骨形态发生蛋白7促进大鼠急性脊髓损伤后神经功能修复的研究
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摘要
目的探讨骨形态发生蛋白7(bone morphogenetic protein 7,BMP7)是否具有促进大鼠急性脊髓损伤后神经功能修复的作用。方法清洁级Wistar大鼠分为假手术组(SO组)、阴性对照组(NC组)和BMP 7实验组。假手术组仅暴露第10胸椎硬脊膜,但并不对其进行损伤。NC组以Allences′s打击器进行第10胸椎脊髓损伤模型,造模成功后,每天给第10胸椎硬脊膜处注射生理盐水,连续7天。BMP7组造模成功后,每天给损伤处注射BMP 7蛋白50 ng,连续7天。各组分别于术后6小时、3天、1周、2周、4周和8周进行BBB量表评分及利用Western blot检测损伤节段脊髓神经丝蛋白200 (neurofilament protein200, NF200)及胶质纤微酸性蛋白(glial fibrillary acidic protein,GFAP)表达量的变化,于术后1周和8周采用生物信号采集和处理系统检测运动诱发电位(MEP)。结果 BBB评分结果显示SO组后肢运动功能基本正常。NC组和BMP 7组后肢运动功能在术后完全丧失,3天后逐渐恢复,在1周、2周、4周和8周,BMP 7组的BBB评分均高于NC组(P <0.05)。神经电生理测试结果显示,SO组胸10水平的MEP波形为正常,即P1-N1-P2。术后1周NC组和BMP 7组MEP波N1波均低于正常值,术后8周BMP 7组和NC组以M形双峰波为主,且前者波幅高于后者,但其较术后1周都明显增高。WesternBlot结果显示,NC组NF 200的表达一直无变化,BMP7组NF 200表达在术后3天明显升高,且持续增长直至至第4周;NC组和BMP 7组GFAP表达水平在2w达到高峰,随后下降,两组间无显著差异。结论 BMP7蛋白具有促进大鼠脊髓损伤后神经功能修复的作用。
Objective To investigate whether BMP7 plays a role in the repair of nerve function after acute spinal cord injury in rats. Methods The Wistar of clean grade rats were divided into the sham operation group(SO group), the negative control group(NC group)and the BMP7 experimental group. The SO group only exposed the 10 th thoracic vertebra endorachis, but did not injure it. NC group with Allences' s cause 10 thoracic spinal cord injury model, and injected with physiological saline daily for 7 days after successful modeling. BMP7 group was injected with BMP-7 protein 50 ng daily for 7 days after successful modeling. The scores of BBB and the injured segmental spinal cord of levels of NF200 and GFAP were measured by Western blot at 6 hours, 3 days, 1 week, 2 weeks, 4 weeks and 8 weeks after operation in each group. The motor evoked potentials(MEP)were detected by biosignal acquisition and processing system at 1 week and 8 weeks after operation. Results The results of BBB score showed that motor function of hindlimb in SO group was normal. The hindlimb motor function of NC group and BMP 7 group was completely lost after operation and recovered gradually after 3 days. BBB score of BMP 7 group was higher than that of NC group(P < 0.05) at 1 week, 2 weeks, 4 weeks and 8 weeks. Nerveelectrophysiological test results showed that the MEP waveform at the thoracic level of SO group was normal, that is P1-N1-P2. N1 wave of MEP wave in NC group and BMP7 group was lower than that of normal value at 1 week after operation. M-shaped bimodal wave was dominant in BMP7 group and NC group at 8 weeks after operation, and the amplitude of former was higher than that of the latter,but they all significantly higher than 1 week after operation. The results of WesternBlot showed that the expression of NF 200 remained unchanged in NC group, while the expression of NF 200 in BMP7 group was significantly increased at 3 days after operation and continued to increase until the 4 th week. The GFAP expression in NC group and BMP 7 group peaked at 2 weeks and then decreased, but no significantly difference was found between the two groups. Conclusion BMP7 protein can promote the repair of nerve function after spinal cord injury in rats.
Objective To investigate whether BMP7 plays a role in the repair of nerve function after acute spinal cord injury in rats. Methods The Wistar of clean grade rats were divided into the sham operation group(SO group), the negative control group(NC group)and the BMP7 experimental group. The SO group only exposed the 10 th thoracic vertebra endorachis, but did not injure it. NC group with Allences' s cause 10 thoracic spinal cord injury model, and injected with physiological saline daily for 7 days after successful modeling. BMP7 group was injected with BMP-7 protein 50 ng daily for 7 days after successful modeling. The scores of BBB and the injured segmental spinal cord of levels of NF200 and GFAP were measured by Western blot at 6 hours, 3 days, 1 week, 2 weeks, 4 weeks and 8 weeks after operation in each group. The motor evoked potentials(MEP)were detected by biosignal acquisition and processing system at 1 week and 8 weeks after operation. Results The results of BBB score showed that motor function of hindlimb in SO group was normal. The hindlimb motor function of NC group and BMP 7 group was completely lost after operation and recovered gradually after 3 days. BBB score of BMP 7 group was higher than that of NC group(P < 0.05) at 1 week, 2 weeks, 4 weeks and 8 weeks. Nerveelectrophysiological test results showed that the MEP waveform at the thoracic level of SO group was normal, that is P1-N1-P2. N1 wave of MEP wave in NC group and BMP7 group was lower than that of normal value at 1 week after operation. M-shaped bimodal wave was dominant in BMP7 group and NC group at 8 weeks after operation, and the amplitude of former was higher than that of the latter,but they all significantly higher than 1 week after operation. The results of WesternBlot showed that the expression of NF 200 remained unchanged in NC group, while the expression of NF 200 in BMP7 group was significantly increased at 3 days after operation and continued to increase until the 4 th week. The GFAP expression in NC group and BMP 7 group peaked at 2 weeks and then decreased, but no significantly difference was found between the two groups. Conclusion BMP7 protein can promote the repair of nerve function after spinal cord injury in rats.
引文
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[14]Lam S K,Sayama C,Harris D A,et al.Nationwide practice patterns in the use of recombinant human bone morphogenetic protein-2 in pediatric spine surgery as a function of patient-,hospital-,and procedure-related factors[J].J Neurosurg Pediatr,2014,14(5):476-485.
[15]Tannoury C A,An H S.Complications with the use of bone morphogenetic protein 2(BMP-2)in spine surgery.[J].Spine J,2014,14(3):552.
[2]LF Rodrigues,V MouraNeto,TCLS Spohr E.Biomarkers in spinal cord injury:from prognosis to treatment[J].Mol Neurobiol,2018(2):1-13.
[3]Han D,Wu C,Xiong Q,et al.Anti-inflammatory mechanism of bone marrow mesenchymal stem cell transplantation in rat model of spinal cord injury[J].Cell Biochem Biophys,2015,71(3):1341-1347.
[4]Yang W,Yang Y,Yang J Y,et al.Treatment with bone marrow mesenchymal stem cells combined with plumbagin alleviates spinal cord injury by affecting oxidative stress,inflammation,apoptotis and the activation of the Nrf2 pathway[J].Int J Mol Med,2016,37(4):1075.
[5]Ruzicka J,Machova-Urdzikova L,Gillick J,et al.A comparative study of three different types of stem cells for treatment of rat spinal cord injury[J].Cell Transplant,2016,26(4):585-603.
[6]Grinspan J B.Bone morphogenetic proteins:inhibitors of myelination in development and disease[J].Vitam Horm,2015,99:195-222.
[7]Ding P,Yang Z,Wang W,et al.Transplantation of bone marrow stromal cells enhances infiltration and survival of CNP and Schwann cells to promote axonal sprouting following complete transection of spinal cord in adult rats[J].Am JTransl Res,2014,6(3):224-235.
[8]Louie P K,Hassanzadeh H,Singh K.Epidemiologic trends in the utilization,demographics,and cost of bone morphogenetic protein in spinal fusions[J].Curr Rev Musculoskelet Med,2014,7(3):177-181.
[9]Rodríguezevora M,Reyes R,Alvarezlorenzo C,et al.Bone regeneration induced by an in situ gel-forming poloxamine,bone morphogenetic protein-2 system[J].JBiomed Nanotechnol,2014,10(6):959.
[10]Van E L,Alt V.Bone graft substitutes and bone morphogenetic proteins for osteoporotic fractures:what is the evidence[J].Injury,2016,47:S43-S46.
[11]Wang X,Xu JM,Wang YP,et al.Protective effects of BMP-7 against tumor necrosis factorα-induced oligodendrocyte apoptosis[J].Int J Dev Neurosci,2016,53:10-17.
[12]胡岚翔.BMP-7对大鼠(环扎法)急性脊髓损伤后GFAP的表达的影响[D].皖南医学院,2014.
[13]Peng R J,Bing J,Ding X P,et al.Effect of TNF-αInhibition on Bone Marrow-Derived Mesenchymal Stem Cells in Neurological Function Recovery after Spinal Cord Injury via the Wnt Signaling Pathway in a Rat Model[J].Cell Physiol Biochem,2017,42(2):743-752.
[14]Lam S K,Sayama C,Harris D A,et al.Nationwide practice patterns in the use of recombinant human bone morphogenetic protein-2 in pediatric spine surgery as a function of patient-,hospital-,and procedure-related factors[J].J Neurosurg Pediatr,2014,14(5):476-485.
[15]Tannoury C A,An H S.Complications with the use of bone morphogenetic protein 2(BMP-2)in spine surgery.[J].Spine J,2014,14(3):552.