摘要
目的以1-甲基-4-苯基吡啶离子(1-methyl-4-phenylpyridinium,MPP+)为工具药处理MN9D细胞,建立帕金森病(Pakinson’s disease,PD)的细胞模型,观察褐藻多糖硫酸酯对细胞的保护作用,并初步探讨其作用机制。方法用不同浓度(25、50、100、200μmol/L)的MPP+处理MN9D细胞36h,50μmol/L MPP+处理MN9D细胞,时间分别为12、2436、48h,建立细胞损伤模型。用0.01、0.1、1g/L褐藻多糖硫酸酯预孵育MN9D细胞1h后,加入50μmol/L MPP+共同作用36h以探讨褐藻多糖硫酸酯的保护作用。MTT法检测细胞活力、生化法测定乳酸脱氢酶(LDH)释放量,并采用二氯荧光素乙二酯(DCF-DA)染色法检测细胞内的氧化应激水平。结果随着MPP+浓度增加或作用时间延长,MN9D细胞MTT值逐渐降低。50μmol/L MPP+处理细胞36h,MTT值明显下降,LDH释放量增加。而0.1、1g/L的褐藻多糖硫酸酯预孵育1h,可明显减轻MN9D细胞的损伤,提高MTT值并降低LDH释放量,细胞形态学改变与生化实验结果一致。50μmol/L MPP+作用12h,可使MN9D细胞的氧化应激水平明显升高,而0.1、1g/L的褐藻多糖硫酸酯预处理1h,可以拮抗MPP+引起的细胞内氧化应激水平增高。结论褐藻多糖硫酸酯可以有效拮抗MPP+对MN9D细胞的损伤作用,其机制可能与褐藻多糖硫酸酯具有抗氧化活性有关。
Objective To study the neuroprotective effect and mechanism of fucoidan against MPP^＋ -induced MN9D cell damage. Methods The MN9D cell damage model was induced by treating MN9D cells with different concentrations （25, 50, 100,200μmol/L） of MPP^＋ for 36 h and with 50 μmol/L MPP^＋ for different time course（ 12, 24, 36, 48 h）. To explore the neuroprotective effects of fucoidan, the ceils were pretreated with 0.01,0.1, 1.0 g/L fucoidan for 1 h, followed by co-treatment with 50 μmol/L MPP^＋ for 36 h. The viability of MNgD cells was detected by MTT conversion and LDH leakage. Furthermore, we detected the generation of reactive oxygen species in MN9D cells to understand the underlying mechanism of the neuroprotective effects of fucoidan. Results After being treated with different concentrations of MPP^＋ for 36 h or with 50 μmol/L MPP^＋ for different time course, the cell viability of MN9D cells decreased significantly ; 50 μmol/L MPP^＋ treatment for 36 h induced a significant decrease in MTT metabolic rate and an increase in LDH leakage rate in the MN9D cells. 0.1, 1.0 g/L fucoidan pretreatment inhibited MPP^＋ -induced cell death and significantly inhibited the generation of reactive oxygen species induced by MPP^＋. Conclusion Fucoidan can protect MN9D cell from the death induced by MPP^ ＋ , and the underlying mechanism may involve the antioxidant capacity of fucoidan.
出处
《首都医科大学学报》
CAS
北大核心
2009年第4期475-480,共6页
Journal of Capital Medical University
基金
国家重点基础研究发展(973计划)计划资助项目(2006CB500700)~~
