Excitotoxicity effects of glutamate on human neuroblastoma SH-SY5Y cells via oxidative damage
Excitotoxicity effects of glutamate on human neuroblastoma SH-SY5Y cells via oxidative damage
复制标题
谷氨酸通过氧化损伤对人神经母细胞瘤SH-SY5Y细胞的兴奋毒性作用
DOI：
10.1007/s12264-010-0813-7
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发表时间：
2010-02-01
影响因子：
5.6
通讯作者：
Mei, Bing
中科院分区：
文献类型：
--
作者：
Sun, Zhong-Wei;Zhang, Lan;Mei, Bing
目的探讨谷氨酸对人神经母细胞瘤SH-SY 5 Y细胞兴奋性毒性作用的机制。方法采用MTT法检测SH-SY 5 Y细胞的存活率。其他损伤的分布通过乳酸脱氢酶（LDH）释放和4 '，6-二脒基-2-苯基吲哚（DAPI）染色来检测。用钙内流法测定细胞内钙离子浓度。通过细胞内谷胱甘肽含量测定、超氧化物歧化酶（SOD）含量测定和细胞外丙二醛（MDA）含量测定分析谷氨酸诱导的氧化应激。结果谷氨酸对SH-SY 5 Y细胞有损伤作用，表现为细胞活力下降，LDH释放增加，细胞形态结构改变。谷氨酸处理后20 min内细胞内钙离子浓度无明显变化，而24 h内细胞内钙离子浓度明显升高，NMDA受体拮抗剂MK 801和代谢型谷氨酸受体拮抗剂LY 341495均不能抑制这种变化。另一方面，在SH-SY 5 Y细胞中观察到谷氨酸处理的氧化损伤，包括谷胱甘肽含量和SOD活性的降低，以及MDA水平的升高，所有这些都可以被抗氧化剂丹参酮IIA（Tan IIA，中国植物丹参的主要活性成分）减轻。结论谷氨酸对人神经母细胞瘤SH-SY 5 Y细胞的毒性作用可能是通过氧化损伤而不是通过NMDA受体介导的钙稳态破坏。
Objective To investigate the mechanisms of excitotoxic effects of glutamate on human neuroblastoma SH-SY5Y cells. Methods SH-SY5Y cell viability was measured by MTT assay. Other damaged profile was detected by lactate dehydrogenase (LDH) release and by 4', 6-diamidino-2-phenylindole (DAPI) staining. The cytosolic calcium concentration was tested by calcium influx assay. The glutamate-induced oxidative stress was analyzed by cytosolic glutathione assay, superoxide dismutase (SOD) assay and extracellular malondialdehyde (MDA) assay. Results Glutamate treatment caused damage in SH-SY5Y cells, including the decrease of cell viability, the increase of LDH release and the alterations of morphological structures. Furthermore, the concentration of cytoplasmic calcium in SH-SY5Y cells was not changed within 20 min following glutamate treatment, while cytosolic calcium concentration significantly increased within 24 h after glutamate treatment, which could not be inhibited by MK801, an antagonist of NMDA receptors, or by LY341495, an antagonist of metabotropic glutamate receptors. On the other hand, oxidative damage was observed in SH-SY5Y cells treated with glutamate, including decreases in glutathione content and SOD activity, and elevation of MDA level, all of which could be alleviated by an antioxidant Tanshinone IIA (Tan IIA, a major active ingredient from a Chinese plant Salvia Miltiorrhiza Bge). Conclusion Glutamate exerts toxicity in human neuroblastoma SH-SY5Y cells possibly through oxidative damage, not through calcium homeostasis destruction mediated by NMDA receptors.