扇贝裙边糖胺聚糖对六羟基多巴胺诱导SH-SY5Y细胞损伤的保护作用研究

发布时间：2018-02-08 20:48

本文关键词： 扇贝糖胺聚糖六羟基多巴胺神经母瘤细胞帕金森病　出处：《青岛大学》2013年硕士论文　论文类型：学位论文

【摘要】：目的：研究扇贝裙边提取物—扇贝糖胺聚糖(SS-GAG)对六羟基多巴胺(6-OHDA)诱导的多巴胺能神经元细胞损伤的保护作用,及其抗帕金森病(parkinson's disease PD)的作用机制。方法：体外培养人神经母瘤细胞株SH-SY5Y,随机分为对照组、模型组(六羟基多巴胺损伤组)和实验组(不同浓度的SS-GAG预处理细胞后,再加入六羟基多巴胺),采用MTT法测定细胞的存活率,Hochest33258荧光染色观察凋亡细胞的形态学变化,Annexin V-FITC/PI双染法检测细胞凋亡率,流式细胞术检测线粒体膜电位的改变。结果：六羟基多巴胺损伤细胞后,细胞存活率显著降低；荧光显微镜下观察细胞出现典型的凋亡形态；细胞凋亡率显著升高；线粒体膜电位显著下降。而用SS-GAG预处理24小时后,能显著提高细胞存活率；细胞凋亡程度明显减弱,凋亡率有明显下降；升高线粒体膜电位。结论：SS-GAG能够减轻六羟基多巴胺对SH-SY5Y细胞的损伤程度,且在一定浓度范围内呈剂量依赖效应。
[Abstract]:Objective: To study the protective effect of scallop skirt extract scallop glycosaminoglycan (SS-GAG) on six hydroxydopamine (6-OHDA) induced dopaminergic neuron cell injury and its mechanism of anti Parkinson's disease (parkinson's disease PD).
Methods: human neuroblastoma cell line SH-SY5Y cultured in vitro were randomly divided into control group, model group (injury group six hydroxy dopamine) and experimental group (pretreatment with various concentrations of SS-GAG cells after adding six Ohda), the survival rate of cells was determined by MTT assay, morphological changes of apoptosis was observed by Hochest33258 fluorescence Annexin V-FITC/PI staining method to detect cell apoptosis rate, mitochondrial membrane potential detection of flow cytometry.
Results: six hydroxydopamine damage cells, cell survival rate decreased significantly; under fluorescence microscope the cells appeared typical apoptotic morphology; the apoptosis rate was significantly increased; mitochondrial membrane potential decreased significantly. While pretreatment with SS-GAG after 24 hours, can significantly increase the cell survival rate; cell apoptosis significantly reduced, apoptosis rate the mitochondrial membrane potential decreased significantly; increased. Conclusion: SS-GAG can reduce six hydroxydopamine injury on SH-SY5Y cells in a dose-dependent manner, and in a certain concentration range.

【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R742.5

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