脂氧素A4对Aβ_(25-35)所致N2a细胞损伤的保护作用初探

Pilot Study of Protection of 5(S),6(R)-Lipoxin A4 Methyl Ester to N2a Cell Injury Caused by Aβ_(25-35)

目的:初步探讨脂氧素A4(LXA4)对β-淀粉样蛋白25-35(Aβ_(25-35))损伤N2a细胞的保护作用及其机制。方法:用不同浓度Aβ_(25-35)处理N2a细胞,建立阿尔茨海默病(AD)细胞损伤模型,通过细胞形态学观察、CCK-8法和Hoechst 33258染色来评价细胞模型是否构建成功。细胞分为对照组、模型组(20μmol/L Aβ_(25-35))和LXA4保护组(50、100、200 nmol/L)。采用CCK-8法检测N2a细胞的活性;荧光酶标仪检测N2a细胞内活性氧(ROS)水平;ELISA法检测细胞培养上清中白介素-10(IL-10)和肿瘤坏死因子-α(TNF-α)。结果:20μmol/L Aβ_(25-35)处理N2a细胞24 h可建立细胞损伤模型。LXA4保护组的细胞存活率均较模型组显著升高。与模型组相比,200 nmol/L LXA4保护组细胞内ROS水平显著降低(P<0.01);200 nmol/L LXA4保护组IL-10水平显著高于模型组(P<0.01);200 nmol/L LXA4保护组的TNF-α水平显著低于模型组(P<0.01)。结论:LXA4对Aβ_(25-35)损伤N2a细胞具有保护作用,其机制可能是抑制ROS水平以及调节炎症因子的表达。

Objective： To explore the protective effect and mechanism of LXA4 on Aβ25-35-induced injury in N2a cells. Methods： N2a cells were treated with different concentrations of Aβ25-35, and the Alzheimer＇ s disease （AD） cell model was established by the observation of cell morphology, CCK-8 method and Hoechst 33258 staining. The experiment was divided into control group, model group （20 μmol/L Aβ25-35,） and LXA4 protective group （50, 100, 200 nmol/L）. The activities of N2a cells were detected by CCK-8. The reactive oxygen species （ROS） level of N2a cells was examined by 2＇, 7＇-dichlorofluorescin diacetate （DCFH-DA） assay. The productions of interleukin-10 （IL-10） and tumor necrosis factor-α （TNF-α were measured using enzyme-linked immunosorbant assay （ELISA） kits. Results： The N2a cells treated with 20 μmol/L Aβ25-35, for 24 h could establish the AD cell model. Compared with the model group, the cell survival rate of LXA4 protective group was increased; the ROS level of LXA4 protective group （200 nmol/L） was decreased （P〈0.01）; IL-10 of LXA4 protective group （200 nmol/L） was increased （P〈0.01）; TNF-α of LXA4 protective group （200 nmol/L） was decreased （P〈0.01）. Conclusion： LXA4 plays a protective effect on the injury of N2a cells induced by Aβ25-35, and inhibiting ROS level and regulating the expression of inflammatory factors may be the underlying mechanism.