花色苷对Aβ_(25-35)损伤SH-SY5Y细胞保护作用的研究

PROTECTIVE EFFECT OF ANTHOCYANIN AGAINST INJURY INDUCED BY Aβ_(25-35) IN SH-SY5Y CELLS

目的观察花色苷单体矢车菊素-3-葡萄糖苷(Cy-3-G)对Aβ_(25-35)损伤SH-SY5Y细胞的保护作用,并初步探讨相关机制。方法体外培养SH-SY5Y细胞。实验分组:首先进行Aβ_(25-35)损伤浓度的筛选,将细胞分为对照组和不同浓度Aβ_(25-35)处理组(浓度为5、10、15、20、25μmol/L);在确定Aβ_(25-35)的损伤浓度基础上进行Cy-3-G干预浓度的筛选,将细胞分为对照组、Aβ_(25-35)处理组和不同浓度Cy-3-G预孵育组(浓度为10、25、50、100μmol/L);而后进行Cy-3-G干预时间的筛选,将细胞分为对照组、Aβ_(25-35)处理组和不同时间Cy-3-G预孵育组(时间为3、6、12、24h)。MTT法测定细胞活力,Hoechst染色法检测细胞凋亡率。结果与对照组比较,10μmol/L Aβ_(25-35)处理SH-SY5Y细胞时,细胞活力显著降低(P<0.05);Cy-3-G对Aβ_(25-35)处理SH-SY5Y细胞保护作用的适宜浓度和适宜作用时间分别为100μmol/L和12h;Aβ_(25-35)处理组细胞凋亡率较对照组明显升高(P<0.05),而Cy-3-G预孵育可明显抑制Aβ_(25-35)细胞凋亡。结论适宜剂量的Cy-3-G对Aβ_(25-35)损伤的SH-SY5Y细胞具有保护作用,其机制可能与抑制细胞凋亡有关。

Objective To observe the protective effect of cyanidin-3-glucoside（Cy-3-G） against injuried by Aβ_（25-35）, in SH-SY5 Y cells and investigate related mechanism. Methods SH-SY5 Y cells were cultured in vitro. First in the screening experiment of Aβ_（25-35） injury concentration, SH-SY5 Y cells were divided into control group and Aβ_（25-35）-treated groups（with Aβ_（25-35） concentrations of 5, 10, 15, 20, 25μmol/L, respectively）. Second, Cy-3-G pre- incubated concentration was screened. SH-SY5 Y cells were divided into control group, Aβ_（25-35）-treated group and Cy-3-G pre-treatment group with Cy-3-G concentrations of 10, 25, 50, 100μmol/L, respectively. Finally, the Cy-3-G intervention time was screened. SH-SY5 Y cells were divided into control group, Aβ_（25-35）-treated groups and Cy-3-G pre-treatment group（with intervention times of 3, 6, 12, 24 h, respectively）.Cell viability was measured by MTT assay. Apoptosis rate was detected by Hoechst 33342 staining method. Results Compared with control group, the cell viability in 10μmol/L Aβ_（25-35）-treated group was significantly decreased（P0.05）. For Aβ_（25-35）-treated SH-SY5 Y cells, optimal concentration and treatment time of Cy-3-G was 100μmol/L and 12 h, respectively. Compared with control group, the cell apoptosis ratio in Aβ-treated group was significantly increased, and Cy-3G pre-treatment could obviously inhibit the apoptosis caused by Aβ_（25-35）（P0.05）. Conclusion Suitable amount of Cy-3-G has protective effects against injury induced by Aβ_（25-35） in SH-SY5 Y cells, which is associated with the inhibition of apoptosis.