松子蛋白对过氧化氢致SH-SY5Y细胞氧化损伤的保护作用及其机制

Protective effect of pine nut protein on oxidative damage of SH-SY5Y cells induced by hydrogen peroxide and its mechanism

目的:观察松子蛋白(PE)对过氧化氢(H_2O_2)致SH-SY5Y细胞损伤的影响及其机制。方法:取对数生长期细胞,分为对照组,模型组,PE低、中、高剂量组(0.1、1、10μg/mL),对照组加入等体积细胞1640培养液,然后加入H_2O_2(终浓度400μmol/L)且孵育4h造成细胞氧化应激损伤模型,采用MTT比色法检测细胞活力,利用流式细胞技术检测PE对氧化损伤SH-SY5Y细胞凋亡程度的影响,Western blot检测Bcl-2、Nrf-2、HO-1蛋白表达的情况。结果:与对照组比较,模型组SH-SY5Y细胞存活率显著降低(P<0.05),凋亡率显著升高(P<0.05),SH-SY5Y中Nrf-2、Bcl-2、HO-1蛋白水平显著降低(P<0.05);与模型组比较,PE低、中、高剂量组细胞存活率均显著升高(P<0.05),凋亡率显著降低(P<0.05),SH-SY5Y中Nrf-2、Bcl-2、HO-1蛋白水平均显著升高(P<0.05)。结论:PE对H_2O_2致SH-SY5Y细胞损伤具有保护作用,其机制可能与通过激活神经细胞内Nrf-2通路抑制其氧化应激,降低凋亡发生有关。

Objective:To observe the effect of pine nut protein(PE)on SH-SY5 Y cells injured by hydrogen peroxide(H2 O2)and its mechanism.Methods:The logarithmic growth phase cells were divided into control group,model group,PE low,medium and high dose groups(0.1,1,10μg/mL).The control group was added with equal volume of 1640 medium,then H2 O2(final concentration 400μmol/L)and incubation for 4 h resulted in a model of cellular oxidative stress injury,and cell viability was detected by MTT colorimetry.The effect of PE on the apoptosis of SH-SY5 Y cells was detected by flow cytometry.The expression of Bcl-2,Nrf-2 and HO-1 protein was detected by Western blot.Results:Compared with the control group,the survival rate of SH-SY5 Y cells in the model group was significantly decreased(P<0.05),the apoptosis rate was significantly increased(P<0.05),and Nrf-2,Bcl-2,HO-1 protein levels in SH-SY5 Y were significantly reduced(P<0.05);Compared with the model group,the survival rate of PE in the low,medium and high dose groups were significantly increased(P<0.05),the apoptosis rate were significantly decreased(P<0.05),the Nrf-2,Bcl-2 and HO-1 protein levels were significantly increased(P<0.05).Compared with blank group,the apoptotic rate of model group was significantly higher(P<0.05);Compared with model group,the apoptotic rate of PE D group,PE Z group,and PE H group was significantly lower(P<0.05).Conclusion:PE has a protective effect on human neuroblastoma cell injury induced by hydrogen peroxide.Its mechanism may be related to inhibiting oxidative stress and reducing apoptosis by activating Nrf-2 pathway in neurons.