Xiaozheng Zhitong Paste Alleviates Bone Cancer Pain of Mice by Reducing Ferroptosis in Spinal Cord Tissue and Neuronal Damage via Regulating Nrf2/HO-1/GPX4/SLC7A11 Signaling Pathway
10.13422/j.cnki.syfjx.20252038
- VernacularTitle:消癥止痛外用方调控Nrf2/HO-1/GPX4/SLC7A11信号通路减轻小鼠脊髓组织铁死亡和神经元损伤缓解骨癌痛
- Author:
Juanxia REN
1
;
Lu SHANG
1
;
Guangda ZHENG
2
;
Linghan MENG
2
;
Lingyun WANG
1
;
Changlin LI
1
;
Dongtao LI
2
;
Yaohua CHEN
3
;
Guiping YANG
3
;
Yanju BAO
2
Author Information
1. Liaoning University of Traditional Chinese Medicine， Shenyang 110847， China
2. Guang'anmen Hospital，China Academy of Chinese Medical Sciences，Beijing 100053，China
3. Xining Hospital of Traditional Chinese Medicine， Xining 810001， China
- Publication Type:Journal Article
- Keywords:
bone cancer pain （BCP）;
Xiaozheng Zhitong paste （XZP）;
ferroptosis;
spinal cord neuron;
nuclear factor erythroid 2-related factor 2 （Nrf2）/heme oxygenase 1 （HO-1）/glutathione peroxidase 4 （GPX4）/ solute carrier family 7 member 11 （SLC7A11） pathway
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2026;32(5):101-113
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveThe paper aims to investigate the action mechanism by which the Xiaozheng Zhitong paste （XZP） relieves bone cancer pain （BCP）. MethodsA model of mice with BCP was established by using Lewis tumor cells. The therapeutic effects of XZP， the ferroptosis inhibitor Ferrostatin-1 （Fer-1）， and the nuclear factor erythroid 2-related factor 2 （Nrf2） inhibitor Brusatol （Bru） on BCP were examined. Mice were randomly divided into the Sham operation group， BCP group， BCP+XZP-L group， BCP+XZP-H group， BCP+Fer-1 group， and BCP+XZP-H+Bru group， with six mice in each group. Pain behavior tests were conducted on the mice to assess pain levels. Colorimetric assays were employed to measure ferroptosis-related factors in serum and spinal cord tissue including Fe， malondialdehyde （MDA）， reactive oxygen species （ROS）， and superoxide dismutase （SOD）. Immunofluorescence staining was used to assess ROS production in spinal cord tissue. Transmission electron microscopy was used to observe the ultrastructure of mitochondria in lumbar spinal cord tissue. Quantitative real-time polymerase chain reaction （Real-time PCR） was employed to detect mRNA expression of Nrf2， heme oxygenase-1 （HO-1）， glutathione peroxidase 4 （GPX4）， and solute carrier family 7 member 11 （SLC7A11） in spinal cord neuron tissue. The protein expression of Nrf2， HO-1， GPX4， and SLC7A11 in spinal cord neurons was measured by Western blot. ResultsCompared with the Sham group， mice in the BCP group exhibited significantly reduced limb usage scores， mechanical foot withdrawal thresholds， and thermal foot withdrawal thresholds （P<0.01）. Serum and lumbar spinal cord tissue levels of Fe， MDA， and reactive oxygen species （ROS） were significantly elevated （P<0.05）， while superoxide dismutase （SOD） levels were significantly decreased （P<0.05）. Lumbar spinal cord mitochondrial structural damage was observed， and mRNA and protein expression of Nrf2， HO-1， GPX4， and SLC7A11 were significantly downregulated （P<0.01）. Compared with the BCP group， both low- and high-dose XZP groups improved the aforementioned pain behavioral indicators （P<0.05，P<0.01）， reduced ferroptosis-related biomarkers including Fe， MDA， and ROS levels （P<0.05）， increased SOD levels （P<0.05，P<0.01）， alleviated mitochondrial damage， and upregulated Nrf2， HO-1， GPX4， SLC7A11 mRNA and protein expression （P<0.05，P<0.01）. The high-dose XZP group exhibited comparable efficacy to Fer-1 in alleviating pain and inhibiting ferroptosis. Following Bru administration， XZP's effects on pain behavioral indicators， regulation of ferroptosis-related markers， mitochondrial structural protection， and activation of the Nrf2/HO-1/GPX4/SLC7A11 pathway were significantly reversed （P<0.05，P<0.01）. ConclusionExternal application of XZP alleviates pain symptoms in BCP mice by activating the Nrf2/HO-1/GPX4/SLC7A11 pathway， thereby inhibiting ferroptosis and neuronal damage in spinal cord neurons.
