SD大鼠脊髓损伤后微环境中髓鞘相关抑制因子的表达
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摘要
目的观察SD大鼠在不同致伤势能的Allen’s模型和脊髓全切模型微环境中不同时间点的髓鞘相关抑制因子表达。方法选择125只成年雌性SD大鼠随机分为5组,每组25只, A组(假手术组), B组[20 g×5 cm,即致伤势能100 gcf (gram-cm force)], C组(20 g×10 cm,致伤势能200 gcf), D组(20 g×15 cm,致伤势能300 gcf)和E组(脊髓全切组)。采用Western blotting印迹分析法,在制模成功后1 d、5 d、7 d、14 d、28 d观察大鼠脊髓损伤(SCI)组织中勿动蛋白-A(Nogo-A)、髓鞘相关糖蛋白(MAG)、少突胶质细胞髓鞘糖蛋白(OMgp)等髓鞘相关抑制因子表达。结果整个实验中大鼠死亡24只(16.67%); B、C、D、E组SCI后1 d可见Nogo-A、MAG、OMgp表达,Nogo-A与MAG表达于SCI后7 d达峰值,28 d恢复到假手术组水平; SCI后1 d可见髓鞘中OMgp表达,5 d达峰值,28 d恢复至假手术组水平。结论 Nogo-A、MAG、OMgp在SCI后普遍呈现高表达,并在SCI后5～7 d达到峰值。
Objective To observe the expression of myelin related inhibitory factor in SpragueDawley (SD) rats at different time points in different injury potential energy of Allen's model and spinal cord total cut model with the microenvironment. Methods One hundred and twenty five adult female healthy SD rats were selected and randomly divided into 5 groups with 25 in each, group A (sham operation group), group B [(20 g × 5 cm, injury potential energy 100 gram-cm force (gfc))], group C (20 g × 10 cm, injury potential energy 200 gcf), group D (20 g × 15 cm, injury potential energy 300 gcf); group E(total spinal cord resection group).The expression of myelin related inhibitory factors of neurite outgrowth inhibitor-A(Nogo-A), the myelin associated glycoprotein(MAG), and the oligodendrocyte myelin glycoprotein (OMgp) were analyzed by Western blotting after spinal cord injury (SCI) at 1 d, 5 d, 7 d, 14 d and 28 d after successful modeling. Results Twenty-four cases(16.67%) of SD rats were died in the whole experiment. The expression of Nogo-A, MAG and OMgp was observed at the first day after SCI in group B, C, D and E. The expression of Nogo-A and MAG reached the peak at the 7th day and returned to the levels of sham operation group on the 28th day. The expression of OMgp in myelin was observed at the first day after SCI, and reached the peak at day 5, and returned to the levels of sham operation group on the 28th day. Conclusion The Nogo-A, MAG and OMgp are highly expressed after SCI and those indexes are reached the peak 5～7 days after SCI.
Objective To observe the expression of myelin related inhibitory factor in SpragueDawley (SD) rats at different time points in different injury potential energy of Allen's model and spinal cord total cut model with the microenvironment. Methods One hundred and twenty five adult female healthy SD rats were selected and randomly divided into 5 groups with 25 in each, group A (sham operation group), group B [(20 g × 5 cm, injury potential energy 100 gram-cm force (gfc))], group C (20 g × 10 cm, injury potential energy 200 gcf), group D (20 g × 15 cm, injury potential energy 300 gcf); group E(total spinal cord resection group).The expression of myelin related inhibitory factors of neurite outgrowth inhibitor-A(Nogo-A), the myelin associated glycoprotein(MAG), and the oligodendrocyte myelin glycoprotein (OMgp) were analyzed by Western blotting after spinal cord injury (SCI) at 1 d, 5 d, 7 d, 14 d and 28 d after successful modeling. Results Twenty-four cases(16.67%) of SD rats were died in the whole experiment. The expression of Nogo-A, MAG and OMgp was observed at the first day after SCI in group B, C, D and E. The expression of Nogo-A and MAG reached the peak at the 7th day and returned to the levels of sham operation group on the 28th day. The expression of OMgp in myelin was observed at the first day after SCI, and reached the peak at day 5, and returned to the levels of sham operation group on the 28th day. Conclusion The Nogo-A, MAG and OMgp are highly expressed after SCI and those indexes are reached the peak 5～7 days after SCI.
引文
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[13]赵富生,武庚,武杨,等.大鼠脊髓楔型半切空洞模型的建立及评估[J].实验动物科学,2013,30(4):13-17.
[14]陈斌,武庚,黄喆,等.大鼠脊髓不同平面双侧半横切损伤模型的建立及评价[J].实验动物科学,2015,32(1):7-11.
[15]Cafferty WB,Duffy P,Huebner E,et al.MAG and OMgp synergize with Nogo-A to restrict axonal growth and neurological recovery after spinal cord trauma[J].J Neurosci,2010,19(30):6825-6837
[16]Wang KC,Koprivica V,Kin TA,et al.Oligodendrocytemyeling lycoprotein is a Nogo recept or ligand that in hibits neurite out growth[J].Nature,2002,417(6892):941-944.
[17]Kottis V,Thibault P,Mikol D,et al.Oligod end rocy temyelin glycoprotein(OMgp)is an inhibitor of neurite out growth[J].J Neurochem,2002,82(6):1566-1569.
[18]Gao L,Macklin W,Gerson J.Intrinsic and extrinsic inhibition of oligoden-drocyte development by rat retina[J].Dev Biol,2006,(2):277-286.
[19]Leibinger M,Müller A,Gobrecht P,et al.Interleukin-6contributes to CNS axon regeneration upon inflammatory stimulation[J].Cell Death Dis,2013,4(4):e609.
[20]张大威,李一帆,朱丹.急性大鼠脊髓损伤模型的建立与评估[J].中风与神经疾病杂志,2015,32(4):325-327.
[2]Mckerracher L,Ferraro GB,Fournier AE.Chapter Six-Rho signaling and axon regeneration[J].Int Rev Neurobiol,2012,105(1):117-140.
[3]崔翔,马冉,唐萁,等.针灸对AD模型大鼠海马轴突生长抑制因子MAG及OMgp表达的影响[J].时珍国医国药,2014,25(7):1752-1754.
[4]Lim SN,Huang W,Hall JCE,et al.Improved outcome after spinal cord compression injury in mice treated with docosahexaenoic acid[J].Exp Neurol,2013,239(1):13-27.
[5]Tan HB,Zhong YS,Cheng Y,et al.Rho/ROCK pathway and neural regeneration:a potential therapeutic target for central nervous system and optic nerve damage[J].Int J Ophthalmol,2011,4(6):652-657.
[6]胡怀强,周永红,马学盛,等.复健片对大鼠脑梗死后不同时间点Nogo-A表达的影响[J].中国实验动物学报,2010,18(2):118-121.
[7]Teo L,Rosenfeld JV,Bourne JA.Models of CNS injury in the nonhuman primate:A new era for treatment strategies[J].Transl Neuroscl,2012,3(2):181-195.
[8]杨俊锋,张亚峰,马勇,等.Nogo-A在脊髓损伤大鼠脊髓组织中的动态表达[J].中国康复医学杂志,2015,30(9):867-871.
[9]马睿杰,张柳娟,孙连珠,等.夹脊电针干预脊髓损伤大鼠OMgp表达的研究[J].中华中医药学刊,2014,17(6):1283-1286.
[10]Zhai P,Chen XB,Schreyer DJ.An in vitro study of peptideloaded alginate nanospheres for antagonizing the inhibitory effect of Nogo-A protein on axonal growth[J].Biomed Mater,2015,10(4):045016.
[11]Mullner A,Gonzenbach R,Weinmann R,et al.Laminaspecific restoration of serotonergic projections after Nogo-Aantibody treatment of spinal cord injury in rats[J].Eur JNeurosci,2008,27(2)326-333.
[12]Kinter J,Lazzati T,Schmid D,et al.An essential role of MAGin mediating axon-myelin attachment in Charcot-Marie-Tooth1A disease[J].Neurobiol Dis,2013,49(1):221-231.
[13]赵富生,武庚,武杨,等.大鼠脊髓楔型半切空洞模型的建立及评估[J].实验动物科学,2013,30(4):13-17.
[14]陈斌,武庚,黄喆,等.大鼠脊髓不同平面双侧半横切损伤模型的建立及评价[J].实验动物科学,2015,32(1):7-11.
[15]Cafferty WB,Duffy P,Huebner E,et al.MAG and OMgp synergize with Nogo-A to restrict axonal growth and neurological recovery after spinal cord trauma[J].J Neurosci,2010,19(30):6825-6837
[16]Wang KC,Koprivica V,Kin TA,et al.Oligodendrocytemyeling lycoprotein is a Nogo recept or ligand that in hibits neurite out growth[J].Nature,2002,417(6892):941-944.
[17]Kottis V,Thibault P,Mikol D,et al.Oligod end rocy temyelin glycoprotein(OMgp)is an inhibitor of neurite out growth[J].J Neurochem,2002,82(6):1566-1569.
[18]Gao L,Macklin W,Gerson J.Intrinsic and extrinsic inhibition of oligoden-drocyte development by rat retina[J].Dev Biol,2006,(2):277-286.
[19]Leibinger M,Müller A,Gobrecht P,et al.Interleukin-6contributes to CNS axon regeneration upon inflammatory stimulation[J].Cell Death Dis,2013,4(4):e609.
[20]张大威,李一帆,朱丹.急性大鼠脊髓损伤模型的建立与评估[J].中风与神经疾病杂志,2015,32(4):325-327.