中国现代中药2024,Vol.26Issue(5) :807-814.DOI:10.13313/j.issn.1673-4890.20240220001

3,6?-二芥子酰基蔗糖神经保护作用研究

Studies on Neuroprotective Effects of 3,6'-Disinapoyl Sucrose

郑晓春 唐晓丽 刘雅楠 邢瀚文 印艳萍 方芳
中国现代中药2024,Vol.26Issue(5) :807-814.DOI:10.13313/j.issn.1673-4890.20240220001

3,6?-二芥子酰基蔗糖神经保护作用研究

Studies on Neuroprotective Effects of 3,6'-Disinapoyl Sucrose

郑晓春 1唐晓丽 1刘雅楠 1邢瀚文 1印艳萍 1方芳1
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作者信息

  • 1. 北京中医药大学 中药学院,北京 102488
  • 折叠

摘要

目的:研究远志中寡糖酯类成分 3,6ʹ-二芥子酰基蔗糖(DISS)对神经的保护作用.方法:将不同浓度的DISS与APP-SH-SY5Y细胞共同孵育 24 h,四甲基偶氮唑盐(MTT)法检测各组细胞的存活率,免疫荧光法检测细胞内活性氧(ROS)水平,生化法检测超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)活性,ELISA法检测细胞内和培养液中β-淀粉样蛋白(Aβ)含量,蛋白质免疫印迹法(Western blot)检测细胞内淀粉样前体蛋白(APP)、微管相关蛋白轻链 3-Ⅰ(LC3-Ⅰ)和LC3-Ⅱ的表达.结果:DISS可以提高APP-SH-SY5Y细胞的存活率(P<0.05),减少细胞内ROS的生成(P<0.01),提高SOD和GSH-Px活力(P<0.05),降低细胞内和培养液中的Aβ含量(P<0.01,P<0.05),明显升高细胞内LC3Ⅱ蛋白表达水平,升高LC3Ⅱ/LC3Ⅰ水平(P<0.05),对APP表达无明显影响.结论:DISS可能通过降低细胞内氧化应激,减少细胞Aβ表达及调节自噬发挥神经保护作用.

Abstract

Objective:To study the neuroprotective effects of 3,6'-disinapoyl sucrose(DISS),an oligosaccharide ester component of the traditional Chinese medicine Polygala tenuifolia Willd.Methods:Different concentrations of DISS were co-incubated with APP-SH-SY5Y cells for 24 h.The survival rate was detected by methyl thiazole tetrazolium(MTT)assay,The intracellular reactive oxygen species(ROS)levels were detected by immunofluorescence.The activities of superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)were detected by biochemical methods.The intracellular and culture medium contents of β-amyloid protein(Aβ)level were detected by enzyme-linked immunosorbent assay(ELISA).The expression of amyloid precursor protein(APP),microtubule-associated protein lightchain 3-Ⅰ(LC3-Ⅰ)and LC3-Ⅱ in cells were detected by western blotting.Results:DISS could increase the viability of APP-SH-SY5Y cells(P<0.05),decrease the production of intracellular ROS(P<0.01),and increase the activities of SOD and GSH-px in APP-SH-SY5Y cells(P<0.05).Meanwhile,DISS also decreased the Aβ content in APP-SH-SY5Y cells(P<0.01)and cell culture medium(P<0.05),significantly increased the expression level of intracellular LC3Ⅱ protein and increased the ratio of LC3Ⅱ/LC3 Ⅰ(P<0.05).However,DISS had no significant effect on APP expression.Conclusion:DISS may exert neuroprotective effects by reducing intracellular oxidative stress,decreasing cellular Aβ expression and regulating autophagy.

关键词

3,6ʹ-二芥子酰基蔗糖/APP-SH-SY5Y细胞/阿尔茨海默病/β淀粉样蛋白/氧化应激/自噬

Key words

3,6'-disinapoyl sucrose/APP-SH-SY5Y cells/Alzheimer's disease/β-amyloid protein/oxidative stress/autophagy

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基金项目

国家自然科学基金面上项目(81373995)

北京中医药大学校级科研发展基金(BUCM-2021-HXFZJJ-010)

出版年

2024
中国现代中药
中国中药协会,中国医药集团总公司,中国药材公司

中国现代中药

CSTPCD
影响因子:0.65
ISSN:1673-4890
参考文献量28
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