Repeated transient ischemia inhibits denervated gastrocnemius atrophy in rat models of sciatic nerve injury

doi:10.3969/j.issn.2095-4344.1311

Abstract

Abstract:

BACKGROUND: Preliminary study has shown that repeated transient ischemia may induce muscle ischemia tolerance to enrich target organs.
OBJECTIVE: To investigate the inhibitory effect of repeated transient ischemia on atrophy of denervated gastrocnemius muscle cells in a rat model of sciatic nerve injury.
METHODS: Sixty Sprague-Dawley rats aged 8 weeks were provided by Beijing Weitong Lihua Animal Technology Co., Ltd., and the study was approved by the Laboratory Animal Ethics Committee of Capital Medical University, approval number: AEEI-2017-055. The rats were randomly divided into control group (sciatic nerve injury model), experimental group (sciatic nerve injury model plus 3x(10-minute ischemia/10-minute reperfusion) and sham group [3x(10-minute ischemia/10-minute reperfusion)]. The wet mass maintenance rate of gastrocnemius, hematoxylin-eosin staining of gastrocnemius, electrophysiology of sciatic nerve, and content of vascular endothelial growth factor in gastrocnemius detected by ELISA were compared among groups at 2 and 4 weeks.
RESULTS AND CONCLUSION: (1) Electrophysiology showed that neither experimental nor control groups presented action potential or latency at 4 weeks. (2) The wet mass maintenance rate of gastrocnemius in the experimental group was significantly increased compared with the control group (P < 0.05), and the sham group was significantly superior to the experimental and control groups (P < 0.01). (3) The cross-sectional area of gastrocnemius fibers in the experimental group was significantly decreased compared with the control group (P < 0.05). (4) The content of vascular endothelial growth factor in gastrocnemius detected by ELISA in the experimental and sham groups was significantly higher than that in the control group (P < 0.05). (5) These results suggest that within 4 weeks of nerve injury, the repeated transient ischemia can effectively improve the expression of endogenous vascular endothelial growth factor in gastrocnemius, thus inhibiting its atrophy.

Key words: repeated transient ischemia, gastrocnemius, vascular endothelial growth factor, electrophysiology, muscle atrophy, sciatic nerve injury, rats

CLC Number:

R446
R318

Zhou Xiaobin1, Wang Dong2, Lin Pengzhao1, Gao Weijing1, Ji Yanlin1, Qi Huan3, Li Jing4, Zhou Junlin2 . Repeated transient ischemia inhibits denervated gastrocnemius atrophy in rat models of sciatic nerve injury [J]. Chinese Journal of Tissue Engineering Research, 2019, 23(23): 3667-3672.

Figures/Tables 2

2.1 实验动物数量分析实验选用大鼠60只，分为3组，实验过程无脱失，全部进入结果分析。 2.2 大鼠一般情况术后各组大鼠的伤口愈合良好，无感染及动物死亡。实验组及对照组大鼠在术后右后肢均跛行，表现为右侧踝关节丧失功能，踝下垂，趾屈曲。行走时表现为膝关节以下触地，且出现不同程度的足部溃疡。术后第3天对照组3只大鼠出现右足部溃疡，实验组4只大鼠右足跟部出现溃疡；实验组术后第3周大鼠足跟处溃疡完全愈合，术后第5周时对照组大鼠足部溃疡处完全愈合。术后实验组及对照组大鼠均出现右侧肢体完全瘫痪，右足足趾完全并拢，跖下垂、趾屈曲，行走时右下肢呈拖地行走状。术后4周实验组腓肠肌明显饱满，有弹性、色泽好；对照组相对实验组恢复较差，腓肠肌较瘦小，肌纤维萎缩变细、色泽苍白。假手术组因为损伤神经，行走活动及腓肠肌无明显改变。约于8周时2组停止恢复。 2.3 电生理结果术后4周各组肌电图结果(见图3)显示，于术后4周对患侧坐骨神经进行潜伏期测定，假手术组神经传导速度为(45.0±3.2)m/s，实验组及对照组均未测到神经的潜伏期，说明神经尚未到达靶器官形成回路，因此该结果并未进行统计学分析，实验组与对照组无差异。 2.4 腓肠肌湿质量术后2，4周时，大体观察可见，各组患侧腓肠肌均明显萎缩，色苍白，弹性差，对照组较重，肌肉萎缩明显。称质量结果示腓肠肌对照组较实验组肌肉质量明显增加，统计分析显示腓肠肌湿质量维持率在术后2周及4周时实验组较对照组均明显升高(P < 0.05)，见图4。 2.5 腓肠肌苏木精-伊红染色结果光镜下观察可见，随着时间延长，腓肠肌细胞纤维化程度逐渐减轻，2周及4周时实验组及对照组肌细胞的直径及截面积均较假手术组明显下降，对照组腓肠肌细胞萎缩严重，排列稀疏、不规则，周围纤维结缔组织增生明显，可见散在的炎性细胞浸润，而实验组腓肠肌细胞萎缩较轻，细胞质相对较饱满，肌细胞结构排列规则、紧密、完整，周围可见少量的纤维结缔组织增生。单位视野下平均肌纤维横截面积差异有显著性意义(P < 0.05)，见图5。"

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