芦丁对神经细胞氧化损伤的保护作用  

Protective Effect of Rutin Against Oxidative Injury in Neuronal Cells

作　　者：张浪[1] 赖永长[2] 汪海涛[1,2] 王日康[1] 孟茜[2] 郑文华[2] 

机构地区：[1]中山大学药学院郑文华PI组,广东广州510006 [2]中山大学中山眼科中心郑文华PI组,广东广州510060

出　　处：《中药材》2014年第4期640-644,共5页Journal of Chinese Medicinal Materials

基　　金：国家自然科学基金(30670652;30711120565;30970935);广东省科技厅计划项目(36000-4202139)

摘　　要：目的:考察芦丁(RUT)对硝普钠(sodium nitroprusside,SNP)致神经细胞损伤的保护作用,并探讨其作用的信号通路。方法:通过不同剂量SNP处理PC12细胞,建立细胞损伤模型;采用噻唑蓝(MTT)法考察RUT对细胞存活的影响;以Hoechst Stain、PI Stain观察细胞形态;以蛋白印迹(Western blot)检测细胞外调节蛋白激酶(ERK1/2)磷酸化水平;在原代皮质神经元上验证RUT的神经保护作用。结果:MTT结果显示SNP能剂量依赖性地降低PC12细胞的存活,25μg/mL的RUT能显著增加SNP(800μmol/L)处理后PC12细胞的存活率(P<0.05)。Hoechst Stain、PI Stain证实RUT能显著减少PC12细胞的死亡。Western blot结果显示RUT能显著促进ERK1/2的磷酸化,在皮质神经元上也证实了RUT的保护作用。结论:RUT对SNP诱导神经细胞损伤具有显著的保护作用,其神经保护作用可能是通过ERK1/2信号通路介导的。Objective： To observe the protective effect of rutin（ RUT） on neuronal cells against sodium nitroprusside（ SNP） induced neurotoxicity.Methods： PC12 cells were treated with different concentration of SNP for 24 h and MTT assay was applied to analyze the survival rate; PC12 cells were pretreated with rutin for 1 h,and then incubated for 24 h with SNP.MTT assay,morphological observation,as well as immunofluorescence were performed to evaluate both the SNP neurotoxicity and the protective effects of RUT,Western blot was used to analyzed the level of phosphorylated extra cellular regulated protein kinases（ ERK1 /2） after treatment with RUT,the results were also testified in primary cultured neurons.Results： Results from MTT assay showed that SNP caused cell death in a concentration-dependent manner in PC12 cells.The effect of SNP was observed at 200 ~ 1 000 μmol /L and was significant at 800 μmol /L.25μmol/L rutin partly blocked the neurotoxicity of SNP by preventing PC12 cells from apoptosis.Hoechst and PI staining indicated that SNP treatment decreased the number of viable cells and induced shrinkage and aggregation of the nucleus,whereas RUT pretreatment attenuated the toxic effects of SNP,after treatment with RUT in PC12 cells,the phosphorylation of ERK1 /2 was increased and peaked at20 min.Most importantly,the protective effect of RUT on PC12 cells was confirmed on cultured neurons.Conclusion： RUT possesses protective effect against neuronal apoptosis induced by SNP and this effect may be partially related with ERK1 /2 signaling.

关 键 词：芦丁 氧化应激 神经元损伤 神经退行性疾病 

分 类 号：R285.5[医药卫生—中药学]