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
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摘要
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(H_2 O_2) 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 H_2 O_2(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.
Objective: To observe the effect of pine nut protein(PE) on SH-SY5 Y cells injured by hydrogen peroxide(H_2 O_2) 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 H_2 O_2(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.
引文
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[15]张慧媛,杨擎,孙佳明,等.人参提取物对过氧化氢致人神经母瘤细胞损伤的保护作用及机制研究.中国医药导报,2017,14(30):13-17
[2]于俊林,车喜泉,常纪庆.松仁的化学成分及功效.人参研究,2001,13(1):25-27
[3]赵春兰.松子贡品探微.北京林业大学学报(社会科学版),2016,15(3):7-12
[4]乔丽杰,王延让,张明.Nrf2/HO-1通路在氧化损伤保护机制中研究进展.中国职业医学,2013,40(1):82-84
[5]Tolleson C M,Fang J Y.Advances in the mechanisms of Parkinson’s disease.Discov Medicine,2013,15:61-66
[6]Cunha M P,Martín-de-Saavedra M D,Romero A,et al.Protective effect of creatine against 6-hydroxydopamine-induced cell death in human neuroblastoma SH-SY5Y cells:Involvement of intracellular signaling pathways.Neuroscience,2013,238:185-194
[7]丁宁.红松松子壳多糖的提取纯化、组成分析及抗氧化活性研究.青岛:青岛科技大学,2014
[8]张立钢,赵玉红,李莉.松仁蛋白多肽对小鼠免疫功能的影响.东北农业大学学报,2010,41(8):94-99
[9]付昊雨.红松松子壳多糖的分离纯化、结构表征及其抗氧化活性研究.长春:吉林大学,2017
[10]张曜武,丁宁.红松松子壳多糖的单糖组成及结构初步分析.化学与生物工程,2014,31(8):75-77
[11]徐鑫,毛文东,刘国艳,等.松仁营养成分及松子油理化性质和活性成分分析.营养学报,2014,36(1):99-101
[12]Zhang H,Davies K J A,Forman H J.Oxidative stress response and Nrf-2 signaling in aging.Free Radical Biology&Medicine,2015,88:314-336
[13]周燕妮,章宏,张玉媛.Nrf2在镉诱导的氧化损伤和致癌中保护作用的研究进展.蚌埠医学院学报,2017,42(8):1149-1151
[14]吴展兴,许林勇.依达拉奉对一氧化碳中毒后大鼠脑组织HO-1和NRF-2表达的影响.中国老年学杂志,2018,38(1):47-49
[15]张慧媛,杨擎,孙佳明,等.人参提取物对过氧化氢致人神经母瘤细胞损伤的保护作用及机制研究.中国医药导报,2017,14(30):13-17
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