Du Meridian electroacupuncture inhibits ferroptosis and promotes neurorepair in rats with acute cervical spinal cord injury

doi:10.12307/2025.767

Abstract

Abstract: BACKGROUND: Recent studies have shown that Du Meridian electroacupuncture has a unique effect on alleviating spinal cord injury, but the underlying mechanisms require further clarification.
OBJECTIVE: To investigate the regulatory effects and the associated action mechanisms of Du Meridian electroacupuncture on ferroptosis after cervical spinal cord injury in rats.
METHODS: One hundred SD rats were randomly divided into sham, model, Du Meridian electroacupuncture, RSL3, and Du Meridian electroacupuncture + RSL3 groups. The sham group underwent only laminectomy. The other four groups were subjected to cervical spinal cord injury by the Allen method. The Du Meridian electroacupuncture group received electroacupuncture after cervical spinal cord injury. The RSL3 group received intraperitoneal injections of glutathione peroxidase 4 inhibitor RSL3 after cervical spinal cord injury. The Du Meridian electroacupuncture + RSL3 group received both electroacupuncture and RSL3 intervention after cervical spinal cord injury. Samples were collected on postoperative days 7 and 28 to assess motor function, histological morphology, neuronal survival, glial scar formation, oxidative stress levels, Fe2+ content, glutathione peroxidase 4, and long-chain acyl-CoA synthetase 4 expression.
RESULTS AND CONCLUSION: (1) Finally, 90 rats completed the follow-up experiment, with 18 rats in each group. (2) FLS and BBB scores were significantly higher in the Du Meridian electroacupuncture group compared with the model and Du Meridian electroacupuncture + RSL3 groups (P < 0.05). (3) Compared with the model group, Du Meridian electroacupuncture improved cervical spinal cord tissue morphology and mitochondrial ultrastructure, while these effects were inhibited by RSL3. (4) Du Meridian electroacupuncture increased the expression of microtubule-associated protein 2, glutathione peroxidase 4, glutathione, and superoxide dismutase (P < 0.05) and reduced the expression of glial fibrillary acidic protein, long-chain acyl-CoA synthetase 4, reactive oxygen species, malondialdehyde, and Fe2+ compared with the model group (P < 0.05). However, RSL3 reversed the inhibitory effects of Du Meridian electroacupuncture on ferroptosis, lipid peroxidation and oxidative stress. (5) The results suggest that Du Meridian electroacupuncture inhibits ferroptosis by regulating the glutathione peroxidase 4/long-chain acyl-CoA synthetase 4 axis, thereby reducing secondary neuronal damage and glial scar formation after cervical spinal cord injury and improving neurological function.

Key words: cervical spinal cord injury, Du Meridian electroacupuncture, ferroptosis, glutathione peroxidase 4, long-chain acyl-CoA synthetase 4, oxidative stress, neurorepair, engineered tissue construction

CLC Number:

Sun Rongyan, Xu Luchun, Jiang Guozheng, Song Jiawei, Ma Yukun, Fan Jiaojiao, Wang Guanlong, Yang Yongdong, Yu Xing. Du Meridian electroacupuncture inhibits ferroptosis and promotes neurorepair in rats with acute cervical spinal cord injury [J]. Chinese Journal of Tissue Engineering Research, 2025, 29(29): 6228-6238.

Figures/Tables 6

2.1 实验动物数量分析 100只雌性SD大鼠随机分为5组，在颈髓损伤模型构建过程中死亡3只，最后每组随机选取18只，共计90只大鼠完成后续实验。 2.2 各组大鼠颈髓损伤后四肢运动功能采用FLS评分和BBB评分评估四肢运动功能。除假手术组外，其余各组在术后第1天均表现出严重的运动功能缺陷。从术后3 d开始，督脉电针组的FLS评分和BBB评分开始逐渐恢复，与模型组和督脉电针+RSL3组比较差异有显著性意义(P < 0.05)，见图1。结果表明督脉电针促进颈髓损伤后四肢功能恢复，其促进作用可被铁死亡诱导剂RSL3阻断。 2.3 各组大鼠颈髓损伤后组织病理学及线粒体微观形态苏木精-伊红染色显示，在术后28 d，假手术组大鼠脊髓组织结构正常完整，无水肿；模型组大鼠脊髓组织结构重度异常，水肿严重，大面积组织糜烂，组织间隙增大明显，免疫/炎性细胞广泛浸润；督脉电针组较模型组颈髓结构更为致密，水肿减轻，组织间隙较模型组缩小，免疫/炎性细胞浸润程度降低；督脉电针+RSL3组颈髓结构类似于模型组，存在较大面积组织糜烂与广泛免疫/炎性细胞浸润，见图2A。尼氏染色显示，在术后28 d，假手术组灰质内存在大量神经元，尼氏体形态规则且呈虎斑状；模型组脊髓周围组织大量溶解，广泛神经元死亡，未见存活神经元，可见散在的细胞碎片和空洞；督脉电针组可见部分存活神经元，溶解性空腔较模型组小；督脉电针+RSL3组颈髓结构类似于模型组，可见大量神经元死亡、细胞碎片和空洞，见图2B。线粒体形态学的改变是铁死亡的重要标志，因此采用透射电镜观察线粒体的微观形态。模型组线粒体体积变小，基质电子密度升高；督脉电针组线粒体形态明显改善，线粒体体积减少情况较模型组改善，基质电子密度降低，内膜结构趋于完整，嵴清晰可见，显示出一定的保护作用；督脉电针+RSL3组线粒体基质电子密度较督脉电针组升高，嵴结构部分更加模糊，表明RSL3逆转了督脉电针对线粒体的保护性，进一步支持GPX4在铁死亡中的关键作用，见图2C。组织学和线粒体的微观形态表明，督脉电针可有效减轻颈髓损伤后铁死亡。 2.4 各组大鼠颈髓损伤后神经元继发性损伤及胶质瘢痕形成情况采用微管相关蛋白2和胶质纤维酸性蛋白免疫荧"

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