SOD2在姜黄素减轻氧糖剥夺所致神经元损伤中的作用
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摘要
目的:探讨二型超氧化物歧化酶(Mn-SOD,SOD2)是否介导了姜黄素(Curcumin,Cur)对氧糖剥夺模型(Oxygen-Glucose Deprivation,OGD)损伤神经元的保护作用。方法:本研究采用HT22神经元细胞暴露于OGD环境中3 h模拟神经元缺血缺氧损伤,SOD2-si RNA抑制神经元SOD2蛋白表达后,通过噻唑蓝法(Methylthiazolyldiphenyl-tetrazolium bromide,MTT)检测细胞活力,比色法测量培养基乳酸脱氢酶(Lactic Dehydrogenase LDH)水平,流式细胞仪计算细胞凋亡率,Western blot测定凋亡蛋白Cleaved Caspase-3表达,并观察细胞形态和线粒体功能。结果:与正常培养的Control组相比,OGD组细胞活力显著降低,LDH释放明显增加,细胞凋亡率和Cleaved Caspase-3表达显著上升,细胞形态破坏并降低线粒体膜电位(MMP)和线粒体复合物1(Mitochondrial Complex 1 Activity)的活力(P<0.05),100 ng/ml的Cur可显著减轻OGD诱导的神经元细胞的上述损伤性改变(P<0.05)。而SOD2-si RNA显著逆转Cur对OGD诱导的神经元细胞损伤的保护作用(P<0.05),SC-si RNA则未对Cur产生的神经保护作用造成显著干扰(P>0.05)。结论:Cur可能通过上调SOD2的表达,减轻OGD对神经元细胞的损伤。
Objective: To explore whether Mn-SOD(SOD2) mediates curcumin-induced neuroprotection against oxygen-glucose deprivation(OGD) in neurons. Methods: HT22 neurons were exposed to OGD for 3 h to mimic neuronal ischemic injury, after down-regulating SOD2 expression in neurons by using SOD2-siRNA, methylthiazolyldiphenyl-tetrazolium bromide(MTT) was used to assess cell viability, reagent kit was taken to evaluate lactic dehydrogenase(LDH) in the medium, flow cytometry was taken to calculate cell apoptosis rate, western blot was used to measure cleaved caspase-3 expression, neuronal morphology and mitochondrial function were also observed. Results: Compared with the cells in the control group, OGD reduced cell viability, increased LDH release, cell apoptosis rate and cleaved caspase-3 expression, damaged cell morphology, and inhibited mitochondrial membrane potential(MMP) and complex 1 activity(P<0.05), 100 ng/mL Cur obviously reduced the OGD-induced damage mentioned above, SOD2-siRNA significantly reversed the Cur-induced neuroprotective effects(P<0.05), but the scrambled si RNA(SC-si RNA) showed little influence on the Cur-induced neuroprotective effects(P>0.05). Conclusion: Cur can reduce OGD-induced injury in neurons by up-regulating SOD2 protein.
Objective: To explore whether Mn-SOD(SOD2) mediates curcumin-induced neuroprotection against oxygen-glucose deprivation(OGD) in neurons. Methods: HT22 neurons were exposed to OGD for 3 h to mimic neuronal ischemic injury, after down-regulating SOD2 expression in neurons by using SOD2-siRNA, methylthiazolyldiphenyl-tetrazolium bromide(MTT) was used to assess cell viability, reagent kit was taken to evaluate lactic dehydrogenase(LDH) in the medium, flow cytometry was taken to calculate cell apoptosis rate, western blot was used to measure cleaved caspase-3 expression, neuronal morphology and mitochondrial function were also observed. Results: Compared with the cells in the control group, OGD reduced cell viability, increased LDH release, cell apoptosis rate and cleaved caspase-3 expression, damaged cell morphology, and inhibited mitochondrial membrane potential(MMP) and complex 1 activity(P<0.05), 100 ng/mL Cur obviously reduced the OGD-induced damage mentioned above, SOD2-siRNA significantly reversed the Cur-induced neuroprotective effects(P<0.05), but the scrambled si RNA(SC-si RNA) showed little influence on the Cur-induced neuroprotective effects(P>0.05). Conclusion: Cur can reduce OGD-induced injury in neurons by up-regulating SOD2 protein.
引文
[1]Liu X,Wu D,Wen S,et al.Mild therapeutic hypothermia protects against cerebral ischemia/reperfusion injury by inhibiting mi R-15b expression in rats[J].Brain Circ,2017,3(4):219-226
[2]Zhang C,Brandon NR,Koper K,et al.Invasion of Peripheral Immune Cells into Brain Parenchyma after Cardiac Arrest and Resuscitation[J].Aging Dis,2018,9(3):412-425
[3]Tucker LD,Lu Y,Dong Y,et al.Photobiomodulation Therapy Attenuates Hypoxic-Ischemic Injury in a Neonatal Rat Model[J].J Mol Neurosci,2018,65(4):514-526
[4]Tonge DA,de Burgh HT,Docherty R,et al.Fibronectin supports neurite outgrowth and axonal regeneration of adult brain neurons in vitro[J].Brain Res,2012,1453:8-16
[5]Lavrnja I,Parabucki A,Brkic P,et al.Repetitive hyperbaric oxygenation attenuates reactive astrogliosis and suppresses expression of inflammatory mediators in the rat model of brain injury[J].Mediators Inflamm,2015,2015:498405
[6]Schulze J,Kaiser O,Paasche G,et al.Effect of hyperbaric oxygen on BDNF-release and neuroprotection:Investigations with human mesenchymal stem cells and genetically modified NIH3T3 fibroblasts as putative cell therapeutics[J].PLo S One,2017,12(5):e0178182
[7]Kumamaru H,Lu P,Rosenzweig ES,et al.Activation of Intrinsic Growth State Enhances Host Axonal Regeneration into Neural Progenitor Cell Grafts[J].Stem Cell Reports,2018,11(4):861-868
[8]Fan C,Song Q,Wang P,et al.Curcumin Protects Against Chronic Stress-induced Dysregulation of Neuroplasticity and Depression-like Behaviors via Suppressing IL-1βPathway in Rats[J].Neuroscience2018,392:92-106
[9]Edrees NE,Galal AAA,Abdel Monaem AR,et al.Curcumin alleviates colistin-induced nephrotoxicity and neurotoxicity in rats via attenuation of oxidative stress,inflammation and apoptosis[J].Chem Biol Interact,2018,294:56-64
[10]Dai X,Zhang J,Guo G,et al.A mono-carbonyl analog of curcumin induces apoptosis in drug-resistant EGFR-mutant lung cancer through the generation of oxidative stress and mitochondrial dysfunction[J].Cancer Manag Res,2018,10:3069-3082
[11]Han Z,Chen YR,Jones CI,et al.Shear-induced reactive nitrogen species inhibit mitochondrial respiratory complex activities in cultured vascular endothelial cells[J].Am J Physiol Cell Physiol,2007,292(3):C1103-1112
[12]Liu W,Guo Q,Zhao H.Oxidative stress-elicited YY1 potentiates antioxidative response via enhancement of NRF2-driven transcriptional activity:A potential neuronal defensive mechanism against ischemia/reperfusion cerebral injury[J].Biomed Pharmacother,2018,108:698-706
[13]Zeng G,Ding W,Li Y,et al.Morroniside protects against cerebral ischemia/reperfusion injury by inhibiting neuron apoptosis and MMP2/9 expression[J].Exp Ther Med,2018,16(3):2229-2234
[14]Huang X,Ding J,Li Y,et al.Exosomes derived from PEDF modified adipose-derived mesenchymal stem cells ameliorate cerebral ischemiareperfusion injury by regulation of autophagy and apoptosis[J].Exp Cell Res,2018,371(1):269-277
[15]Levin ED.Extracellular superoxide dismutase(EC-SOD)quenches free radicals and attenuates age-related cognitive decline:opportunities for novel drug development in aging[J].Curr Alzheimer Res,2005,2(2):191-196
[16]Koltai E,Bori Z,Osvath P,et al.Master athletes have higher mi R-7,SIRT3 and SOD2 expression in skeletal muscle than age-matched sedentary controls[J].Redox Biol,2018,19:46-51
[17]Xu L,Emery JF,Ouyang YB,et al.Astrocyte targeted overexpression of Hsp72 or SOD2 reduces neuronal vulnerability to forebrain ischemia[J].Glia,2010,58(9):1042-1049
[18]Chen M,Du ZY,Zheng X,et al.Use of curcumin in diagnosis,prevention,and treatment of Alzheimer's disease[J].Neural Regen Res,2018,13(4):742-752
[19]Boland ML,Oldham S,Boland BB,et al.Nonalcoholic steatohepatitis severity is defined by a failure in compensatory antioxidant capacity in the setting of mitochondrial dysfunction[J].World J Gastroenterol,2018,24(16):1748-1765
[20]Xie CJ,Gu AP,Cai J,et al.Curcumin protects neural cells against ischemic injury in N2a cells and mouse brain with ischemic stroke[J].Brain Behav,2018,8(2):e00921
[2]Zhang C,Brandon NR,Koper K,et al.Invasion of Peripheral Immune Cells into Brain Parenchyma after Cardiac Arrest and Resuscitation[J].Aging Dis,2018,9(3):412-425
[3]Tucker LD,Lu Y,Dong Y,et al.Photobiomodulation Therapy Attenuates Hypoxic-Ischemic Injury in a Neonatal Rat Model[J].J Mol Neurosci,2018,65(4):514-526
[4]Tonge DA,de Burgh HT,Docherty R,et al.Fibronectin supports neurite outgrowth and axonal regeneration of adult brain neurons in vitro[J].Brain Res,2012,1453:8-16
[5]Lavrnja I,Parabucki A,Brkic P,et al.Repetitive hyperbaric oxygenation attenuates reactive astrogliosis and suppresses expression of inflammatory mediators in the rat model of brain injury[J].Mediators Inflamm,2015,2015:498405
[6]Schulze J,Kaiser O,Paasche G,et al.Effect of hyperbaric oxygen on BDNF-release and neuroprotection:Investigations with human mesenchymal stem cells and genetically modified NIH3T3 fibroblasts as putative cell therapeutics[J].PLo S One,2017,12(5):e0178182
[7]Kumamaru H,Lu P,Rosenzweig ES,et al.Activation of Intrinsic Growth State Enhances Host Axonal Regeneration into Neural Progenitor Cell Grafts[J].Stem Cell Reports,2018,11(4):861-868
[8]Fan C,Song Q,Wang P,et al.Curcumin Protects Against Chronic Stress-induced Dysregulation of Neuroplasticity and Depression-like Behaviors via Suppressing IL-1βPathway in Rats[J].Neuroscience2018,392:92-106
[9]Edrees NE,Galal AAA,Abdel Monaem AR,et al.Curcumin alleviates colistin-induced nephrotoxicity and neurotoxicity in rats via attenuation of oxidative stress,inflammation and apoptosis[J].Chem Biol Interact,2018,294:56-64
[10]Dai X,Zhang J,Guo G,et al.A mono-carbonyl analog of curcumin induces apoptosis in drug-resistant EGFR-mutant lung cancer through the generation of oxidative stress and mitochondrial dysfunction[J].Cancer Manag Res,2018,10:3069-3082
[11]Han Z,Chen YR,Jones CI,et al.Shear-induced reactive nitrogen species inhibit mitochondrial respiratory complex activities in cultured vascular endothelial cells[J].Am J Physiol Cell Physiol,2007,292(3):C1103-1112
[12]Liu W,Guo Q,Zhao H.Oxidative stress-elicited YY1 potentiates antioxidative response via enhancement of NRF2-driven transcriptional activity:A potential neuronal defensive mechanism against ischemia/reperfusion cerebral injury[J].Biomed Pharmacother,2018,108:698-706
[13]Zeng G,Ding W,Li Y,et al.Morroniside protects against cerebral ischemia/reperfusion injury by inhibiting neuron apoptosis and MMP2/9 expression[J].Exp Ther Med,2018,16(3):2229-2234
[14]Huang X,Ding J,Li Y,et al.Exosomes derived from PEDF modified adipose-derived mesenchymal stem cells ameliorate cerebral ischemiareperfusion injury by regulation of autophagy and apoptosis[J].Exp Cell Res,2018,371(1):269-277
[15]Levin ED.Extracellular superoxide dismutase(EC-SOD)quenches free radicals and attenuates age-related cognitive decline:opportunities for novel drug development in aging[J].Curr Alzheimer Res,2005,2(2):191-196
[16]Koltai E,Bori Z,Osvath P,et al.Master athletes have higher mi R-7,SIRT3 and SOD2 expression in skeletal muscle than age-matched sedentary controls[J].Redox Biol,2018,19:46-51
[17]Xu L,Emery JF,Ouyang YB,et al.Astrocyte targeted overexpression of Hsp72 or SOD2 reduces neuronal vulnerability to forebrain ischemia[J].Glia,2010,58(9):1042-1049
[18]Chen M,Du ZY,Zheng X,et al.Use of curcumin in diagnosis,prevention,and treatment of Alzheimer's disease[J].Neural Regen Res,2018,13(4):742-752
[19]Boland ML,Oldham S,Boland BB,et al.Nonalcoholic steatohepatitis severity is defined by a failure in compensatory antioxidant capacity in the setting of mitochondrial dysfunction[J].World J Gastroenterol,2018,24(16):1748-1765
[20]Xie CJ,Gu AP,Cai J,et al.Curcumin protects neural cells against ischemic injury in N2a cells and mouse brain with ischemic stroke[J].Brain Behav,2018,8(2):e00921
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