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中华临床医师杂志(电子版) ›› 2018, Vol. 12 ›› Issue (04) : 238 -244. doi: 10.3877/cma.j.issn.1674-0785.2018.04.010

所属专题: 文献

基础研究

神经生长因子和神经碎片套管对修复损伤神经的实验研究
丰波1,(), 王执宇2, 邹英财2, 代光明2, 徐立静3   
  1. 1. 014010 包头,内蒙古包钢医院手足踝外科
    2. 010010 呼和浩特,内蒙古医科大学研究生学院
    3. 014010 包头,内蒙古包钢医院泌尿外科
  • 收稿日期:2017-12-20 出版日期:2018-02-15
  • 通信作者: 丰波

Repair of nerve injury using biological conduit rich in nerve growth factor and nerve fragments

Bo Feng1,(), Zhiyu Wang2, Yingcai Zou2, Guangming Dai2, Lijing Xu3   

  1. 1. Department of Hand, Foot and Ankle Surgery, Inner Mongolia Baogang Hospital, Baotou 014010, China
    2. Graduate School of Inner Mongolia Medical University, Hohehot 010000, China
    3. Department of Urinary Surgery, Inner Mongolia Baogang Hospital, Baotou 014010, China
  • Received:2017-12-20 Published:2018-02-15
  • Corresponding author: Bo Feng
  • About author:
    Corresponding author: Feng Bo, Email:
引用本文:

丰波, 王执宇, 邹英财, 代光明, 徐立静. 神经生长因子和神经碎片套管对修复损伤神经的实验研究[J]. 中华临床医师杂志(电子版), 2018, 12(04): 238-244.

Bo Feng, Zhiyu Wang, Yingcai Zou, Guangming Dai, Lijing Xu. Repair of nerve injury using biological conduit rich in nerve growth factor and nerve fragments[J]. Chinese Journal of Clinicians(Electronic Edition), 2018, 12(04): 238-244.

目的

探讨富含神经生长因子(NGF)和神经碎片的生物套管在周围神经损伤再生修复过程中的作用。

方法

建立动物模型,将SD大鼠随机分成4组,分别为A对照组、B自体神经外膜小间隙吻合单纯添加自体神经碎片组、C自体神经外膜小间隙吻合同时添加NGF和自体神经碎片组、D生物套管吻合同时添加NGF和自体神经碎片组;观察各组一般形态、显微镜下吻合口处神经恢复形态以及组织学表现。采用方差分析比较各组有髓神经纤维数、施旺细胞数以及运动神经传导速度(MNCV)值的组间差异,组间的两两比较采用LSD-t检验。

结果

(1)A组大鼠在第8周时足底溃疡出现明显自愈,右下肢开始出现自主活动,部分足底溃疡的大鼠在第12周时仍未出现明显的自愈,右下肢无活动;其余实验组出现右下肢足底溃疡的大鼠在第8周时出现明显的自愈,并且右下肢表现出自主活动。第12周时A组大鼠右下肢足趾仍然呈挛缩状态,不能自由活动,而其余3组右下肢足趾大部分恢复自主活动。(2)纵切面见B组、C组、D组所构建的小间隙管腔内均可见到再生神经纤维。第12周时,A组有髓神经纤维数目为(22.30±4.66)根/400视野,B组有髓神经纤维数目为(51.60±4.45)根/400视野,C组有髓神经纤维数目为(56.20±4.66)根/400视野,D组有髓神经纤维数目为(59.20±5.81)根/400视野,组间差异具有统计学意义(F=298.48,P<0.001);抗S-100染色单位视野下,A组施旺细胞数目为(16.00±2.24)根/400视野,B组施旺细胞数目为(21.00±3.40)根/400视野,C组施旺细胞数目为(30.20±3.03)根/400视野,D组施旺细胞数目为(34.80±3.35)根/400视野,组间差异具有统计学意义(F=197.63,P<0.001)。术后第12周时,A组MNCV值为(7.57±1.79)m/s,B组MNCV值为(11.49±1.12)m/s,C组MNCV值为(13.86±2.03)m/s,D组MNCV值为(20.16±2.48)m/s,组间比较差异具有统计学意义(F=188.80,P<0.001)。D组对周围神经损伤的修复后的有髓神经纤维数目、施旺细胞数目以及MNCV均具有明显优势,与A组、B组、C组比较,差异均具有统计学意义(t=26.55、5.45、2.15,P<0.001、<0.001、=0.034;t=21.87、16.05、5.35,P均<0.001;t=23.19、15.97、11.60,P均<0.001)。

结论

本实验证实自体神经碎片与NGF的联合应用,明显提高周围神经再生微环境中神经损伤修复效果。

Objective

To investigate the role of biological conduit containing nerve growth factor (NGF) and nerve fragments in the regeneration and repair process of peripheral nerve injury, in order to provide a theoretical basis for clinical peripheral nerve repair.

Methods

A rat model of peripheral nerve injury was established. The experimental animals were randomly divided into four groups: control group (A), autogenous adventitial small gap anastomosis plus autologous nerve fragment group (B), autogenous adventitial small gap anastomosis plus NGF and autologous nerve fragment group (C), and biological conduit plus NGF and autogenous nerve fragment group (D). Gross morphology observation, microscopic observation at the anatomical site, and histological observation were conducted. Measurement data are expressed as mean±standard deviation, and analysis of variance was performed to compare myelinated nerve fibers, Schwann cell number, and motor nerve conduction velocity (MNCV) values among the three groups, followed by pairwise comparisons using the LSD-t test. P<0.05 was considered statistically significant.

Results

In group A, at the 8th week, the sole ulcer appeared to heal spontaneously, and the right lower limb started to show voluntary activity. The rats with plantar ulcer of the lower limb still did not show significant self-healing at the 12th week, and there was no limb activity. The lower limbs of rats in the other groups showed significant self-healing at the 6th week, with spontaneous activity in the right lower limb. At the 12th week, the toe of the right lower extremity of group A rats was still contracted and could not move freely, while most of those in the remaining three groups had activities. In the longitudinal sections, regenerated nerve fibers were observed in the small gap lumens constructed in groups B, C, and D. At 12 weeks after operation, the numbers of myelinated nerve fibers in groups A-D were 22.30±4.66 (per visual field at ×400 magnification), 51.60±4.45, 56.20±4.66, and 59.20±5.81, respectively, and the difference among the four groups was significant (F=298.48, P<0.001). The numbers of Schwann cells in groups A-D were 16.00±2.24 (per visual field at ×400 magnification), 21.00±3.40, 30.20±3.03, and 34.80±3.35, respectively, and the difference among the four groups was significant (F=197.63, P<0.001). At the 12th week after surgery, the MNCV values in groups A-D were 7.57±1.79 (m/s), 11.49±1.12, 13.86±2.03, and 20.16±2.48, respectively, and the difference among the four groups was significant (F=188.80, P<0.001). The number of myelinated nerve fibers, the number of Schwann cells, and MNCV were significantly higher in group D than in groups A-C (t=26.55, 5.45, 2.15, P<0.001, < 0.001, =0.034; t=21.87, 16.05, 5.35, P<0.001; t=23.19, 15.97, 11.60, P<0.001).

Conclusion

Combined use of autologous nerve fragments and NGF significantly improves the repair of nerve injury in the peripheral nerve regeneration microenvironment, and this finding provides a theoretical basis for clinical repair of peripheral nerve injury.

图1~4 各组术后12周SD大鼠的足底外观图1为A组大鼠术后12周足底外观,足底溃疡至术后12周时仍未完全自愈,可见足趾仍然呈挛缩状态,不能自由活动;图2为B组大鼠术后12周足底外观,足底溃疡至术后12周时完全自愈;图3为C组大鼠术后12周足底外观,图4为D组大鼠术后12周足底外观,C组和D组大鼠足底溃疡至术后12周时完全自愈,足趾大部分恢复自主活动
图5~8 各组大鼠标本HE染色显微镜结果(×100)图5为A组染色结果,可见神经断端吻合口形成明显瘤样增生;图6为B组染色结果,可见神经瘤样增生组织较A组小,神经干连续,出现管腔塌陷;图7为C组染色结果,神经瘤样增生组织也较A组小,神经干连续,管腔塌陷明显减少;图8为D组染色结果,神经瘤样增生组织也较A组小,神经干连续,管腔未明显塌陷
图9~12 各组大鼠标本12周时S-100染色结果(×400)图9为A组染色结果,吻合口处施旺细胞及神经丝蛋白数目较少,丝蛋白较少有完整的连续性,大部分呈撕裂或断裂;图10为B组染色结果,腔隙边缘处较少有施旺细胞及神经丝蛋白,并且边缘出现神经轴突的连续性中断;图11为C组染色结果,腔隙边缘处较少有施旺细胞及神经丝蛋白,并且边缘出现神经轴突的连续性中断;图12为D组染色结果,腔隙边缘可观察到一定数量的施旺细胞及神经丝蛋白,无连续性中断
表1 4组有髓神经纤维数目、施旺细胞数目以及MNCV比较(±s
[1]
盛华刚,朱立俏,田景振, 等. 神经生长因子对雪旺细胞增殖的影响 [J]. 中国新药杂志, 2013, 12: 1462-1464, 1476.
[2]
高延明,李靖年. 局部应用神经生长因子对坐骨神经损伤模型大鼠修复与再生的影响 [J]. 大连医科大学学报, 2013, 35(4): 336-340.
[3]
张培训,寇玉辉,韩娜, 等. 可降解生物套管小间隙套接法修复周围神经损伤的临床观察 [J]. 北京大学学报(医学版), 2012, 44(6): 842-846.
[4]
Tos P, Battiston B, Ciclamini D, et al. Primary repair of crush nerve injuries by means of biological tubulization with muscle-vein-combined grafts [J]. Microsurgery, 2012, 32(5): 358-563.
[5]
Wang Y, Zhao Z, Ren Z, et al. Recellularized nerve allografts with differentiated mesenchymal stem cells promote peripheral nerve regeneration [J]. Neurosci Lett, 2012, 514(1): 96-101.
[6]
Yuan Q, Su H, Guo J, et al. Decreased c-Jun expression correlates with impaired spinal motoneuron regeneration in aged mice following sciatic nerve crush [J]. Exp Gerontol, 2012, 47(4): 329-336.
[7]
Ruff CA, Staak N, Patodia S, et al. Neuronal c-Jun is required for successful axonal regeneration,but the effects of phosphorylation of its N-terminus are moderate [J]. Neurochem, 2012, 121(4): 607-618.
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