白藜芦醇减轻神经元氧化应激损伤的作用机制研究

目的:观察白藜芦醇(RSV)对过氧化氢(H2O2)所致的海马神经元HT22细胞损伤的保护作用,并探讨超氧化物歧化酶2(Mn-SOD)在其中的作用。方法:采用体外培养HT22小鼠海马神经元细胞系,H2O2作为损伤因素模拟氧化应激损伤。将细胞分为5组,分别为正常培养组(Control)、150μM H2O2损伤组(H2O2)、25μM白藜芦醇保护组(RSV+H2O2)、SOD2-si RNA干扰组(SOD2-si RNA+RSV+H2O2)和乱序RNA组(SC-si RNA+RSV+H2O2),药物暴露24 h后,应用MTT法检测HT22细胞活力、比色法检测乳酸脱氢酶(Lactate Dehydrogenase,LDH)释放量、相差显微镜观测细胞形态。结果:与对照组相比,H2O2组的活力显著下降(P0.05),LDH释放量明显增加(P0.05),细胞形态明显破坏;25μM的RSV显著恢复了HT22细胞的活力、减少了LDH释放、改善了细胞形态,而SOD2-si RNA显著逆转了RSV引起的上述保护作用,乱序RNA(SC-si RNA)未对上述保护作用产生明显影响。结论:白藜芦醇可能通过上调SOD2减轻H2O2对HT22细胞的氧化应激损伤。

The Mechanisms of Protective Effect of Resveratrol on the H2O2-Induced Oxidative Stress Injury in HT22 Cells

Objective:To investigate the protective effect of resveratrol (RSV) on hydrogen peroxide (H2O2)-induced injury in mouse HT22 cells and the role of Mn-SOD (SOD2) in the protection.Methods:Mouse hippocampus neuron HT22 cells exposed to H2O2 for 24 h was used to mimic the oxidative injury of neuronal cells. The HT22 cells were assigned into 5 groups, including control group (cultured in drug-free mediumfor 24 h), H2O2 group (exposed to 150 uMH2O2 for 24 h), RSV+H2O2 group (exposed to 25 uMRSV+150 uMH2O2 for 24 h), SOD2-siRNA+RSV+H2O2 group (exposed to SOD2-siRNA for 48 h and then exposed to 25 uMRSV+150 uM H2O2 for 24 h) and SC-siRNA+RSV+H2O2 group (exposed to scrambled siRNA for 48 h and then exposed to 25uMRSV+150 uMH2O2 for 24 h). MTT method was used to detect the cell viability, chromatometric method was used to assess the lactate dehydrogenase (LDH) release, and phase contrast microscope was used to record the morphology of the HT22 cells.Results:Compared with the injury of the control, an exposure of 150 uMH2O2 for 24 h decreased the cell viability (P<0.05), increased the LDHrelease and injuredthe cellmorphology, while 25 uMRSVrestored the cell viability, decreased the LDHrelease and ameliorated the cell morphology. However, SOD2-siRNA partially reversed the RSV-induced protective effects above, and scrambled siRNA did not abolish the cytoprotection induced by RSV.Conclusion:RSV attenuated H2O2-induced injury in HT22 cells, and SOD2 could be involved in the neuroprotection of RSV.