解剖学报

解剖学报 ›› 2018, Vol. 49 ›› Issue (4): 419-423.doi: 10.16098/j.issn.0529-1356.2018.04.001

• 神经生物学 •    下一篇

二十二碳六烯酸对体外大鼠C6胶质瘤细胞促凋亡作用

李浩明 彭敏 杨清清 于婷婷 秦建兵 韩笑 金国华*   

  1. 南通大学医学院人体解剖学系，神经生物学研究所，炎症微环境协同创新中心， 江苏 南通 226001
  • 收稿日期:2017-05-19 修回日期:2017-06-26 出版日期:2018-08-06 发布日期:2018-08-06
  • 通讯作者: 金国华 E-mail:jguohua@ntu.edu.cn
  • 基金资助:
    江苏省自然科学基金资助项目;国家自然科学基金资助项目;江苏高校优势学科建设工程资助项目;南通大学研究生创新计划项目

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

LI Hao-ming PENG Min YANG Qing-qing YU Ting-ting QIN Jian-bing HAN Xiao JIN Guo-hua*   

  1. Department of Anatomy, Institute of Neurobiology, Collaborative Innovation Center of Inflammatory Microenviroment, Medical School, Nantong University, Jiangsu Nantong 226001, China
  • Received:2017-05-19 Revised:2017-06-26 Online:2018-08-06 Published:2018-08-06
  • Contact: JIN Guo-hua E-mail:jguohua@ntu.edu.cn

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摘要：

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。

关键词: 二十二碳六烯酸, 胶质瘤, 含半胱氨酸的天冬氨酸蛋白水解酶3, 免疫印迹法, 大鼠

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