摘要
目的 观察褐藻多糖硫酸酯（Fucoidan）对人神经母细胞瘤（SH-SY5Y）细胞增殖的影响,探讨其分子机制.方法 采用噻唑蓝（MTT）法观察Fucoidan对SH-SY5Y细胞增殖、细胞糖氧剥夺损伤及糖氧剥夺/再灌注损伤的影响;采用碘化丙锭（PI）染色和流式细胞术检测Fucoidan处理后细胞周期的变化;通过Western blot检测细胞周期调控因子和凋亡调控因子的表达水平的变化.结果 低浓度Fucoidan促进细胞增殖,并呈剂量和时间依赖性,药物作用48 h后,0,6～1.0 g/L Fucoidan显著促进细胞增殖,其中0.8 g/L Fucoidan处理组比对照组细胞增殖率提高63.6％,0.8 g/L Fucoidan对SH-SY5Y细胞的糖氧剥夺损伤及糖氧剥夺/再灌注损伤均有显著保护作用,细胞增殖率比正常无损伤细胞提高13.4％;0.8 g/L Fucoidan使S期细胞比例明显增加,处理24h由10.89％增加至18.18％;提高细胞周期蛋白A（Cyclin A）、周期蛋白依赖性激酶1（CDK1）和B细胞淋巴瘤/白血病-2 （bcl-2）的表达,同时降低促凋亡蛋白bcl-2相关X蛋白（bax）的表达.结论 低浓度Fucoidan在体外能明显促进SH-SY5Y细胞的增殖,显著降低缺氧缺糖导致的细胞损伤;它可能通过促进Cyclin A、Cyclin B1、CDK1和bcl-2的表达,抑制线粒体凋亡途径中的bax基因表达,实现其对神经细胞的保护作用.
Objective To study the protective effects of the Fucoidan on human SH-SY5Y neuroblastoma cells and the molecular mechanism.Methods The effect of Fucoidan on the proliferation of SH-SY5Y cells,oxygen and glucose deprivation （OGD）,oxygen and glucose deprivation reperfusion （OGD/R） cells was tested by methyl thiazol tetrazolium （MTT） assay.Propidium iodide （PI） staining and flow cytometry were used to analyse the cell cycle distribution.The expression of several key proliferative related proteins was detected by Western blotting.Results Fucoidan at a low concentration increased the cell viability in dose-and time-dependent manners.After 48 h treatment,Fucoidan significantly promoted cell growth at a concentration ranging from 0.6 to 1.0 g/L,with 63.6％ cell growth rate compared with the control after treatment with Fucoindan of 0.8 g/L.The cell viability in both OGD and OGD/R groups treated by Fucoidan of 0.8 g/L increased significantly compared with their control groups respectively,with 13.4％ cell growth rate compared with the control.Cell cycle phase analysis revealed that Fucoindan of 0.8 g/L increased the cell percentage of S phase markedly compared with the control,from 10.89％ to 18.18％ after treatment for 24 h.Western blotting results showed that Fucoindan induced the expression of Cyclin A,cyclin dependent kinase 1 （CDK1） and B cell lymphoma/leukemia-2 （bcl-2）,as well as the decreased expression of bcl-2 associated X protein （bax）.Conclusion Fucoindan of low concentration increased the proliferation of SH-SY5Y cells significantly,which may be related to the regulation of several Cyclins,bax and bcl-2.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2015年第9期2103-2105,共3页
Chinese Journal of Experimental Surgery
基金
武汉市科技攻关计划资助项目（201260523185）
国家自然科学基金生物科学理科基地学生能力提高项目（NSFC-J1103514）
关键词
褐藻多糖硫酸酯
细胞增殖
细胞周期
糖氧剥夺
Fucoidan
Cell proliferation
Cell cycle
Oxygen and glucose deprivation
