| 摘要: |
| 目的 研究突变型人胰高血糖素样肽-1(mutated human glucagon-like peptide-1，mGLP-1)对谷氨酸诱导的人神经母细胞瘤细胞SH-SY5Y细胞损伤的影响。方法 环磷酸腺苷(cAMP)试剂盒检测细胞内cAMP含量，比较mGLP-1和天然人胰高血糖素样肽-1 (natural human glucagon-like peptide-1，nGLP-1)与GLP-1受体结合的能力；以谷氨酸诱导SH-SY5Y细胞损伤，同时用mGLP-1处理SH-SY5Y细胞，采用MTT法检测细胞存活率；乳酸脱氢酶(LDH)试剂盒检测细胞LDH的释放量；DAPI染色法观察细胞凋亡形态学特征；钙流法检测胞内钙离子浓度变化；通过检测丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性、总谷胱甘肽(total GS)含量的变化判断细胞氧化损伤程度。结果 mGLP-1与nGLP-1类似，都能提高细胞内cAMP水平；mGLP-1能缓解谷氨酸诱导的细胞损伤，增加细胞存活率，降低LDH释放量；mGLP-1对谷氨酸导致的细胞内钙离子变化没有修复作用；mGLP-1能抑制氧化损伤指标MDA含量升高，促进抗氧化系统指标SOD活性增强和total GS含量增加。结论 mGLP-1可导致细胞内cAMP含量增加。mGLP-1对谷氨酸所致神经细胞损伤具有保护作用，这可能是mGLP-1通过对神经细胞的抗氧化损伤作用实现的，而不是改善谷氨酸导致的钙稳态失衡。mGLP-1的保护作用与nGLP-1类似，而mGLP-1在体内稳定性更强，半衰期更长；提示mGLP-1可能对相关神经退行性疾病的治疗具有更佳的潜在价值。 |
| 关键词: 谷氨酸中毒 突变型人胰高血糖素样肽-1 胞内钙离子 氧化损伤 神经母细胞瘤细胞 |
| DOI： |
| 分类号: |
| 基金项目:国家自然科学基金资助项目(30870790) |
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| Mutated Recombinant Human Glucagon-like Peptide-1 Protects SH-SY5Y Cells from Apoptosis Induced by Glutamate |
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华东师范大学，a.脑功能基因组学教育部重点实验室，上海市脑功能基因组学重点实验室；b.生命科学学院，上海 2000621,2
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1.East China Normal University, a.Key Laboratory of Brain Functional Genomics, Ministry of Education, Shanghai Key Laboratory of Brain Functional Genomics;2.b.School of Life Science, Shanghai 200062, China
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| Abstract: |
| OBJECTIVE To investigate the effects of mutated recombinant human glucagon-like peptide-1(mGLP-1) on glutamate-induced SH-SY5Y cell apoptosis. METHODS A cyclic adenosine monophosphate (cAMP) kit was selected for assaying intracellular cAMP content. MTT assay was applied to detect the viability of SH-SY5Y cells exposed to glutamate or /and mGLP-1. Lactate dehydrogenase (LDH) release was measured to detect glutamate-induced cytotoxicity. Apoptosis morphological identification was evaluated with 4', 6-diamidino-2-phenylindole (DAPI) nuclear staining. The cytosolic calcium concentration was tested by calcium influx assay. The oxidative damage was estimated by measuring the malondialdehyde (MDA) content, superoxide dismutase (SOD) activity and total glutathione (total GS) level. RESULTS mGLP-1 could induce the increase of intracellular cAMP content. Glutamate cytotoxicity presented that low cell viability, high LDH release and increased cell apoptosis. After treatment with mGLP-1, glutamate-induced toxicity would be partially relieved. It was evident that mGLP-1 could decrease MDA content, increase SOD activity and total GS level, but could not change calcium concentration. CONCLUSION These data indicate that mGLP-1 has protective effects on glutamate-induced SH-SY5Y cell apoptosis through rescuing oxidative damage. It is suggested that mGLP-1 may have better potential therapeutic value in the treatment of neurodegenerative diseases since mGLP-1 presents a longer half-life than natural GLP-1. |
| Key words: glutamate toxicity mutated recombinant human glucagon-like peptide-1 cytosolic calcium oxidative damage SH-SY5Y cells |
