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色钉菇粗多糖对MPP+损伤的SH-SY5Y细胞的保护作用

张雪倩, 王艳芹, 王立安, 管振龙

河北师范大学生命科学学院，石家庄 050024；河北师范大学附属中学，石家庄 050035

摘要

本文旨在研究色钉菇(Chroogomphus rutilus)粗多糖对多巴胺能神经元的保护作用。用浓度为200、400和800 μg/mL的色钉菇粗多糖对SH-SY5Y细胞进行预处理后，加入1-甲基-4-苯基吡啶离子(1-methyl-4-phenylpyridinium, MPP+)处理48 h。采用MTT法检测MPP+损伤的SH-SY5Y细胞的存活率，用Annexin V-FITC染色和流式细胞术检测细胞凋亡率。结果显示，400 μg/mL和800 μg/mL的色钉菇粗多糖预处理可显著提高MPP+损伤的SH-SY5Y细胞存活率，降低细胞凋亡率。为了排除色钉菇粗多糖通过直接结合来降低MPP+实际浓度的可能性，将多糖洗净后，再予以细胞MPP+处理，MTT结果显示800 μg/mL的色钉菇粗多糖预处理仍可提高细胞存活率。以上结果提示，色钉菇粗多糖对SH-SY5Y细胞起到有效的保护作用，其保护作用的大部分与多糖和MPP+的直接结合无明显关系，而是源于多糖对细胞的直接作用，因此色钉菇多糖有开发成防治帕金森病药物的应用前景。

关键词： 帕金森病; 多巴胺能神经元; 细胞凋亡; 保护作用; 抗氧化

分类号：R338

[Protective effects of crude polysaccharides from Chroogomphus rutilus on SH-SY5Y cells impaired by MPP(+).] [Article in Chinese]

ZHANG Xue-Qian, WANG Yan-qin, WANG Li-An, GUAN Zhen-Long

College of Life Sciences, Hebei Normal University, Shijiazhuang 050024, China； Affiliated High School of Hebei Normal University, Shijiazhuang 050035, China

Abstract

The aim of the present study was to investigate the protective effects of crude polysaccharides from Chroogomphus rutilus on dopaminergic neurons impaired by MPP(+). SH-SY5Y cells were pretreated with crude polysaccharides (200, 400 and 800 μg/mL), and then MPP(+) was added to cell medium. After 48 h of incubation, MTT method was used to detect the survival rate of SH-SY5Y cells damaged by MPP(+). Annexin V-FITC staining and flow cytometry were used to detect apoptotic rate. The results showed that pretreating SH-SY5Y cells with crude polysaccharides (400 and 800 μg/mL) increased the survival rates, and reduced the apoptotic rates of SH-SY5Y cells. To rule out the possibility that crude polysaccharides may decrease actual concentration of MPP(+) by direct binding, we washed off crude polysaccharides before MPP(+) addition. Under this experimental condition, MTT results showed the survival rates of the SH-SY5Y cells were still significantly increased by 800 μg/mL crude polysaccharides pretreatment. These results suggest a protective effect of polysaccharides on the SH-SY5Y cells. Most of this protection is contributed by direct action of polysaccharides on the cells, not by binding with MPP(+). It is indicated that the crude polysaccharides from Chroogomphus rutilus can be developed as a potential drug for Parkinson's disease prevention and treatment in the future.

Key words: Parkinson’s disease; dopaminergic neuron; apoptosis; protective effects; antioxidant

收稿日期：2012-06-19　　录用日期：2012-08-15

通讯作者：管振龙　　E-mail: zlguan@yahoo.com

引用本文：

张雪倩, 王艳芹, 王立安, 管振龙. 色钉菇粗多糖对MPP+损伤的SH-SY5Y细胞的保护作用[J]. 生理学报 2013; 65 (2): 210-216.

ZHANG Xue-Qian, WANG Yan-qin, WANG Li-An, GUAN Zhen-Long. [Protective effects of crude polysaccharides from Chroogomphus rutilus on SH-SY5Y cells impaired by MPP(+).] [Article in Chinese]. Acta Physiol Sin 2013; 65 (2): 210-216 (in Chinese with English abstract).