, and the proportion of apoptotic cells was counted by flow cytometry. The expression of cleaved Caspase-3 in apoptotic cells was detected by immunocytochemistry and Western blotting. Results  25, 50, 75 and 100 μmol/L DHA inhibited the C6 cell viability at 24, 48 and 72 hours. The TUNEL positive cells were found after treatment of 100 μmol/L DHA for 24 hours. Flow cytometry analysis showed that the proportion of apoptotic cells was increased. Moreover, after treatment of 100 μmol/L DHA, cleaved Caspase-3 positive cells were found by immunocytochemistry. Cleaved Caspase-3 positive protein band was investigated by Western blotting. Conclusion  DHA may induce the apoptosis of C6 cells in vitro.

Key words: Docosahexaenoic acid, Glioma, Caspase3, Western blotting, Rat