天麻素对神经母细胞瘤细胞突触相关蛋白表达的影响

Effects of gastrodin on expression of synapse-related proteins in neuroblastoma cells

目的研究天麻素(GAS)对神经母细胞瘤(SH-SY5Y)细胞突触相关蛋白表达的影响及其神经保护作用。方法采用CCK8法筛选GAS安全浓度,GAS单独或联合Aβ_(1-42)处理SH-SY5Y细胞后,采用实时荧光定量PCR及蛋白质印迹法(Western blot)分别测定突触相关蛋白突触素(SYN)、发动蛋白1(DYN1)、突触后致密蛋白95(PSD95)mRNA及蛋白表达水平。结果CCK8结果显示在0~150μmol/L GAS时细胞活力无明显变化,可作为GAS处理细胞的安全浓度;0~120μmol/L GAS处理SH-SY5Y细胞后,SYN、DYN1及PSD95 mRNA及蛋白水平均上调、并随着GAS浓度的升高而升高,差异有统计学意义(P<0.05);与对照组比较,Aβ_(1-42)组细胞SYN、DYN1、PSD95 mRNA及蛋白表达水平明显降低,差异有统计学意义(P<0.01);与Aβ组比较,Aβ+GAS组细胞SYN、DYN1、PSD95 mRNA及蛋白表达水平明显上调,差异有统计学意义(P<0.01)。结论GAS可能通过上调细胞突触相关蛋白表达来发挥神经保护作用。

Objective To study the effect of gastrodin(GAS)on the expression of synapse-related proteins and its neuroprotective effects in neuroblastoma(SH-SY5Y)cells.Methods GAS safety concentrations were screened by CCK8,and after treating SH-SY5Y cells with GAS alone or in combination with Aβ_(1-42),synapse-associated protein synaptin(SYN),initiator protein 1(DYN1),the mRNA and protein expression levels of postsynaptic compact protein 95(PSD95)were determined by quantitative real-time PCR and Western blot,respectively.Results The CCK8 results showed no significant change in cell viability at 0 to 150μmol/L GAS and served as a safe concentration in cells treated with GAS.After treating SH-SY5Y cells with 0-120μmol/L GAS,SYN,DYN1,and PSD95 mRNA and protein levels were increased and increased with increasing GAS concentration,and the difference was statistically significant(P<0.05).Compared with the control group,the SYN,DYN1,PSD95 mRNA,and protein expression levels were significantly decreased in the Aβ_(1-42) group,and the difference was statistically significant(P<0.01).Compared with group Aβ,the SYN,DYN1,PSD95 mRNA and protein expression levels were significantly upregulated in the Aβ+GAS group,and the difference was statistically significant(P<0.01).Conclusion GAS may exert a neuroprotective effect by upregulating the protein expression of cellular synapse-related.