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*

doi:10.16098/j.issn.0529-1356.2018.04.001

PDF(414 KB)

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

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*

作者信息 +

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*

Author information +

文章历史 +

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。

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.

导出引用

李浩明彭敏杨清清于婷婷秦建兵韩笑金国华. 二十二碳六烯酸对体外大鼠C6胶质瘤细胞促凋亡作用[J]. 解剖学报. 2018, 49(4): 419-423 https://doi.org/10.16098/j.issn.0529-1356.2018.04.001

LI Hao-ming PENG Min YANG Qing-qing YU Ting-ting QIN Jian-bing HAN Xiao JIN Guo-hua. Pro-apoptosis effects of docosahexaenoic acid on rat C6 glioma cells in vitro[J]. Acta Anatomica Sinica. 2018, 49(4): 419-423 https://doi.org/10.16098/j.issn.0529-1356.2018.04.001

参考文献

[1］ Yanai H. Effects of N-3 polyunsaturated fatty acids on dementia ［J］. J Clin Med Res, 2017, 9(1): 1-9.
［2］ Poudyal H, Panchal SK, Diwan Ⅴ, et al. Omega-3 fatty acids and metabolic syndrome: effects and emerging mechanisms of action ［J］. Prog Lipid Res, 2011, 50(4): 372-387.
［3］ Jacobson TA. Beyond lipids: the role of omega-3 fatty acids from fish oil in the prevention of coronary heart disease ［J］. Curr Atheroscler Rep, 2007, 9(2): 145-153.
［4］ Pérez J, Ware MA, Chevalier S, et al. Dietary omega-3 fatty acids may be associated with increased neuropathic pain in nerve-injured rats ［J］. Anesth Analg, 2005, 101(2): 444-448.
［5］ Morishita M, Tanaka T, Shida T, et al. Usefulness of colon targeted DHA and EPA as novel diabetes medications that promote intrinsic GLP-1 secretion ［J］. J Control Release, 2008, 132(2): 99-104.
［6］ Contrast Ⅰ. The Impact of cholesterol, DHA, and sphingolipids on Alzheimer’s disease ［J］. Biomed Res Int, 2013(4): 814390.
［7］ Yin Y, Sui C, Meng F, et al. The omega-3 polyunsaturated fatty acid docosahexaenoic acid inhibits proliferation and progression of non-small cell lung cancer cells through the reactive oxygen species-mediated inactivation of the PI3K/Akt pathway ［J］. Lipids Health Dis, 2017, 16(1): 87-95.
［8］ Wang F, Bhat K, Doucette M, et al. Docosahexaenoic acid (DHA) sensitizes brain tumor cells to etoposide-induced apoptosis ［J］. Curr Mol Med, 2011, 11(6): 503-511.
［9］ Cahill DP, Sloan AE, Nahed BV, et al. The role of neuropathology in the management of patients with diffuse low grade glioma: a systematic review and evidence-based clinical practice guideline ［J］. J Neurooncol, 2015, 125(3): 531-549.
［10］ Nahed BV, Redjal N, Brat DJ, et al. Management of patients with recurrence of diffuse low grade glioma ［J］. J Neurooncol, 2015, 125(3): 609-630.
［11］ Ziu M, Olson JJ. Update on the evidence-based clinical practice parameter guidelines for the treatment of adults with diffuse low grade glioma: the role of initial chemotherapy ［J］. J Neurooncol, 2016, 128(3): 487-489.
［12］ Barth RF, Kaur B. Rat brain tumor models in experimental neuro-oncology: the C6, 9L, T9, RG2, F98, BT4C, RT-2 and CNS-1 gliomas ［J］. J Neurooncol, 2009, 94(3): 299-312.
［13］ Zheng X, Shen G, Yang X, et al. Most C6 cells are cancer stem cells: evidence from clonal and population analyses ［J］. Cancer Res, 2007, 67(8): 3691-3697.
［14］ Wen PSh, Gao J, Li WH, et al. Expression and significance of arrestin beta 1 in patients with glioma ［J］. Acta Anatomica Sinica, 2014, 45(6): 779-784.(in Chinese)
文普帅，高静，李文慧，等. 神经胶质瘤中β-抑制蛋白1的表达及意义［J］. 解剖学报, 2014, 45(6): 779-784.
［15］ Cohen GM. Caspases: the executioners of apoptosis ［J］. Biochem J, 1997, 326(1): 1-16.
［16］ Elmore S. Apoptosis: a review of programmed cell death ［J］. Toxicol Pathol, 2007, 35(4): 495-516.
［17］ Wu CC, Bratton SB. Regulation of the intrinsic apoptosis pathway by reactive oxygen species ［J］. Antioxid Redox Signal, 2013, 19(6): 546-558.