中文摘要：

目的探讨褐藻多糖硫酸酯（FUC）对1-甲基4-苯基吡啶离子（MPP＋）损伤的MN9D细胞内氧化应激及组织蛋白酶D（CatD）单克隆凋亡相关蛋白的作用。方法采用100 mol/L MPP＋损伤MN9D细胞制备帕金森病（PD）细胞模型。观察FUC预处理后PD细胞模型的细胞存活率。采用荧光检测法测定细胞内超氧化物歧化酶（SOD）、谷胱甘肽还原酶（GSH）抑制率。采用Western blot法检测CatD、自噬相关蛋白（LC3-Ⅱ）及Bax蛋白表达。结果与MPP~＋组比较,1×10~（-6）、10~（-5）、10~（-4）mol/L FUC预处理细胞存活率均显著升高（均P〈0.01）。与对照组比较,MPP~＋组SOD、GSH抑制率显著降低（均P〈0.05）。与MPP＋组比较,FUC预保护组及SEL预保护组SOD、GSH抑制率显著升高（均P〈0.01）。FUC预保护组与SEL预保护组SOD、GSH抑制率比较差异无统计学意义。MPP~＋组CatD蛋白、LC3-Ⅱ蛋白及Bax蛋白水平表达显著高于对照组（均P〈0.001）。FUC预保护组、SEL预保护组及CatD抑制剂组CatD蛋白、LC3-Ⅱ蛋白及Bax蛋白水平显著低于MPP＋组（均P〈0.001）。FUC预保护组、SEL预保护组及CatD抑制剂组CatD蛋白、LC3-Ⅱ蛋白及Bax蛋白水平差异无统计学意义。结论 FUC对PD细胞模型有保护作用,其机制可能为保护细胞溶酶体,抑制CatDBax的表达,抗氧化应激,抑制细胞凋亡。

英文摘要：

Objective To explore the effect of fucoidan（ FUC） introcelleral oxidative stress and Cathepsin DApoptosis（ CatD） associated protein Bax of the 1-methyl-4-phenylpyridinium（ MPP~＋） intoxicated MN9D cell model.Methods PD cell model was established by MN9D cells model which damaged by 100 mol/L MPP~＋. The cell viability of PD cell model was observed after FUC pretreatment. The inhibitory rate of superoxide dismutase（ SOD）and glutathione reductase（ GSH） were measured by fluorescence detection. The expression of Bax,CatD and microtubule-associated protein light chain3（ LC3-Ⅱ） were measured by western blot. Results Compared with MPP~＋group,the cell viability were significantly increased than those after 1 × 10~（-6）,10~（-5）,10~（-4）mol/L FUC pretreatment（ all P 0. 01）. Compared with control group,the inhibitory rate of SOD and GSH in MPP＋group were significantly decreased（ all P 0. 05）. Compared with MPP＋group,the inhibitory rate of SOD and GSH in FUC pretreatment group and SEL pretreatment group were significantly higher（ all P 0. 01）. There was no significant difference in inhibitory rate of SOD and GSH between FUC pretreatment group and SEL pretreatment group. The expression of CatD,LC3-Ⅱ and Bax protein in MPP＋group were significantly increased than those in control group（ all P 0. 001）. The expression of CatD,LC3-Ⅱ and Bax protein in FUC pretreatment group,SEL pretreatment group and CatD inhibitor group were significantly decreased than those in MPP＋group（ all P 0. 001）. There was no significant difference of expression of CatD,LC3-Ⅱand Bax protein between FUC pretreatment group,SEL pretreatment group and CatD inhibitor group. Conclusions FUC has protective effect on PD cell model. The mechanism are protecting cell lysosomes,inhibiting expression of CatD-Bax,reducing oxidative stress and inhibiting apoptosis.