Deferoxamine promotes recovery of traumatic spinal cord injury by inhibiting ferroptosis
|
摘要
Ferroptosis is an iron-dependent novel cell death pathway. Deferoxamine, a ferroptosis inhibitor, has been reported to promote spinal cord injury repair. It has yet to be clarified whether ferroptosis inhibition represents the mechanism of action of Deferoxamine on spinal cord injury recovery. A rat model of Deferoxamine at thoracic 10 segment was established using a modified Allen's method. Ninety 8-week-old female Wistar rats were used. Rats in the Deferoxamine group were intraperitoneally injected with 100 mg/kg Deferoxamine 30 minutes before injury. Simultaneously, the Sham and Deferoxamine groups served as controls. Drug administration was conducted for 7 consecutive days. The results were as follows:(1) Electron microscopy revealed shrunken mitochondria in the spinal cord injury group.(2) The Basso, Beattie and Bresnahan locomotor rating score showed that recovery of the hindlimb was remarkably better in the Deferoxamine group than in the spinal cord injury group.(3) The iron concentration was lower in the Deferoxamine group than in the spinal cord injury group after injury.(4) Western blot assay revealed that, compared with the spinal cord injury group, GPX4, xCT, and glutathione expression was markedly increased in the Deferoxamine group.(5) Real-time polymerase chain reaction revealed that, compared with the Deferoxamine group, mRNA levels of ferroptosis-related genes Acyl-CoA synthetase family member 2(ACSF2) and iron-responsive element-binding protein 2(IREB2) were up-regulated in the Deferoxamine group.(6) Deferoxamine increased survival of neurons and inhibited gliosis. These findings confirm that Deferoxamine can repair spinal cord injury by inhibiting ferroptosis. Targeting ferroptosis is therefore a promising therapeutic approach for spinal cord injury.
Ferroptosis is an iron-dependent novel cell death pathway. Deferoxamine, a ferroptosis inhibitor, has been reported to promote spinal cord injury repair. It has yet to be clarified whether ferroptosis inhibition represents the mechanism of action of Deferoxamine on spinal cord injury recovery. A rat model of Deferoxamine at thoracic 10 segment was established using a modified Allen's method. Ninety 8-week-old female Wistar rats were used. Rats in the Deferoxamine group were intraperitoneally injected with 100 mg/kg Deferoxamine 30 minutes before injury. Simultaneously, the Sham and Deferoxamine groups served as controls. Drug administration was conducted for 7 consecutive days. The results were as follows:(1) Electron microscopy revealed shrunken mitochondria in the spinal cord injury group.(2) The Basso, Beattie and Bresnahan locomotor rating score showed that recovery of the hindlimb was remarkably better in the Deferoxamine group than in the spinal cord injury group.(3) The iron concentration was lower in the Deferoxamine group than in the spinal cord injury group after injury.(4) Western blot assay revealed that, compared with the spinal cord injury group, GPX4, xCT, and glutathione expression was markedly increased in the Deferoxamine group.(5) Real-time polymerase chain reaction revealed that, compared with the Deferoxamine group, mRNA levels of ferroptosis-related genes Acyl-CoA synthetase family member 2(ACSF2) and iron-responsive element-binding protein 2(IREB2) were up-regulated in the Deferoxamine group.(6) Deferoxamine increased survival of neurons and inhibited gliosis. These findings confirm that Deferoxamine can repair spinal cord injury by inhibiting ferroptosis. Targeting ferroptosis is therefore a promising therapeutic approach for spinal cord injury.
Ferroptosis is an iron-dependent novel cell death pathway. Deferoxamine, a ferroptosis inhibitor, has been reported to promote spinal cord injury repair. It has yet to be clarified whether ferroptosis inhibition represents the mechanism of action of Deferoxamine on spinal cord injury recovery. A rat model of Deferoxamine at thoracic 10 segment was established using a modified Allen's method. Ninety 8-week-old female Wistar rats were used. Rats in the Deferoxamine group were intraperitoneally injected with 100 mg/kg Deferoxamine 30 minutes before injury. Simultaneously, the Sham and Deferoxamine groups served as controls. Drug administration was conducted for 7 consecutive days. The results were as follows:(1) Electron microscopy revealed shrunken mitochondria in the spinal cord injury group.(2) The Basso, Beattie and Bresnahan locomotor rating score showed that recovery of the hindlimb was remarkably better in the Deferoxamine group than in the spinal cord injury group.(3) The iron concentration was lower in the Deferoxamine group than in the spinal cord injury group after injury.(4) Western blot assay revealed that, compared with the spinal cord injury group, GPX4, xCT, and glutathione expression was markedly increased in the Deferoxamine group.(5) Real-time polymerase chain reaction revealed that, compared with the Deferoxamine group, mRNA levels of ferroptosis-related genes Acyl-CoA synthetase family member 2(ACSF2) and iron-responsive element-binding protein 2(IREB2) were up-regulated in the Deferoxamine group.(6) Deferoxamine increased survival of neurons and inhibited gliosis. These findings confirm that Deferoxamine can repair spinal cord injury by inhibiting ferroptosis. Targeting ferroptosis is therefore a promising therapeutic approach for spinal cord injury.
引文
Angeli JPF,Shah R,Pratt DA,Conrad M(2017)Ferroptosis inhibition:mechanisms and opportunities.Trends Pharmacol Sci 38:489-498.
Basso DM,Beattie MS,Bresnahan JC(1995)A sensitive and reliable locomotor rating scale for open field testing in rats.J Neurotrauma12:1-21.
Bazinet RP,Laye S(2014)Polyunsaturated fatty acids and their metabolites in brain function and disease.Nat Rev Neurosci 15:771-785.
Cakir O,Erdem K,Oruc A,Kilinc N,Eren N(2003)Neuroprotective effect of N-acetylcysteine and hypothermia on the spinal cord ischemia-reperfusion injury.Cardiovasc Surg 11:375-379.
Chen L,Hambright WS,Na R,Ran Q(2015)Ablation of the ferroptosis inhibitor glutathione peroxidase 4 in neurons results in rapid motor neuron degeneration and paralysis.J Biol Chem 290:28097-28106.
Choi DC,Lee JY,Lim EJ,Baik HH,Oh TH,Yune TY(2012)Inhibition of ROS-induced p38MAPK and ERK activation in microglia by acupuncture relieves neuropathic pain after spinal cord injury in rats.Exp Neurol 236:268-282.
Cozza G,Rossetto M,Bosello-Travain V,Maiorino M,Roveri A,Toppo S,Zaccarin M,Zennaro L,Ursini F(2017)Glutathione peroxidase 4-catalyzed reduction of lipid hydroperoxides in membranes:the polar head of membrane phospholipids binds the enzyme and addresses the fatty acid hydroperoxide group toward the redox center.Free Radic Biol Med 112:1-11.
Dixon SJ,Lemberg KM,Lamprecht MR,Skouta R,Zaitsev EM,Gleason CE,Patel DN,Bauer AJ,Cantley AM,Yang WS,Morrison B3rd,Stockwell BR(2012)Ferroptosis:an iron-dependent form of nonapoptotic cell death.Cell 149:1060-1072.
Do Van B,Gouel F,Jonneaux A,Timmerman K,Gele P,Petrault M,Bastide M,Laloux C,Moreau C,Bordet R,Devos D,Devedjian JC(2016)Ferroptosis,a newly characterized form of cell death in Parkinson’s disease that is regulated by PKC.Neurobiol Dis 94:169-178.
Fan B,Wei Z,Yao X,Shi G,Cheng X,Zhou X,Zhou H,Ning G,Kong X,Feng S(2018)Microenvironment imbalance of spinal cord injury.Cell Transplant 27:853-866.
Fan H,Zhang K,Shan L,Kuang F,Chen K,Zhu K,Ma H,Ju G,Wang YZ(2016)Reactive astrocytes undergo M1 microglia/macrohpages-induced necroptosis in spinal cord injury.Mol Neurodegener11:14.
Genovese T,Mazzon E,Esposito E,Muia C,Di Paola R,Di Bella P,Bramanti P,Cuzzocrea S(2007)Role of endogenous glutathione in the secondary damage in experimental spinal cord injury in mice.Neurosci Lett 423:41-46.
Girotti AW(1998)Lipid hydroperoxide generation,turnover,and effector action in biological systems.J Lipid Res 39:1529-1542.
Grossman SD,Rosenberg LJ,Wrathall JR(2001a)Relationship of altered glutamate receptor subunit mRNA expression to acute cell loss after spinal cord contusion.Exp Neurol 168:283-289.
Grossman SD,Rosenberg LJ,Wrathall JR(2001b)Temporal-spatial pattern of acute neuronal and glial loss after spinal cord contusion.Exp Neurol 168:273-282.
Guiney SJ,Adlard PA,Bush AI,Finkelstein DI,Ayton S(2017)Ferroptosis and cell death mechanisms in Parkinson’s disease.Neurochem Int 104:34-48.
Hao J,Li B,Duan HQ,Zhao CX,Zhang Y,Sun C,Pan B,Liu C,Kong XH,Yao X,Feng SQ(2017)Mechanisms underlying the promotion of functional recovery by deferoxamine after spinal cord injury in rats.Neural Regen Res 12:959-968.
He M,Ding Y,Chu C,Tang J,Xiao Q,Luo ZG(2016)Autophagy induction stabilizes microtubules and promotes axon regeneration after spinal cord injury.Proc Natl Acad Sci U S A 113:11324-11329.
Huang H,Young W,Ziad A,Hooshang S,Alok S,Dafin M,Shiqing F,Lin C(2015)Beijing declaration of international association of neurorestoratology.J Neurorestoratol 3:121-122.
Imai H,Matsuoka M,Kumagai T,Sakamoto T,Koumura T(2017)Lipid peroxidation-dependent cell death regulated by GPx4 and ferroptosis.Curr Top Microbiol Immunol 403:143-170.
Koozekanani SH,Vise WM,Hashemi RM,McGhee RB(1976)Possible mechanisms for observed pathophysiological variability in experimental spinal cord injury by the method of Allen.J Neurosurg44:429-434.
Li Q,Han X,Lan X,Gao Y,Wan J,Durham F,Cheng T,Yang J,et al.(2017),Inhibition of neuronal ferroptosis protects hemorrhagic brain.JCI Insight 2:e90777.
Liu JM,Wang FC,Zhou YJ,Mu L,Hou SK,Hao LN,Zhang ZT(2016)Effect of electroacupuncture stimulation on apoptosis of nerve cells in a rat model of spinal cord contusion.Zhongguo Zuzhi Gongcheng Yanjiu 20:616-621.
Liu XZ,Xu XM,Hu R,Du C,Zhang SX,McDonald JW,Dong HX,Wu YJ,Fan GS,Jacquin MF,Hsu CY,Choi DW(1997)Neuronal and glial apoptosis after traumatic spinal cord injury.J Neurosci 17:5395-5406.
Livak KJ,Schmittgen TD(2001)Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T))Method.Methods 25:402-408.
Lu J,Ashwell KW,Waite P(2000)Advances in secondary spinal cord injury:role of apoptosis.Spine(Phila Pa 1976)25:1859-1866.
Martin-Bastida A,Ward RJ,Newbould R,Piccini P,Sharp D,Kabba C,Patel MC,Spino M,Connelly J,Tricta F,Crichton RR,Dexter DT(2017a)Brain iron chelation by deferiprone in a phase 2 randomised double-blinded placebo controlled clinical trial in Parkinson’s disease.Sci Rep 7:1398.
Martin-Bastida A,Lao-Kaim NP,Loane C,Politis M,Roussakis AA,Valle-Guzman N,Kefalopoulou Z,Paul-Visse G,Widner H,Xing Y,Schwarz ST,Auer DP,Foltynie T,Barker RA,Piccini P(2017b)Motor associations of iron accumulation in deep grey matter nuclei in Parkinson’s disease:a cross-sectional study of iron-related magnetic resonance imaging susceptibility.Eur J Neurol 24:357-365.
Monti DA,Zabrecky G,Kremens D,Liang TW,Wintering NA,Cai J,Wei X,Bazzan AJ,Zhong L,Bowen B,Intenzo CM,Iacovitti L,Newberg AB(2016)N-acetyl cysteine may support dopamine neurons in Parkinson’s disease:preliminary clinical and cell line data.PLoS One11:e0157602.
Nottingham SA,Springer JE(2003)Temporal and spatial distribution of activated caspase-3 after subdural kainic acid infusions in rat spinal cord.J Comp Neurol 464:463-471.
Ozawa H,Keane RW,Marcillo AE,Diaz PH,Dietrich WD(2002)Therapeutic strategies targeting caspase inhibition following spinal cord injury in rats.Exp Neurol 177:306-313.
Paterniti I,Mazzon E,Emanuela E,Paola RD,Galuppo M,Bramanti P,Cuzzocrea S(2010),Modulation of inflammatory response after spinal cord trauma with deferoxamine,an iron chelator.Free Radic Res 44:694-709.
Ropper AE,Ropper AH(2017)Acute spinal cord compression.N Engl J Med 376:1358-1369.
Stockwell BR et al.(2017)Ferroptosis:a regulated cell death nexus linking metabolism,redox biology,and disease.Cell 171:273-285.
Wang D,Liang J,Zhang J,Liu S,Sun W(2014a)Mild hypothermia combined with a scaffold of NgR-silenced neural stem cells/Schwann cells to treat spinal cord injury.Neural Regen Res 9:2189-2196.
Wang Y,Wang H,Tao Y,Zhang S,Wang J,Feng X(2014b)Necroptosis inhibitor necrostatin-1 promotes cell protection and physiological function in traumatic spinal cord injury.Neuroscience 266:91-101.
Watkins PA,Maiguel D,Jia Z,Pevsner J(2007)Evidence for 26 distinct acyl-coenzyme A synthetase genes in the human genome.J Lipid Res48:2736-2750.
Wu Q,Li YL,Ning GZ,Feng SQ,Chu TC,Li Y,Hao Y,Wu QL(2012)Epidemiology of traumatic cervical spinal cord injury in Tianjin,China.Spinal Cord 50:740-744.
Xiao W,Yu A,Liu D,Shen J,Xu Z(2015)Ligustilide treatment promotes functional recovery in a rat model of spinal cord injury via preventing ROS production.Int J Clin Exp Pathol 8:12005-12013.
Yu J,Guo Y,Sun M,Li B,Zhang Y,Li C(2009)Iron is a potential key mediator of glutamate excitotoxicity in spinal cord motor neurons.Brain Res 1257:102-107.
Zille M,Karuppagounder SS,Chen Y,Gough PJ,Bertin J,Finger J,Milner TA,Jonas EA,Ratan RR(2017)Neuronal death after hemorrhagic stroke in vitro and in vivo shares features of ferroptosis and necroptosis.Stroke 48:1033-1043.
Basso DM,Beattie MS,Bresnahan JC(1995)A sensitive and reliable locomotor rating scale for open field testing in rats.J Neurotrauma12:1-21.
Bazinet RP,Laye S(2014)Polyunsaturated fatty acids and their metabolites in brain function and disease.Nat Rev Neurosci 15:771-785.
Cakir O,Erdem K,Oruc A,Kilinc N,Eren N(2003)Neuroprotective effect of N-acetylcysteine and hypothermia on the spinal cord ischemia-reperfusion injury.Cardiovasc Surg 11:375-379.
Chen L,Hambright WS,Na R,Ran Q(2015)Ablation of the ferroptosis inhibitor glutathione peroxidase 4 in neurons results in rapid motor neuron degeneration and paralysis.J Biol Chem 290:28097-28106.
Choi DC,Lee JY,Lim EJ,Baik HH,Oh TH,Yune TY(2012)Inhibition of ROS-induced p38MAPK and ERK activation in microglia by acupuncture relieves neuropathic pain after spinal cord injury in rats.Exp Neurol 236:268-282.
Cozza G,Rossetto M,Bosello-Travain V,Maiorino M,Roveri A,Toppo S,Zaccarin M,Zennaro L,Ursini F(2017)Glutathione peroxidase 4-catalyzed reduction of lipid hydroperoxides in membranes:the polar head of membrane phospholipids binds the enzyme and addresses the fatty acid hydroperoxide group toward the redox center.Free Radic Biol Med 112:1-11.
Dixon SJ,Lemberg KM,Lamprecht MR,Skouta R,Zaitsev EM,Gleason CE,Patel DN,Bauer AJ,Cantley AM,Yang WS,Morrison B3rd,Stockwell BR(2012)Ferroptosis:an iron-dependent form of nonapoptotic cell death.Cell 149:1060-1072.
Do Van B,Gouel F,Jonneaux A,Timmerman K,Gele P,Petrault M,Bastide M,Laloux C,Moreau C,Bordet R,Devos D,Devedjian JC(2016)Ferroptosis,a newly characterized form of cell death in Parkinson’s disease that is regulated by PKC.Neurobiol Dis 94:169-178.
Fan B,Wei Z,Yao X,Shi G,Cheng X,Zhou X,Zhou H,Ning G,Kong X,Feng S(2018)Microenvironment imbalance of spinal cord injury.Cell Transplant 27:853-866.
Fan H,Zhang K,Shan L,Kuang F,Chen K,Zhu K,Ma H,Ju G,Wang YZ(2016)Reactive astrocytes undergo M1 microglia/macrohpages-induced necroptosis in spinal cord injury.Mol Neurodegener11:14.
Genovese T,Mazzon E,Esposito E,Muia C,Di Paola R,Di Bella P,Bramanti P,Cuzzocrea S(2007)Role of endogenous glutathione in the secondary damage in experimental spinal cord injury in mice.Neurosci Lett 423:41-46.
Girotti AW(1998)Lipid hydroperoxide generation,turnover,and effector action in biological systems.J Lipid Res 39:1529-1542.
Grossman SD,Rosenberg LJ,Wrathall JR(2001a)Relationship of altered glutamate receptor subunit mRNA expression to acute cell loss after spinal cord contusion.Exp Neurol 168:283-289.
Grossman SD,Rosenberg LJ,Wrathall JR(2001b)Temporal-spatial pattern of acute neuronal and glial loss after spinal cord contusion.Exp Neurol 168:273-282.
Guiney SJ,Adlard PA,Bush AI,Finkelstein DI,Ayton S(2017)Ferroptosis and cell death mechanisms in Parkinson’s disease.Neurochem Int 104:34-48.
Hao J,Li B,Duan HQ,Zhao CX,Zhang Y,Sun C,Pan B,Liu C,Kong XH,Yao X,Feng SQ(2017)Mechanisms underlying the promotion of functional recovery by deferoxamine after spinal cord injury in rats.Neural Regen Res 12:959-968.
He M,Ding Y,Chu C,Tang J,Xiao Q,Luo ZG(2016)Autophagy induction stabilizes microtubules and promotes axon regeneration after spinal cord injury.Proc Natl Acad Sci U S A 113:11324-11329.
Huang H,Young W,Ziad A,Hooshang S,Alok S,Dafin M,Shiqing F,Lin C(2015)Beijing declaration of international association of neurorestoratology.J Neurorestoratol 3:121-122.
Imai H,Matsuoka M,Kumagai T,Sakamoto T,Koumura T(2017)Lipid peroxidation-dependent cell death regulated by GPx4 and ferroptosis.Curr Top Microbiol Immunol 403:143-170.
Koozekanani SH,Vise WM,Hashemi RM,McGhee RB(1976)Possible mechanisms for observed pathophysiological variability in experimental spinal cord injury by the method of Allen.J Neurosurg44:429-434.
Li Q,Han X,Lan X,Gao Y,Wan J,Durham F,Cheng T,Yang J,et al.(2017),Inhibition of neuronal ferroptosis protects hemorrhagic brain.JCI Insight 2:e90777.
Liu JM,Wang FC,Zhou YJ,Mu L,Hou SK,Hao LN,Zhang ZT(2016)Effect of electroacupuncture stimulation on apoptosis of nerve cells in a rat model of spinal cord contusion.Zhongguo Zuzhi Gongcheng Yanjiu 20:616-621.
Liu XZ,Xu XM,Hu R,Du C,Zhang SX,McDonald JW,Dong HX,Wu YJ,Fan GS,Jacquin MF,Hsu CY,Choi DW(1997)Neuronal and glial apoptosis after traumatic spinal cord injury.J Neurosci 17:5395-5406.
Livak KJ,Schmittgen TD(2001)Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T))Method.Methods 25:402-408.
Lu J,Ashwell KW,Waite P(2000)Advances in secondary spinal cord injury:role of apoptosis.Spine(Phila Pa 1976)25:1859-1866.
Martin-Bastida A,Ward RJ,Newbould R,Piccini P,Sharp D,Kabba C,Patel MC,Spino M,Connelly J,Tricta F,Crichton RR,Dexter DT(2017a)Brain iron chelation by deferiprone in a phase 2 randomised double-blinded placebo controlled clinical trial in Parkinson’s disease.Sci Rep 7:1398.
Martin-Bastida A,Lao-Kaim NP,Loane C,Politis M,Roussakis AA,Valle-Guzman N,Kefalopoulou Z,Paul-Visse G,Widner H,Xing Y,Schwarz ST,Auer DP,Foltynie T,Barker RA,Piccini P(2017b)Motor associations of iron accumulation in deep grey matter nuclei in Parkinson’s disease:a cross-sectional study of iron-related magnetic resonance imaging susceptibility.Eur J Neurol 24:357-365.
Monti DA,Zabrecky G,Kremens D,Liang TW,Wintering NA,Cai J,Wei X,Bazzan AJ,Zhong L,Bowen B,Intenzo CM,Iacovitti L,Newberg AB(2016)N-acetyl cysteine may support dopamine neurons in Parkinson’s disease:preliminary clinical and cell line data.PLoS One11:e0157602.
Nottingham SA,Springer JE(2003)Temporal and spatial distribution of activated caspase-3 after subdural kainic acid infusions in rat spinal cord.J Comp Neurol 464:463-471.
Ozawa H,Keane RW,Marcillo AE,Diaz PH,Dietrich WD(2002)Therapeutic strategies targeting caspase inhibition following spinal cord injury in rats.Exp Neurol 177:306-313.
Paterniti I,Mazzon E,Emanuela E,Paola RD,Galuppo M,Bramanti P,Cuzzocrea S(2010),Modulation of inflammatory response after spinal cord trauma with deferoxamine,an iron chelator.Free Radic Res 44:694-709.
Ropper AE,Ropper AH(2017)Acute spinal cord compression.N Engl J Med 376:1358-1369.
Stockwell BR et al.(2017)Ferroptosis:a regulated cell death nexus linking metabolism,redox biology,and disease.Cell 171:273-285.
Wang D,Liang J,Zhang J,Liu S,Sun W(2014a)Mild hypothermia combined with a scaffold of NgR-silenced neural stem cells/Schwann cells to treat spinal cord injury.Neural Regen Res 9:2189-2196.
Wang Y,Wang H,Tao Y,Zhang S,Wang J,Feng X(2014b)Necroptosis inhibitor necrostatin-1 promotes cell protection and physiological function in traumatic spinal cord injury.Neuroscience 266:91-101.
Watkins PA,Maiguel D,Jia Z,Pevsner J(2007)Evidence for 26 distinct acyl-coenzyme A synthetase genes in the human genome.J Lipid Res48:2736-2750.
Wu Q,Li YL,Ning GZ,Feng SQ,Chu TC,Li Y,Hao Y,Wu QL(2012)Epidemiology of traumatic cervical spinal cord injury in Tianjin,China.Spinal Cord 50:740-744.
Xiao W,Yu A,Liu D,Shen J,Xu Z(2015)Ligustilide treatment promotes functional recovery in a rat model of spinal cord injury via preventing ROS production.Int J Clin Exp Pathol 8:12005-12013.
Yu J,Guo Y,Sun M,Li B,Zhang Y,Li C(2009)Iron is a potential key mediator of glutamate excitotoxicity in spinal cord motor neurons.Brain Res 1257:102-107.
Zille M,Karuppagounder SS,Chen Y,Gough PJ,Bertin J,Finger J,Milner TA,Jonas EA,Ratan RR(2017)Neuronal death after hemorrhagic stroke in vitro and in vivo shares features of ferroptosis and necroptosis.Stroke 48:1033-1043.