成年大鼠视神经夹伤后TLR4与GAP-43表达时程的比较
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摘要
目的:明确视神经损伤引起的固有免疫应答反应与视网膜神经节细胞轴突再生之间的时间关系。方法:将成年SD大鼠随机分为视神经损伤组与假手术对照组,损伤组动物左侧视神经以钳夹法损伤,对照组仅暴露左侧视神经。手术第1、3、7 d处死后取左侧视神经干,用Western Blot方法检测视神经干toll样受体4(toll-like receptor 4,TLR4)和生长相关蛋白-43(growth-associated protein-43,GAP-43)蛋白表达水平。结果:视神经干内TLR4和GAP-43蛋白在损伤后第1 d都趋于表达上调,但其后的变化趋势具有不同的时间特征,即TLR4持续上调,并在损伤后7~14 d内维持呈平台;而GAP-43的表达在损伤后第7 d呈峰值,其后逐渐下降,第14 d基本恢复至基线水平。结论:TLR4介导的固有免疫应答有可能影响视神经再生;TLR4和GAP-43表达变化的不同为进一步研究固有免疫应答对视神经再生的影响提供了有益的线索。
Objective: To determine the relationship of time course between injury-induced innate immune reaction and axon regeneration of retinal ganglion cells after optic nerve( ON) crush in adult rats. Methods: Adult SD rats were divided into ON injury group and sham operation group. The left ON was crushed or only exposed for the injury group or the sham operation group,respectively. Then the rats were sacrificed at 1,3 and 7 d after operation,and the protein levels of toll-like receptor 4( TLR4) and growth-associated protein-43( GAP-43) in the left ON were detected with Western Blot assay. Results: Both TLR4 and GAP-43 were upregulated after ON crush,but their expression changed in different pattern. Namely,TLR4 expression was increased consistently and keep stably within 7 ~ 14 d; while GAP-43protein level peaked at 7 d,then dropped back to the baseline at 14 d. Conclusion: TLR4 mediated innate immune reaction may influence the ON regeneration. This study thus provided a useful clue for further studies in the effects of the innate immune reaction on the ON regeneration.
Objective: To determine the relationship of time course between injury-induced innate immune reaction and axon regeneration of retinal ganglion cells after optic nerve( ON) crush in adult rats. Methods: Adult SD rats were divided into ON injury group and sham operation group. The left ON was crushed or only exposed for the injury group or the sham operation group,respectively. Then the rats were sacrificed at 1,3 and 7 d after operation,and the protein levels of toll-like receptor 4( TLR4) and growth-associated protein-43( GAP-43) in the left ON were detected with Western Blot assay. Results: Both TLR4 and GAP-43 were upregulated after ON crush,but their expression changed in different pattern. Namely,TLR4 expression was increased consistently and keep stably within 7 ~ 14 d; while GAP-43protein level peaked at 7 d,then dropped back to the baseline at 14 d. Conclusion: TLR4 mediated innate immune reaction may influence the ON regeneration. This study thus provided a useful clue for further studies in the effects of the innate immune reaction on the ON regeneration.
引文
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[6]Benowitz LI,Routtenberg A.GAP-43:an intrinsic determinant of neuronal development and plasticity[J].Trends Neurosci,1997,20:84-91.
[7]Kaneda M,Nagashima M,Mawatari K,et al.Growth-associated protein43(GAP43)is a biochemical marker for the whole period of fish optic nerve regeneration[J].AdvExp Med Biol,2010,664:97-104.
[8]Kriz J.Inflammation in ischemic brain injury:timing is important[J].Crit Rev Neurobiol,2006,18:145-157.
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[10]刁金美,冯国栋,游思维,等.用于中枢神经系统Waller变性研究的新型大鼠视神经夹伤模型[J].神经解剖学杂志,2008,24:239-244.
[11]Nitzan A,Kermer P,Shirvan A,et al.AS.Examination of cellular and molecular events associated with optic nerve axotomy[J].Glia,2006,54:545-556.
[12]Yao AH,Jia LY,Zhang YK,et al.Early blockade of TLRs MyD88-dependent pathway may reduce secondary spinal cord injury in the rats[J].Evid Based Complement Alternat Med,2012,591298.
[13]Zhang YK,Liu JT,Peng ZW,et al.Different TLR4 expression and microglia/macrophage activation induced by hemorrhage in the rat spinal cord after compressiveinjury[J].J Inflam,2013,10:112.
[2]Griffiths MR,Gasque P,Neal JW.The regulation of the CNS innate immune response is vital for the restoration of tissue homeostasis(repair)after acute brain injury:a brief review[J].Int Inflam,2010,2010:151097.
[3]Lehnardt S.Innate immunity and neuroinflammation in the CNS:the role of microglia in Toll-like receptor-mediated neuronal injury[J].Glia,2010,58:253-263.
[4]vanNoort JM,Bsibsi M.Toll-like receptors in the CNS:Implications for neurodegeneration and repair[J].Prog Brain Res,2009,175:139-148.
[5]Zheng Z,Yuan R,Song M,et al.The toll-like receptor 4-mediated signaling pathway is activated following optic nerve injury in mice[J].Brain Res,2012,1489:90-97.
[6]Benowitz LI,Routtenberg A.GAP-43:an intrinsic determinant of neuronal development and plasticity[J].Trends Neurosci,1997,20:84-91.
[7]Kaneda M,Nagashima M,Mawatari K,et al.Growth-associated protein43(GAP43)is a biochemical marker for the whole period of fish optic nerve regeneration[J].AdvExp Med Biol,2010,664:97-104.
[8]Kriz J.Inflammation in ischemic brain injury:timing is important[J].Crit Rev Neurobiol,2006,18:145-157.
[9]Schwartz M.Optic nerve crush:protection and regeneration[J].Brain Res Bull,2004,62:467-471.
[10]刁金美,冯国栋,游思维,等.用于中枢神经系统Waller变性研究的新型大鼠视神经夹伤模型[J].神经解剖学杂志,2008,24:239-244.
[11]Nitzan A,Kermer P,Shirvan A,et al.AS.Examination of cellular and molecular events associated with optic nerve axotomy[J].Glia,2006,54:545-556.
[12]Yao AH,Jia LY,Zhang YK,et al.Early blockade of TLRs MyD88-dependent pathway may reduce secondary spinal cord injury in the rats[J].Evid Based Complement Alternat Med,2012,591298.
[13]Zhang YK,Liu JT,Peng ZW,et al.Different TLR4 expression and microglia/macrophage activation induced by hemorrhage in the rat spinal cord after compressiveinjury[J].J Inflam,2013,10:112.