白藜芦醇减轻神经元氧化应激损伤的作用机制研究
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摘要
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(P<0.05),LDH释放量明显增加(P<0.05),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。
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 medium for 24 h), H2O2group(exposed to 150 μM H2O2 for 24 h), RSV+H2O2group(exposed to 25 μM RSV+150μM H2O2 for 24 h), SOD2-si RNA+RSV+H2O2group(exposed to SOD2-si RNA for 48 h and then exposed to 25 μM RSV+150 μM H2O2 for 24 h) and SC-si RNA+RSV+H2O2group(exposed to scrambled si RNA for 48 h and then exposed to 25 μM RSV+150 μM H2O2 for 24h). 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 μM H2O2 for 24 h decreased the cell viability(P<0.05), increased the LDH release and injured the cell morphology,while 25 μM RSV restored the cell viability, decreased the LDH release and ameliorated the cell morphology. However, SOD2-si RNA partially reversed the RSV-induced protective effects above, and scrambled si RNA did not abolish the cytoprotection induced by RSV. Conclusions: RSV attenuated H2O2-induced injury in HT22 cells, and SOD2 could be involved in the neuroprotection of RSV.
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 medium for 24 h), H2O2group(exposed to 150 μM H2O2 for 24 h), RSV+H2O2group(exposed to 25 μM RSV+150μM H2O2 for 24 h), SOD2-si RNA+RSV+H2O2group(exposed to SOD2-si RNA for 48 h and then exposed to 25 μM RSV+150 μM H2O2 for 24 h) and SC-si RNA+RSV+H2O2group(exposed to scrambled si RNA for 48 h and then exposed to 25 μM RSV+150 μM H2O2 for 24h). 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 μM H2O2 for 24 h decreased the cell viability(P<0.05), increased the LDH release and injured the cell morphology,while 25 μM RSV restored the cell viability, decreased the LDH release and ameliorated the cell morphology. However, SOD2-si RNA partially reversed the RSV-induced protective effects above, and scrambled si RNA did not abolish the cytoprotection induced by RSV. Conclusions: RSV attenuated H2O2-induced injury in HT22 cells, and SOD2 could be involved in the neuroprotection of RSV.
引文
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[2]Bricker-Anthony C,Hines-Beard J,Rex TS.Molecular changes and vision loss in a mouse model of closed-globe blast trauma[J].Invest Ophthalmol Vis Sci,2014,55(8):4853-4862
[3]Prasad KN,Bondy SC.Inhibition of early upstream events in prodromal Alzheimer's disease by use of targeted antioxidants[J].Curr Aging Sci,2014,7(2):77-90
[4]Wang R,Liu YY,Liu XY,et al.Resveratrol protects neurons and the myocardium by reducing oxidative stress and ameliorating mitochondria damage in a cerebral ischemia rat model[J].Cell Physiol Biochem,2014,34(3):854-864
[5]Shiozaki M,Hayakawa N,Shibata M,et al.Closer association of mitochondria with lipid droplets in hepatocytes and activation of Kupffer cells in resveratrol-treated senescence-accelerated mice[J].Histochem Cell Biol,2011,136(4):475-489
[6]Lou Z,Li P,Han K.Selenium as a Versatile Center in Fluorescence Probe for the Redox Cycle Between HCl O Oxidative Stress and H2S Repair[J].Methods Mol Biol,2015,1208:97-110
[7]Victor VM,Rovira-Llopis S,Saiz-Alarcon V,et al.Altered mitochondrial function and oxidative stress in leukocytes of anorexia nervosa patients[J].PLo S One,2014,9(9):e106463
[8]Asadpour E,Ghorbani A,Sadeghnia HR.Water-soluble compounds of lettuce inhibit DNA damage and lipid peroxidation induced by glucose/serum deprivation in N2a cells[J].Acta Pol Pharm,2014,71(3):409-413
[9]Matés JM,Pérez-Gómez C,Nú觡ez de Castro I.Antioxidant en-zymes and human diseases[J].Clin Biochem,1999,32(8):595-603
[10]Ji G,Lv K,Chen H,et al.Mi R-146a regulates SOD2 expression in H2O2stimulated PC12 cells[J].PLo S One,2013,8(7):e69351
[11]Poonyagariyagorn HK,Metzger S,Dikeman D,et al.Superoxide dismutase 3 dysregulation in a murine model of neonatal lung injury[J].Am J Respir Cell Mol Biol,2014,51(3):380-390
[12]Fridovich I,Freeman B.Antioxidant defenses in the lung[J].Annu Rev Physiol,1986,48:693-702
[13]Fukui M,Zhu BT.Mitochondrial superoxide dismutase SOD2,but not cytosolic SOD1,plays a critical role in protection against glutamate-induced oxidative stress and cell death in HT22 neuronal cells[J].Free Radic Biol Med,2010,48(6):821-830
[14]丁慧,王鹏,王颜刚.白藜芦醇联合间充质干细胞改善损伤内皮细胞的代谢记忆效应[J].中国组织工程研究,2014,18(14):2232-2237Ding Hui,Wang Peng,Wang Yan-gang.Resveratrol combined with mesenchymal stem cells improves the metabolic memory effects on endothelial cell injury[J].Chinese Journal of Tissue Engineering Research,2014,18(14):2232-2237
[15]刘宣,王海嵘,刘佳福,等.白藜芦醇对氧糖剥夺/再灌注损伤的PCI2细胞的保护作用及其作用机制[J].现代生物医学进展,2014,14(8):1423-1427Liu Xuan,Wang Hai-rong,Liu Jia-fu,et al.Protective effect of resveratrol on the injury of PC12 cells induced by OGD/R and its mechanisms[J].Progress in Modern Biomedicine,2014,14(8):1423-1427
[16]Parazzini F.Resveratrol,inositol,vitamin D and K in the prevention of cardiovascular and osteoporotic risk:a novel approach in peri-and postmenopause[J].Minerva Ginecol,2014,66(5):513-518
[17]Rossi M,Caruso F,Antonioletti R,et al.Scavenging of hydroxyl radical by resveratrol and related natural stilbenes after hydrogen peroxide attack on DNA[J].Chem Biol Interact,2013,206(2):175-185
[18]Rege SD,Kumar S,Wilson DN,et al.Resveratrol protects the brain of obese mice from oxidative damage[J].Oxid Med Cell Longev,2013,2013:419092
[19]Khan MA,Chen HC,Wan XX,et al.Regulatory effects of resveratrol on antioxidant enzymes:a mechanism of growth inhibition and apoptosis induction in cancer cells[J].Mol Cells,2013,35(3):219-225