目的 基于 SIRT1/AMPK 通路探讨虎杖苷对臂丛神经根性撕脱伤（BPRA）大鼠神经再生与功能恢复的作 用及机制。方法 将 60 只 SD 大鼠随机分为假手术组、模型组、虎杖苷低剂量组、虎杖苷高剂量组及 SIRT1 抑制剂组，每组 12 只。采用手术撕脱 C5~C7 脊神经根后再植 C6 神经根复制 BPRA 大鼠模型。BPRA 术后， 虎杖苷低、高剂量组腹腔注射虎杖苷 20、40 mg·kg-1 ，SIRT1 抑制剂组腹腔注射虎杖苷 40 mg·kg-1 +SIRT1 抑制 剂（EX527）5 mg·kg-1 ，每日 1 次，连续 8 周。采用 Terzis 梳洗试验（TGT）评估大鼠运动功能恢复情况；荧光金 逆行标记脊髓组织中的运动神经元；免疫荧光法检测脊髓组织中胆碱乙酰转移酶（ChAT）的表达水平；HE 染色 法观察肱二头肌的病理变化；Western Blot 法检测脊髓组织中 SIRT1、AMPK、LC3、p62 蛋白表达水平。 结果 各组大鼠体质量的差异无统计学意义（P＞0.05）。与假手术组比较，模型组大鼠的 TGT 评分显著降低 （P＜0.01）；肱二头肌肌纤维直径更小，成纤维细胞数量更多，肌肉萎缩明显，肱二头肌损伤侧/健侧湿质量比 显著下降（P＜0.01）；脊髓前角荧光金标记的运动神经元数量及脊髓前角 ChAT 阳性神经元数量明显减少（P＜ 0.05，P＜0.01）；脊髓组织中 p62 蛋白表达显著上调（P＜0.01），LC3Ⅱ/LC3Ⅰ、SIRT1、AMPK 蛋白表达均显 著下调（P＜0.05）。与模型组比较，虎杖苷高剂量组大鼠的 TGT 评分在术后第 3 周开始显著升高（P＜0.01），虎 杖苷低剂量组大鼠的 TGT 评分在术后第 6 周开始显著升高（P＜0.01）；虎杖苷低、高剂量组大鼠的肱二头肌肌 纤维面积明显增加，肌细胞核清晰，肌纤维形态更接近假手术组，肱二头肌损伤侧/健侧湿质量比显著升高 （P＜0.01），荧光金标记的运动神经元数量及脊髓前角 ChAT 阳性神经元数量显著增加（P＜0.01）；虎杖苷低、 高剂量组大鼠脊髓组织中 p62 蛋白表达显著下调（P＜0.01），AMPK 蛋白表达明显上调（P＜0.05）；虎杖苷高剂 量组大鼠脊髓组织中的 LC3Ⅱ/LC3Ⅰ、SIRT1 蛋白表达明显上调（P＜0.05，P＜0.01）。与虎杖苷高剂量组比较， SIRT1 抑制剂组大鼠的 TGT 评分显著降低（P＜0.01），肱二头肌损伤侧/健侧湿质量比显著降低（P＜0.01），荧 光金标记的运动神经元数量及脊髓前角 ChAT 阳性神经元数量显著减少（P＜0.01）；脊髓组织中的 LC3Ⅱ/LC3Ⅰ、 SIRT1、AMPK 蛋白表达显著下调（P＜0.01）。结论 虎杖苷能够促进 BPRA 大鼠运动功能恢复，改善肌肉萎 缩情况，增强运动神经元存活和轴突再生，其作用机制可能与激活 SIRT1/AMPK 通路促进自噬来抑制运动神 经元死亡有关。

Objective To explore the effect and mechanism of polydatin on nerve regeneration and functional recovery in rats with brachial plexus root avulsion （BPRA） injury based on the SIRT1/AMPK pathway. Methods Sixty SD rats were randomly divided into five groups：sham-operation group，model group，low-dose polydatin group，high-dose polydatin group， and SIRT1 inhibitor group， with 12 rats in each group. The BPRA rat model was established by surgically avulsing the C5-C7 spinal nerve roots and replanting the C6 nerve root. After BPRA surgery，the low- and high-dose polydatin groups were intraperitoneally injected with polydatin at doses of 20 and 40 mg·kg-1 ，respectively， while the SIRT1 inhibitor group was intraperitoneally injected with polydatin 40 mg·kg-1 + SIRT1 inhibitor （EX527） 5 mg·kg-1 ，once daily for 8 consecutive weeks. The Terzis grooming test （TGT） was used to evaluate the recovery of motor function in rats. Retrograde labeling of motor neurons in the spinal cord tissue was performed using Fluoro-Gold. The expression level of choline acetyltransferase （ChAT） in the spinal cord tissue was detected by immunofluorescence. Pathological changes in the biceps brachii muscle were observed using HE staining. The protein expression levels of SIRT1， AMPK， LC3， and p62 in the spinal cord tissue were detected by Western Blot. Results There was no significant difference in body mass among the groups （P＞0.05）. Compared with the sham-operation group，the TGT score of the model group was significantly decreased （P＜0.01）. The biceps brachii muscle fibers in the model group were smaller in diameter，with more fibroblasts and significant muscle atrophy，and the wet mass ratio of the injured/ healthy side of the biceps brachii was significantly decreased （P＜0.01）. The number of Fluoro-Gold-labeled motor neurons and ChAT-positive neurons in the anterior horn of the spinal cord was significantly reduced （P＜0.05，P＜ 0.01）. The protein expression of p62 in the spinal cord tissue was significantly up-regulated （P＜0.01）， while the protein expressions of LC3Ⅱ/LC3Ⅰ，SIRT1，and AMPK were significantly down-regulated （P＜0.05）. Compared with the model group，the TGT score of the high-dose polydatin group began to increase significantly from the third week after surgery （P＜0.01），and the TGT score of the low-dose polydatin group began to increase significantly from the sixth week after surgery （P＜0.01）. In the low- and high-dose polydatin groups，the cross-sectional area of the biceps brachii muscle fibers was significantly increased， the muscle cell nuclei were clear， and the muscle fiber morphology was closer to that of the sham-operation group. The wet mass ratio of the injured/healthy side of the biceps brachii was significantly increased （P＜0.01）， and the number of Fluoro-Gold-labeled motor neurons and ChATpositive neurons in the anterior horn of the spinal cord was significantly increased （P＜0.01）. The protein expression of p62 in the spinal cord tissue was significantly down-regulated （P＜0.01），while the protein expression of AMPK was significantly up-regulated （P＜0.05）. In the high-dose polydatin group，the protein expressions of LC3Ⅱ/LC3Ⅰ and SIRT1 in the spinal cord tissue were significantly up-regulated （P＜0.05， P＜0.01）. Compared with the high-dose polydatin group，the TGT score of the SIRT1 inhibitor group was significantly decreased （P＜0.01），the wet mass ratio of the injured/healthy side of the biceps brachii was significantly decreased （P＜0.01），and the number of FluoroGold-labeled motor neurons and ChAT-positive neurons in the anterior horn of the spinal cord was significantly reduced （P＜0.01）. The protein expressions of LC3Ⅱ/LC3Ⅰ，SIRT1，and AMPK in the spinal cord tissue were significantly down-regulated （P＜0.01）. Conclusion Polydatin can promote the recovery of motor function， improve muscle atrophy，and enhance motor neuron survival and axonal regeneration in rats with BPRA. Its mechanism may be related to the activation of the SIRT1/AMPK pathway to promote autophagy and inhibit motor neuron death.

R285.5

广东省基础与应用基础研究基金项目（2021A1515110800）；广东省中医药局科研项目（20241084）。