脊髓损伤后虫草素增强突触分化诱导基因1的表达
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摘要
观察脊髓损伤后虫草素对脊髓组织内突触分化诱导基因1(SynDIG1)表达和神经系统运动功能的影响,探讨神经损伤修复的内在机制。将成年雌性SD大鼠随机分为对照组、损伤组、低剂量和高剂量治疗组,对照组不损伤脊髓,治疗组与损伤组行脊髓半横切损伤,随后治疗组给予虫草素处理,损伤组给予生理盐水处理。术后14、21、28、35 d用BBB和斜板试验评价大鼠肢体运动功能,并用免疫荧光和免疫印迹测定脊髓组织内SynDIG1活性。免疫荧光和免疫印迹结果均表明术后14 d损伤组和治疗组均有神经细胞SynDIG1表达,在28 d达到高峰,之后呈高水平表达,且SynDIG1水平在高剂量治疗组明显高于低剂量治疗组和损伤组,BBB和斜板试验评分在高剂量治疗组均明显高于低剂量治疗组和损伤组。虫草素明显改善损伤区域的微环境,激活神经系统的内源性修复机制,促进神经细胞SynDIG1大量表达,有利于神经系统运动功能恢复。
Objective : To observe the effect of cordycepin on synapse differentiation induced gene 1(SynDIGl) expression and the motor function of the nervous system in spinal cord injury, and to explore the internal mechanism of nerve injury repair.Methods : SD rats were randomly divided into control group, injury group, low and high dose treatment group. The injury and treatment groups were injured by spinal cord hemisection. The control group had no spinal cord injury. The injury and treatment groups were respectively injected with cordycepin and normal saline. At 14,21,28,35 d,the motor neural function was evaluated by inclined plane test and Basso Beattie Bresnahan score(BBB). Immunofluorescence staining and Western blotting were used to observe SynDIGl expression. Results: Western blotting and immunofluorescence staining results showed that the expression of SynDIGl in injury and treatment group were found at 14 d after the spinal cord injury,and reached the peak at 28 d after the injury. The expression of SynDIGl of high-dose group was significantly higher than that of the low-dose group and injury group. BBB and inclined plane test score of high-dose group were significantly higher than that of the low-dose group and injury group. Conclusion: Cordycepin can significantly improve the microenvironment of the injury area, activate the endogenous recovery mechanism of the nervous system and promote the expression of SynDIGl. It is beneficial for the recovery of motor functions of the nervous system.
Objective : To observe the effect of cordycepin on synapse differentiation induced gene 1(SynDIGl) expression and the motor function of the nervous system in spinal cord injury, and to explore the internal mechanism of nerve injury repair.Methods : SD rats were randomly divided into control group, injury group, low and high dose treatment group. The injury and treatment groups were injured by spinal cord hemisection. The control group had no spinal cord injury. The injury and treatment groups were respectively injected with cordycepin and normal saline. At 14,21,28,35 d,the motor neural function was evaluated by inclined plane test and Basso Beattie Bresnahan score(BBB). Immunofluorescence staining and Western blotting were used to observe SynDIGl expression. Results: Western blotting and immunofluorescence staining results showed that the expression of SynDIGl in injury and treatment group were found at 14 d after the spinal cord injury,and reached the peak at 28 d after the injury. The expression of SynDIGl of high-dose group was significantly higher than that of the low-dose group and injury group. BBB and inclined plane test score of high-dose group were significantly higher than that of the low-dose group and injury group. Conclusion: Cordycepin can significantly improve the microenvironment of the injury area, activate the endogenous recovery mechanism of the nervous system and promote the expression of SynDIGl. It is beneficial for the recovery of motor functions of the nervous system.
引文
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[2]马延超.ERK1/2信号通路调控成年大鼠脊髓损伤后谷氨酸转运体GLAST和GLT-1表达的实验研究[D].兰州:兰州大学,2014:1.
[3]Tuli H S,Sharma A K,Sandhu S S,et al.Cordycepin:a bioactive metabolite with therapeutic potential[J].Life Sci,2013,93(23):863-869.
[4]张姝,张永杰,ShresthaB,等.冬虫夏草菌和蛹虫草菌的研究现状、问题及展望[J].菌物学报,2013,32(4):577-597.
[5]赵富生,张晓莉,武庚,等.大鼠两种脊髓半切模型制备方法的改良及比较[J].实验动物与比较医学,2014,34(1):6-11.
[6]刘小康,徐建广,连小峰,等.大鼠钳夹式急性脊髓损伤模型的制备与评价[J].中国矫形外科杂志,2012,20(14):1318-1322.
[7]Wang B,Li Y,Li X P,et al.Panax notoginseng saponins improve recovery after spinal cord transection by up regulating neurotrophic factors[J].Neural Regen Res,2015,10(8):1317-1320.
[8]Ropper A E,Zeng X,Anderson J E,et al.An efficient device to experimentally model compression injury of mammalian spinal cord[J].Exp Neurol,2015,271:515-523.
[9]Mensch S,Baraban M,Almeida R,et al.Synaptic vesicle release regulates myelin sheath number of individual oligodendrocytes in vivo[J].Nat Neurosci,2015,18(5):628-630.
[10]Kitamura M,Kato H,Saito Y,et al.Aberrant,differential and bidirectional regulation of the unfolded protein response towards cell survival by 3'-deoxyadenosine[J].Cell Death Differ,2011,18(12):1876-1888.
[11]Pardridge W M.Drug transport across the blood-brain barrier[J].J Cereb Blood Flow Metab,2012,32(11):1959-1972.
[12]Ye X H,Wu Y,Guo P P,et al.Lipoxin A4 analogue protects brain and reduces inflammation in a rat model of focal cerebral ischemia reperfusion[J].Brain Res,2010,1323:174-183.
[13]Cai Z L,Wang C Y,Jiang Z J,et al.Effects of cordycepin on Ymaze learning task in mice[J].Eur J Pharmacol,2013,714(1-3):249-253.
[14]Peng J,Wang P,Ge H,et al.Effects of cordycepin on the microglia-overactivation-induced impairments of growth and development of hippocampal cultured neurons[J].PLoS One,2015,10(5):e0125902.
[15]Aruna Devi R,Lata S,Bhadoria B K,et al.Neuro protective effect of 5,7,3',4',5'-pentahydroxydihydroflavanol-3-O-(2"-Ogalloyl)-beta-D-glucopyranoside,a polyphenolic compound in focal cerebral ischemia in rat[J].Eur J Pharmacol,2010,626(2-3):205-212.
[16]Cheng Z,He W,Zhou X,et al.Cordycepin protects against cerebral ischemia/reperfusion injury in vivo and in vitro[J].Eur J Pharmacol,2011,664(1-3):20-28.
[17]Ramesh T,Yoo S K,Kim S W,et al.Cordycepin(3'-deoxyadenosine)attenuates age-related oxidative stress and ameliorates antioxidant capacity in rats[J].Exp Gerontol,2012,47(12):979-987.
[18]Nunez-Figueredo Y,Garcia-Pupo L,Ramirez-Sanchez J,et al.Neuroprotective action and free radical scavenging activity of Guttiferone-A,a naturally occurring prenylated benzophenone[J].Arzneimittelforschung,2012,62(12):583-589.
[19]李巍.谷氨酸受体相互作用蛋白介导AMPA受体兴奋性毒性效应[D].西安:第四军医大学,2009:87.
[20]Carlezon W A Jr,Nestler E J.Elevated levels of GluR1 in the midbrain:a trigger for sensitization to drugs of abuse?[J].Trends Neurosci,2002,25(12):610-615.
[21]Fredholm B B,IJzerman A P,Jacobson K A,et al.International Union of Basic and Clinical Pharmacology.LXXXI.Nomenclature and classification of adenosine receptors an update[J].Pharmacology,2011,63(1):1-34.
[22]Du J,Creson TK,Wu LJ,et al.The role of hippocampal GluR1and GluR2 receptors in manic-like behavior[J].J Neurosci,2008,28(1):68-79.
[23]Du J,Suzuki K,Wei Y,et al.The anticonvulsants lamotrigine,riluzole,and valproate differentially regulate AMP A receptor membrane localization:relationship to clinical effects in mood disorders[J].Neuropsychopharmacology,2007,32(4):793-802.