目的 探讨五花血藤水煎液（Sargentodoxa cuneata，SCD）在小鼠脊髓损伤（spinal cord injury，SCI）后的抗炎作用及其对运动功能恢复的影响。 方法 选取36只SPF级小鼠随机分成假手术组、脊髓损伤组和SCD组，利用BMS评分检测小鼠后肢运动功能恢复情况，免疫蛋白质印迹法检测各组小鼠脊髓白细胞介素（IL）-6、IL-12A、肿瘤坏死因子（TNF）-α和脑源性神经营养因子（brain-derived neurotrophic factor，BDNF）的表达，免疫荧光染色通过NeuN观察损伤处脊髓前角神经元变化，rAAV-retro逆行示踪观察各组小鼠胫骨前肌（tibialis anterior muscle，TA）相关脊髓运动神经元分布。 结果 与脊髓损伤组相比，SCD组小鼠在损伤后7 d的BMS评分显著提高（P<0.05），损伤后14 d，IL-6、IL-12A和TNF-α表达明显降低（P<0.05），BDNF的表达显著升高（P<0.05）；损伤处脊髓前角神经元数量显著增多（P<0.05），TA相关运动神经元数量明显增多（P<0.05）。 结论 五花血藤水煎液能明显下调炎症因子IL-6、IL-12A和TNF-α的表达，发挥抗炎作用，同时上调BDNF的表达从而对运动神经元起保护作用，促进损伤后小鼠运动功能的恢复。
Abstract
Objective To investigate the anti-inflammatory effect of sargentodoxa cuneata decoction (SCD) after spinal cord injury (SCI) and its effect on the recovery of motor function. Methods Thirty-six SPF grade mice were randomly divided into a sham operation group, a SCI group and a SCD group. The recovery of hindlimb motor function was detected by BMS (Basso mouse scale), and the expression levels of interleukin (IL) -6, IL-12A, tumor necrosis factor (TNF) -α and brain-derived neurotrophic factor (BDNF) were detected by Western blot. Immunofluorescence staining of NeuN was performed to observe the changes of neurons in anterior horn of spinal cord. The distribution of spinal cord motoneurons related to tibialis anterior muscle (TA) was observed by rAAV-retro retrograde tracing. Results Compared with SCI group, the BMS score of SCD group was significantly higher at 7 days after injury (P<0.05), and IL-6, IL-12A and TNF-α were significantly higher at 14 days after injury, while the expression of BDNF was significantly higher than that in SCI group (P<0.05). Compared with SCI group, the number of spinal cord anterior horn neurons in SCD group was significantly increased (P<0.05), and the number of rAAV-retro retrograde labeled TA related motor neurons was also significantly increased (P<0.05). Conclusions The sargentodoxa cuneata decoction can significantly down-regulate the levels of inflammatory factors IL-6, IL-12A and TNF- α to play an anti-inflammatory role and up-regulate the expression of BDNF, so as to protect motor neurons and promote the recovery of motor function in mice after spinal cord injury.
关键词
脊髓损伤； /
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五花血藤； /
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抗炎作用； /
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功能恢复
Key words
Spinal cord injury; /
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Sargentodoxa cuneata; /
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Anti-inflammatory effect; /
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Functional recovery
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基金
贵州省中医药管理局中医药、民族医药科学技术研究课题（QZYY-2020-059）；贵州医科大学国家自然科学基金培育项目(19NSP032)；贵州省科技厅计划项目（黔科合基础-ZK[2021]一般472）
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