中国组织工程研究

中国组织工程研究 ›› 2025, Vol. 29 ›› Issue (32): 6843-6850.doi: 10.12307/2025.932

• 脊柱组织构建 spinal tissue construction • 上一篇    下一篇

二氢槲皮素对脊髓损伤大鼠炎症反应标志物表达的影响

徐  彪1，董玉珍1，2，路  坦1，3   

  1. 1新乡医学院第一附属医院，河南省卫辉市  453100；2河南省神经修复重点实验室，河南省卫辉市  453100；3新乡市骨关节退行性疾病研究重点实验室，河南省卫辉市  453100
  • 收稿日期:2024-08-10 接受日期:2024-10-22 出版日期:2025-11-18 发布日期:2025-04-25
  • 通讯作者: 董玉珍，博士，教授，新乡医学院第一附属医院，河南省卫辉市 453100；河南省神经修复重点实验室，河南省卫辉市 453100 通讯作者：路坦，博士，副教授，新乡医学院第一附属医院，河南省卫辉市 453100；新乡市骨关节退行性疾病研究重点实验室，河南省卫辉市 453100
  • 作者简介:徐彪，男，1992年生，河南省清丰县人，汉族，硕士，主治医师，主要从事神经损伤修复后再生及三维有限元在运动医学方面的研究。
  • 基金资助:
    河南省神经修复重点实验室开放课题(HNSIXF-2021-016，KFKTYB202122)，项目负责人：董玉珍

Effect of dihydroquercetin on the expression of inflammatory response markers in rats with spinal cord injury

Xu Biao1, Dong Yuzhen1, 2, Lu Tan1, 3   

  1. 1The First Affiliated Hospital of Xinxiang Medical University, Weihui 453100, Henan Province, China; 2Henan Provincial Key Laboratory of Nerve Repair, Weihui 453100, Henan Province, China; 3Xinxiang Key Laboratory of Bone and Joint Degenerative Disease Research, Weihui 453100, Henan Province, China
  • Received:2024-08-10 Accepted:2024-10-22 Online:2025-11-18 Published:2025-04-25
  • Contact: Dong Yuzhen, MD, Professor, The First Affiliated Hospital of Xinxiang Medical University, Weihui 453100, Henan Province, China; Henan Provincial Key Laboratory of Nerve Repair, Weihui 453100, Henan Province, China Co-corresponding author: Lu Tan, MD, Associate professor, The First Affiliated Hospital of Xinxiang Medical University, Weihui 453100, Henan Province, China; Xinxiang Key Laboratory of Bone and Joint Degenerative Disease Research, Weihui 453100, Henan Province, China
  • About author:Xu Biao, Master, Attending physician, The First Affiliated Hospital of Xinxiang Medical University, Weihui 453100, Henan Province, China
  • Supported by:
    Open Project of Henan Key Laboratory of Neural Repair, No. HNSIXF-2021-016 and KFKTYB202122 (both to DYZ)

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摘要：


文题释义：
脊髓损伤：其发病机制是一个复杂的过程，分为原发性损伤和继发性损伤。原发性损伤是指在损伤时外力物理因素的影响，通常是不可逆的；继发性损伤是原发性损伤的延续和发展，是一种复杂的级联放大反应，涉及多种机制，其中脊髓损伤后的“炎症风暴”是最重要的病理事件，而小胶质细胞是级联各种通道及引发再损伤的始作俑者。
二氢槲皮素：属于维生素P族，广泛存在于多种中草药，不仅具有多种生物活性还有广泛的药理学作用，包括抗氧化、抗炎和神经保护作用，而且其生物安全性极高。

背景：小胶质细胞诱导的“炎症风暴”是导致脊髓损伤后神经元细胞死亡的重要病理因素，极其不利于脊髓损伤后的恢复。二氢槲皮素属于维生素P族，有极好的生物活性，对脊髓损伤具有良好的抗炎、抗脂质过氧化反应以及神经保护作用，需进一步明确其促进脊髓损伤修复的作用机制。
目的：探究二氢槲皮素是否能够通过其抗炎、抗氧化性能改善脊髓损伤后诱发的炎症状态，达到保护脊髓损伤的目的。
方法：将48只SD大鼠按随机数字表法随机分为假手术组、脊髓损伤组和低剂量(30 mg/kg)二氢槲皮素组、高剂量(50 mg/kg)二氢槲皮素组。假手术组仅行椎板切除术，术后给予生理盐水腹腔注射；后3组制备脊髓损伤模型，术后分别给予生理盐水、30 mg/kg及50 mg/kg二氢槲皮素腹腔注射，术后4周取材。采用BBB肢体运动功能评分及机械缩足反应阈值评价大鼠肢体运动功能以及痛觉恢复情况，苏木精-伊红染色观察脊髓组织病理学变化，尼氏染色观察尼氏体数量及神经元形态，检测脊髓组织超氧化物歧化酶活性，免疫荧光检测活性氧表达及小胶质细胞Iba1的表达，Western blot检测脊髓中Iba1及白细胞介素1β、白细胞介素6、白细胞介素10、肿瘤坏死因子α、p38丝裂原活化蛋白激酶的蛋白表达情况。
结果与结论：①行为学测试：自术后14 d起，高剂量二氢槲皮素组大鼠BBB肢体运动功能评分显著高于脊髓损伤组(P < 0.01)；在术后21 d时，高剂量二氢槲皮素组的机械缩足反应阈值显著低于脊髓损伤组(P < 0.05)，表现出更出色的运动功能；②苏木精-伊红染色及尼氏染色：与脊髓损伤组相比，高剂量二氢槲皮素组脊髓组织可见神经元形态稍肿胀但结构完整较清晰，尼氏体数量增多；③超氧化物歧化酶检测：与脊髓损伤组相比，高剂量二氢槲皮素组大鼠脊髓组织超氧化物歧化酶活性显著升高 (P < 0.05)；④活性氧免疫荧光检测：与脊髓损伤组相比，高剂量二氢槲皮素的治疗能显著减轻脊髓损伤后活性氧的堆积(P < 0.05)；⑤小胶质细胞标志物Iba1的检测：与脊髓损伤组相比，高剂量二氢槲皮素组Iba1的蛋白表达水平显著降低(P < 0.01)；⑥Western blot检测：与脊髓损伤组相比，高剂量二氢槲皮素组的炎性因子(白细胞介素1β、白细胞介素6 和肿瘤坏死因子α)及磷酸化P38丝裂原活化蛋白激酶的表达显著减少(P < 0.01)，而抗炎因子白细胞介素10则显著增加(P < 0.01)；⑦提示二氢槲皮素能有效逆转大鼠脊髓损伤后诱发的多种炎症标志物的表达变化，从而发挥神经保护作用，改善运动功能。

https://orcid.org/0000-0002-3509-5025(徐彪)

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

关键词: 脊髓损伤, 炎症因子, 小胶质细胞, 尼氏体, 二氢槲皮素, 神经元, 活性氧, 抗炎因子

Abstract: BACKGROUND: The “inflammatory storm” induced by microglia is an important pathological factor causing the death of neuronal cells after spinal cord injury, which is extremely unfavorable for the recovery after spinal cord injury. Dihydroquercetin belongs to the vitamin P group and has excellent biological activity. It has good anti-inflammatory, anti-lipid peroxidation and neuroprotective effects on spinal cord injury. The mechanism by which it promotes the repair of spinal cord injury needs to be further clarified.
OBJECTIVE: To investigate whether dihydroquercetin can ameliorate the inflammatory state induced after spinal cord injury through its anti-inflammatory and antioxidant properties for the purpose of protection against spinal cord injury.
METHODS: Forty-eight Sprague-Dawley rats were divided into sham operation group, spinal cord injury group, low-dose dihydroquercetin group, and high-dose dihydroquercetin group according to the random number table method. The sham operation group only underwent laminectomy and was given intraperitoneal injection of normal saline after the operation. Animal models of spinal cord injury were made in the latter three groups. After the operation, normal saline, 30 mg/kg, and 50 mg/kg dihydroquercetin were given by intraperitoneal injection respectively, and samples were collected 4 weeks after the operation. The Basso-Beattie-Bresnahan (BBB) limb motor function score and mechanical paw withdrawal threshold were used to evaluate the limb motor function and pain recovery of rats. Hematoxylin-eosin staining was used to observe the pathological changes of spinal cord tissue; Nissl staining was used to observe the number of Nissl bodies and the morphology of neurons. The activity of superoxide dismutase in spinal cord tissue was detected. Immunofluorescence was used to detect the expression of reactive oxygen species and microglia Iba1. Western blot was used to detect the protein expression of Iba1 and interleukin-1β, interleukin-6, interleukin-10, tumor necrosis factor-α, and p38 mitogen-activated protein kinase in the spinal cord.
RESULTS AND CONCLUSION: (1) In the behavioral test, the BBB limb motor function score showed that since 14 days after the operation, the BBB limb motor function score of the high-dose dihydroquercetin group was significantly better than that of the spinal cord injury group (P < 0.01); the mechanical paw withdrawal threshold was significantly lower in the high-dose dihydroquercetin than the spinal cord injury group at 21 days after the operation (P < 0.05), and the former group showed better motor function. (2) Hematoxylin-eosin staining and Nissl staining showed that compared with the spinal cord injury group, the high-dose dihydroquercetin group showed slightly swollen but structurally intact and clearer neuron morphology in the spinal cord tissue, and the number of Nissl bodies increased. (3) Superoxide dismutase detection: Compared with the spinal cord injury group, the activity of superoxide dismutase in the spinal cord tissue of the high-dose dihydroquercetin increased significantly (P < 0.05). (4) Reactive oxygen species immunofluorescence detection: Compared with the spinal cord injury group, the high-dose dihydroquercetin could significantly reduce the accumulation of reactive oxygen species after spinal cord injury (P < 0.05). (5) Detection of microglial marker Iba1: Compared with the spinal cord injury group, the high-dose dihydroquercetin could significantly reduce the expression of microglial marker Iba1 after spinal cord injury (P < 0.01). (6) Western blot detection results showed that compared with the spinal cord injury group, the expressions of inflammatory factors (interleukin-1β, interleukin-6, and tumor necrosis factor-α) and phosphorylated p38 mitogen-activated protein kinase in the high-dose dihydroquercetin group decreased significantly (P < 0.01), while the expression of anti-inflammatory factor interleukin-10 increased significantly (P < 0.01). These findings indicate that dihydroquercetin can effectively reverse the expression changes of multiple inflammatory markers induced after spinal cord injury in rats, thereby exerting neuroprotective effects and improving motor function.

Key words: spinal cord injury, inflammatory factors, microglia, Nissl body, dihydroquercetin, neurons, reactive oxygen species, anti-inflammatory factors

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