二十二碳六烯酸对体外大鼠C6胶质瘤细胞促凋亡作用

目的 探讨二十二碳六烯酸（DHA）对大鼠C6胶质瘤细胞的促凋亡作用。方法 10、25、50、75和100 μmol/L DHA处理C6细胞24、48和72 h后检测细胞活力；100 μmol/L DHA处理C6细胞24 h后，应用TUNEL方法检测凋亡细胞，流式细胞术检测凋细胞的比例；应用免疫荧光和Western blotting方法检测剪切后含半胱氨酸的天冬氨酸蛋白水解酶3（cleaved Caspase-3）在凋亡细胞中的表达情况。结果 25、50、75和100 μmol/L DHA处理C6细胞24、48和72 h后，抑制细胞活力；100 μmol/L DHA处理细胞24 h后，能够检测到TUNEL阳性的凋亡细胞; 流式细胞术检测后发现，DHA能够明显上调凋亡细胞的比例；此外，细胞经100 μmol/L DHA处理后，免疫荧光能够检测到cleaved Caspase 3阳性细胞，Western blotting能够检测到cleaved Caspase-3阳性蛋白条带。结论 DHA具有促进大鼠C6胶质瘤细胞凋亡的作用。

Pro-apoptosis effects of docosahexaenoic acid on rat C6 glioma cells in vitro

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.