嗅鞘细胞移植联合跑台训练抑制脊髓损伤大鼠慢性期瘢痕形成的机制  

Mechanism of olfactory ensheathing cells transplantation combined with treadmill training in inhibiting chronic scar formation in chronic spinal cord injury rats

作　　者：高月 王立童[1] 田亚茹 杜云霞 姜永梅[1] GAO Yue;WANG Litong;TIAN Yaru;DU Yunxia;JIANG Yongmei(Department of Neurological Rehabilitation,the Second Hospital of Dalian Medical University,Dalian 116027,China)

机构地区：[1]大连医科大学附属第二医院神经康复科,辽宁大连116027

出　　处：《中华实用诊断与治疗杂志》2018年第11期1051-1055,共5页Journal of Chinese Practical Diagnosis and Therapy

基　　金：辽宁省自然科学基金(2015020293)

摘　　要：目的探讨嗅鞘细胞移植联合跑台训练对脊髓损伤大鼠慢性期瘢痕形成的影响及作用机制。方法 40只成年SD大鼠随机分为假手术组、对照组、跑台组、细胞组、联合组各8只。对照组、跑台组、细胞组、联合组采用T10脊髓全横断法制备脊髓损伤模型,假手术组仅行椎板切除术、不损伤脊髓。术后第28天,跑台组给予跑台训练;细胞组硬脊膜注射嗅鞘细胞悬液;联合组于第1次跑台训练后硬脊膜注射嗅鞘细胞悬液;假手术组、对照组不进行任何干预。分别于术后第1、7、28、35、56天行BBB(Basso-Beasttie and Bresnahan,BBB)评分评估大鼠运动功能;术后第28、56天,各组分别处死2、6只大鼠,取脊髓组织行HE染色观察损伤局部组织空洞大小及瘢痕形成情况;术后第56天行免疫组织化学染色检测瘢痕区及损伤近端、远端神经丝蛋白200(neurofilament protein 200,NF200)表达,采用Western blot法检测NG2蛋白聚糖、神经型钙黏蛋白和β-连环蛋白表达。结果术后第35、56天,BBB评分在假手术组(21.00±0.63、21.00±0.89)、联合组(13.80±0.98、18.80±1.17)、跑台组(12.60±1.02、16.80±1.47)、细胞组(9.80±0.75、12.00±0.63)、对照组(8.00±0.89、8.20±0.75)逐渐降低(P<0.05);术后第28天,假手术组大鼠脊髓组织未出现损伤,对照组、细胞组、跑台组、联合组大鼠脊髓组织损伤区出现空洞及瘢痕,术后第56天损伤均加重,但细胞组、跑台组及联合组损伤程度较对照组轻,其中联合组损伤最轻;术后第56天,脊髓损伤瘢痕区及损伤近端NF200表达在假手术组(195.64±5.19、194.42±4.55)、联合组(163.51±6.24、184.85±4.24)、跑台组(144.51±5.25、161.18±7.36)、细胞组(126.25±4.99、142.92±4.99)和对照组(104.01±5.35、117.01±4.42)逐渐下降(P<0.05);5组损伤远端NF200表达比较差异无统计学意义(P>0.05);术后第56天神经型钙黏蛋白和β-连环蛋白表达在联合组(0.28±0.01、0.25±0.01)、跑台Objective To investigate the influence of olfactory ensheathing cells transplantation combined with treadmill training on inhibiting chronic scar formation and its mechanism in chronic spinal cord injury rats.Methods Forty rats were randomly divided into sham operation group,control group,treadmill training group,cell group and combination group,with 8 rats in each group.Complete spinal cord transection at T10 was used to establish spinal cord injury rats models in control group,treadmill training group,cell group and combination group.Sham operation group only received laminectomy without injuring spinal cord.Treadmill training group received training,cell group was injected with olfactory ensheathing cells suspension,combination group was injected with olfactory ensheathing cells suspension after initial treadmill training,and control group and sham operation group received no treatment 28 days after operation.The recovery of locomotor function was measured by Basso-Beattie and Bresnahan（BBB）scoring on the 1 st,7 th,28 th,35 th and 56 th postoperative days.Two rats were sacrificed on the 28 th postoperative day in each group,and the left 6 rats were sacrificed on the 56 th postoperative day.HE staining was applied to observe the cavity size and scar formation in the injured region.On the 56 th postoperative day,immunohistochemical staining was applied to detect the expressions of neurofilament protein 200（NF200）in scar areas,and proximal and distal injury,and Western blot was performed to detect NG2 proteoglycan,N-cadherin andβ-catenin.Results On 35 th and 56 th postoperative days,the BBB scores decreased gradually in sham operation group（21.00±0.63,21.00±0.89）,combination group（13.80±0.98,18.80±1.17）,treadmill training group（12.60±1.02,16.80±1.47）,cell group（9.80±0.75,12.00±0.63）and control group（8.00±0.89,8.20±0.75）（P<0.05）.On the 28 th postoperative day,the spinal tissue was not injured in sham operation group,the cavity and scar occurred in the control group,cell g

关 键 词：慢性脊髓损伤 跑台训练 嗅鞘细胞 瘢痕 神经突起 大鼠 

分 类 号：R651.2[医药卫生—外科学]