Effect of tetramethylpyrazine on iron metabolism after spinal cord injury in rats

doi:10.12307/2023.360

Abstract

Abstract: BACKGROUND: Iron metabolism disorder is an important pathological factor leading to ferroptosis of nerve cells after spinal cord injury, which is not conducive to the repair of spinal cord injury. Tetramethylpyrazine, as the active ingredient monomer of Ligusticum Chuanxiong, a traditional Chinese medicine for activating qi and activating blood, has been proven to have good anti-inflammatory, anti-lipid peroxidation and neuroprotective effects on spinal cord injury. It is necessary to further clarify its mechanism of promoting spinal cord injury repair.
OBJECTIVE: To study the regulation mechanism of tetramethylpyrazine on iron metabolism after spinal cord injury in rats, and to explore its related mechanism of improving spinal cord injury.
METHODS: Totally 36 Sprague-Dawley rats were randomly divided into sham operation group, model group and tetramethylpyrazine group, with 12 rats in each group. In the sham operation group, only laminectomy was performed and normal saline was injected intraperitoneally after operation. Spinal cord injury models were prepared in model group and tetramethylpyrazine group. Normal saline and tetramethylpyrazine were injected intraperitoneally after operation, and the tissues were taken 4 weeks after operation. BBB limb motor function score was used to evaluate the limb motor function of rats. Nissl staining was used to observe the morphology of neurons. Prussian staining was used to observe the iron deposition in spinal cord tissue. Iron detection kit was used to detect the iron content in spinal cord tissue. Immunohistochemistry was used to detect the expression of ferritin heavy chain 1 and ferritin light chain protein. Western blot assay was used to detect the expression of ferritin heavy chain 1 and ferritin light chain protein.
RESULTS AND CONCLUSION: (1) The BBB score of model group and tetramethylpyrazine group was significantly lower than that of sham operation group at each time point (P < 0.01). Since the 14th day after operation, the BBB score of tetramethylpyrazine group was significantly higher than that of model group (P < 0.01). (2) Nissl staining results showed that the morphology and structure of neurons in the sham operation group were normal; a large amount of blood stasis and disordered morphology and structure of neurons were observed in the model group; blood stasis was less and morphology and structure of neurons were improved in the tetramethylpyrazine group. (3) Prussian staining results showed that there was less iron deposition in the sham operation group, and more iron deposition in the model group and tetramethylpyrazine group. The average optical density of positive Prussian staining in the model group and tetramethylpyrazine group was significantly higher than that in the sham operation group (P < 0.01). The average optical density of positive Prussian staining in tetramethylpyrazine group was significantly lower than that in the model group (P < 0.01). (4) The detection of iron content showed that the iron content in model group and tetramethylpyrazine group was significantly higher than that in sham operation group (P < 0.01). The iron content in tetramethylpyrazine group was significantly lower than that in model group (P < 0.01). (5) Immunohistochemical results showed that the average optical density of positive ferritin heavy chain 1 and ferritin light chain protein expression in model group and tetramethylpyrazine group was significantly lower than that in sham operation group (P < 0.01). The average optical density of positive ferritin heavy chain 1 and ferritin light chain protein expression in tetramethylpyrazine group was significantly higher than that in model group (P < 0.01). (6) Western blot assay showed that the relative expression of ferritin heavy chain 1 protein and ferritin light chain protein in model group and tetramethylpyrazine group was significantly lower than that in sham operation group (P < 0.01). The relative expression of ferritin heavy chain 1 protein and ferritin light chain protein in tetramethylpyrazine group was significantly higher than that in model group (P < 0.01). (7) It is concluded that tetramethylpyrazine can regulate the disorder of iron metabolism after spinal cord injury by regulating the expression of ferritin heavy chain 1 and ferritin light chain protein, so as to play a neuroprotective role and promote the recovery of limb motor function in spinal cord injury rats.

Key words: spinal cord injury, tetramethylpyrazine, iron metabolism, ferroptosis, ferritin heavy chain 1, ferritin light chain protein

CLC Number:

Fan Xiao, Tao Jingwei, Jiang Shengyuan, Deng Bowen, Mu Xiaohong. Effect of tetramethylpyrazine on iron metabolism after spinal cord injury in rats[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(22): 3561-3566.

Figures/Tables 8

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