原儿茶酸对β-淀粉样蛋白诱导神经细胞损伤的保护作用研究

NEUROPROTECTIVE EFFECT OF PROTOCATECHUIC ACID ON Β-AMYLOID-INDUCED NEURONAL INJURY IN VITRO

目的 通过β-淀粉样蛋白(β-amyloid protein,Aβ)处理SH-SY5Y细胞或原代海马神经元建立AD细胞模型,探讨原儿茶酸(protocatechuic acid,PCA)对Aβ损伤后的神经细胞的保护效应及其可能机制。方法 应用MTT法筛选Aβ25-35损伤SH-SY5Y细胞的浓度,并筛选PCA作用于Aβ25-35损伤的SH-SY5Y细胞的适宜浓度。通过DCFH-DA染色和JC-1染色探究PCA对Aβ25-35损伤的细胞内ROS水平和线粒体膜电位的影响。结果 Aβ25-35损伤SH-SY5Y细胞适宜的造模条件为10μmol/L处理24 h;800μmol/L的PCA对Aβ25-35损伤的SH-SY5Y细胞保护效果最佳(P<0.001)。Aβ25-35处理导致原代海马神经元细胞内ROS水平升高(P<0.001)而线粒体膜电位有降低趋势(P=0.287);PCA干预后,原代海马神经元的细胞内ROS水平显著降低(P<0.01),线粒体膜电位有升高趋势但无统计学差异。结论 PCA能通过减少ROS生成逆转Aβ25-35对神经细胞的毒性作用。

Objective In this study,an AD cell model was established by treating SH-SY5Y cells or primary hippocampal neurons withβ-amyloid protein(Aβ)to investigate the protective effect of protocatechuic acid(PCA)on Aβ-damaged nerve cells and its possible mechanisms.Methods MTT assay was used to screen the appropriate concentrations of Aβ25-35 and PCA treatments for SH-SY5Y cells.The effects of PCA on intracellular ROS level and mitochondrial membrane potential of Aβ25-35-damaged primary hippocampal neurons were investigated by DCFH-DA staining and JC-1 staining,respectively.Results For SH-SY5Y cells,the optimal conditions for establishing AD cell model were 10μmol/L Aβ25-3s for 24h of culture.PCA at the concentration of 800μmol/L had a best protective effect on Aβ25-35-damaged SH-SY5Y cells(P<0.001).Aβ25-35 treatment resulted in increased intracellular ROS level in primary hippocampal neurons(P<0.001).There was a decrease in mitochondrial membrane potential after Aβ25-35 treatment,but there was no statistical difference.After PCA treatment,intracellular ROS level of primary hippocampal neurons was significantly decreased(P<0.01),and the mitochondrial membrane potential increased,but there was no statistical difference.Conclusion Aβ25-3s can produce cytotoxicity by inducing ROS production.PCA can reverse the cytotoxicity of Aβ25-35 on nerve cells by reducing ROS production.