一种脊髓型颈椎病慢性压迫性动物模型的建立及评价

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摘要目的建立脊髓型颈椎病慢性压迫大鼠模型，并探讨不同压迫程度脊髓病变与椎间盘内炎症因子表达差异性的关系。方法选取三组雄性健康大鼠随机分为对照组(n＝10)、轻度压迫模型组(n＝30)、重度压迫模型组(n＝30)。于大鼠C_5~7节段椎板下方，分别植入两种尺寸的水凝胶压迫材料，建立轻度、重度压迫两组模型；对照组不置入水凝胶。在术后第4、8和12周随机选取6只大鼠进行斜板爬坡试验，处死后取C_5~7节段椎间盘和脊髓部分，采用HE染色对受压脊髓进行组织形态学观察；脱氧核糖核苷酸末端转移酶介导的缺口末端标记法(terminal dexynucleotidyltransferasedutp nick end labeling，TUNEL)对脊髓凋亡情况进行测定；然后采用聚合酶链式反应(polymerase chain reaction，PCR)和western blot法测定C_5~7椎间盘内肿瘤坏死因子(tumour necrosis factor，TNF)-α、白细胞介素(interleukin，IL)-1β、IL-33和IL-6的mRNA和蛋白的相对表达量。结果斜板爬坡试验，对照组术前与术后运动功能差异无统计学意义(P＞0.05)，但轻度、重度压迫模型组术后8周和12周大鼠所能承受的斜板角度均有下降，各组间差异有统计学意义(P＜0.05)。脊髓标本HE染色发现，术后第4周轻度、重度压迫模型组与术前相比，脊髓病变差异无统计学意义(P＞0.05)。术后第8周和12周受压脊髓出现神经元细胞变性、皱缩、脱髓鞘及空泡变性，随着压迫时间延长和程度增加而加重，差异有统计学意义(P＜0.05)。TUNEL法测的轻度、重度压迫模型组中的阳性细胞数量均显著高于对照组(P＜0.05)。术后8周和12周轻度、重度压迫模型组小鼠椎间盘中的IL-1β等炎性因子的mRNA和蛋白含量均呈高表达水平，且表达水平随压迫程度和造模时间而上调。结论实验结果表明，采用聚乙烯醇-聚丙烯酰胺互穿网络水凝胶为压迫材料可成功建立脊髓型颈椎病慢性压迫性大鼠模型。模型大鼠的脊髓损伤程度随压迫时间延长和程度增加而加重，轻度、重度压迫模型组椎间盘组织中各炎症因子均呈高表达，且随着退变的程度和压迫时间的延长表达水平显著升高。

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	李成立 1
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关键词 ：脊髓型颈椎病, 聚乙烯醇-聚丙烯酰胺互穿网络水凝胶, 慢性压迫

Abstract：Objective To establish a rat model of chronic compressive cervical spondylotic myelopathy (CSM) and to explore the relationship between different compression degree of myelopathy and the expression of inflammatory factors in intervertebral disc.Methods 3 group of fitness male rats were randomly assigned as matched group (n＝10)，mild compression model group (n＝30) and severe compression model group (n＝30).In the model group，the interlaminar ligaments were excised under the C_5~7 lamina，and carefully placed into two different size of water absorbent materials，hydrogel.and 6 rats were randomly chosen at 4，8 and 12 weeks Post-op，which were subjected to the inclined plate climbing test.After they were euthanized，the C_5~7segment of the intervertebral disc and spinal cord were taken.HE sstaining was used to observe the histomorphology of the compressed spinal cord.TUNEL staining was used to test the apoptotic cells.Then the PCR group and the Western blot way were used to test the content of protein and mRNA of TNF-α，IL-1β，IL-33 and IL-6 in C_5~7.Results There was no significant difference in motor function between the control group before and after operation (P＞0.05)，However，the angle of inclined plate of mild and severe compression compression model group decreased 8 and 12 weeks after operation，and the difference was obvious among each group (P＜0.05).HE staining showed that there was no significant difference between the compression group and the control group at 4 weeks after operation (P＞0.05).At 8 and 12 weeks after operation，the compressed spinal cord showed degeneration，shrinkage，demyelination and vacuolar degeneration；With the extension of the compression time and the increase of the degree of aggravation，there was statistical significance (P＜0.05).The number of positive cells detected by TUNEL in mild and severe compression compression model group was significantly higher than that in the corresponding control group (P＜0.05).At 8 and 12 weeks after operation，the mRNA and protein content of IL-1βand other factors in the intervertebral disc of the mild and severe compression model group were high，and the expression levels were up-regulated with the degree of compression and modeling time.Conclusion The results showed that polyvinyl alcohol polyacrylamide interpenetrating polymer network hydrogel was used as compression material to establish a chronic spinal cord compression model of cervical spondylotic myelopathy in rats.The injure of the spinal cord increased with the extension of compression time and degree，and the expression of inflammatory factors in intervertebral disc tissue of mild and severe compression model group was high，and the expression level increased significantly with the degree of degeneration and the extension of compression time.

Key words： cervical spondylotic myelopathy polyvinyl alcohol hydrogel chronic compression

基金资助:省自然科学基金项目(ZR2018LH003)

通讯作者: 张为

作者简介: 李成立(1984－ )，男，主治医师，滨州医学院烟台附属医院骨科，264100。

引用本文:

李成立 1，2，曲军杰 1，陈松 2，张为 2*. 一种脊髓型颈椎病慢性压迫性动物模型的建立及评价[J]. 实用骨科杂志, 2021, 27(12): 1096-1101.
Li Chengli 1，2，Qu Junjie 1，Chen Song 2，et al. Establishment and Evaluation of Animal Model of Chronic Compressive Cervical Spondylotic Myelopathy. sygkzz, 2021, 27(12): 1096-1101.

链接本文:

http://www.sygkzz.com/CN/ 或 http://www.sygkzz.com/CN/Y2021/V27/I12/1096

［1］张剑刚，郑彦华，张宪哲，等.影响椎管扩大成形术治疗多节段脊髓型颈椎病疗效的术前因素分析［J］.实用骨科杂志，2019，25(6)：542-546.
［2］Yang YP，Wang Y，Cao JM，et al.Risk factors for C5 palsy after anterior cervical decompression［J］.Int J Clin Exp Med，2018，11(7)：7125-7131.
［3］梁冬牧，韩来春，郝晨，等.脊髓线标准下Ⅱ型多节段脊髓型颈椎病手术选择及疗效研究［J］.实用骨科杂志，2021，27(4)：294-299.
［4］杜盛超，张玮，胡浩然，等.一种改良大鼠慢性脊髓压迫模型的建立与评价［J］.现代生物医学进展，2017，17(10)：1806-1809.
［5］Klironomos G，Karadimas S，Mavrakis A，et al.New experimental rabbit animal model for cervical spondylotic myelopathy［J］.Spinal Cord，2015，49(11)：1097-1102.
［6］Bergman RL，Levine JM，Coates JR，et al.Cervical spinal locking plate in combination with cortical ring allograft for a one level fusion in dogs with cervical spondylotic myelopathy［J］.Veterinary Surgery，2010，37(6)：530-536.
［7］DamHieu P，Magro E，Seizeur R，et al.Cervical cord compression due to delayed scarring around epidural electrodes used in spinal cord stimulation［J］.J Neurosurg Spine，2010，12(4)：409-412.
［8］陈盛，刘建航，刘本涛，等.脊髓型颈椎病动物模型制作的关注热点［J］.中国组织工程研究，2020，24(24)：3890-3896.
［9］Kim P，Haisa T，Kawamoto T，et al.Delayed myelopathy induced by chronic compression in the rat spinal cord［J］.Ann Neurol，2010，55(4)：503-511.
［10］Wang J，Rong W，Hu X，et al.Hyaluronantetrasaccharide in the cerebrospinal fluid is associated with self-repair of rats after chronic spinal cord compression［J］.Neuroscience，2012(210)：467-480.
［11］王军，韦峰，汪辉亮，等.慢性颈脊髓压迫模型大鼠构建及评价［J］.中国组织工程研究，2013，17(7)：1196-1200.
［12］Morishita Y，Falakassa J，Naito M，et al.The kinematic relationships of the upper cervical spine［J］.Spine，2009，34(24)：2642-2645.
［13］戎利民，蔡道章，吴金浪，等.慢性压迫性颈脊髓损害的超微结构观察［J］.中国临床解剖学杂志，2008，26(5)：536-538.
［14］Liu C，Zhang JF，Sun ZY，et al.Bioinformatics analysis of the gene expression profiles in human intervertebral disc degeneration associated with inflammatory cytokines［J］.J NeurosurgSci，2018，62(1)：16-23.
［15］何川，庞启雄，陈洪卫，等.腰椎间盘突出症中医证型与血清炎性因子相关性分析［J］.长江大学学报(自然科学版)，2020，17(4)：97-100.