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Journal of Traditional Chinese Medicine >

2023 , Vol. 43 >Issue 5: 991 - 1000

DOI: https://doi.org/10.19852/j.cnki.jtcm.2023.05.005

Effect of Tuina along “bladder meridian” alleviating intervertebral disc degeneration by regulating the transforming growth factor-β1/Smad signaling pathway in a rabbit model

  • Chengguo SU ,
  • Xiaoyan ZHAO ,
  • Jiangnan YE ,
  • Xin ZHANG ,
  • Yuqing JIANG ,
  • Junjie GUO ,
  • Xiyuan ZHANG ,
  • Wenchuan QI ,
  • Jun ZHU
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  • 1 Acupuncture and Tuina School, Chengdu University of Traditional Chinese Medicine, Chengdu 610075, China
    2 Clinical Medical School, Affiliated Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu 610075, China

Received date: 2022-03-23

  Accepted date: 2022-07-15

  Online published: 2023-08-28

Supported by

National Natural Science Foundation of China: Based on TGF-β1/Smads Signaling Pathway to Study the Effect Mechanism of Tuina along the Bladder Meridian on Intervertebral Disc Degeneration(82004497);China Postdoctoral Science Foundation: Based on TGF-β1/RhoA/JNK Signaling Pathway to Study the Effect Mechanism of Tuina along the Bladder Meridian on Intervertebral Disc Degeneration(2021M693788)

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Abstract

OBJECTIVE: The aim of this study was to investigate the protective effects of Tuina (a traditional Chinese massage therapy) on intervertebral disc (IVD) degeneration and the regulatory mechanisms of the transforming growth factor-β1 (TGF-β1)/small mothers against decapentaplegic (Smad) signaling pathway.
METHODS: Thirty New Zealand white rabbits were randomized into five groups: the control group, model group, model + Tuina group (Tuina group), model + TGF-β1 group (TGF-β1 group), and model + TGF-β1 inhibitor SB431542 group (SB431542 group). The model was established by posterolateral annulus fibrosus puncturing (AFP). Recombinant TGF-β1 and inhibitor SB431542 was injected into the TGF-β1 group and SB431542 group with a microsyringe, respectively. The rabbits in the Tuina group received Tuina treatment along the bladder meridian for 4 weeks. Magnetic resonance imaging (MRI) was performed on rabbits before AFP and after 4 weeks of intervention. Lumbar IVDs (L2-L3 to L4-L5) were harvested after intervention. Histopathological changes in the IVDs were measured by hematoxylin and eosin (HE) staining. Type I collagen was analyzed by immunohistochemistry detection. The expression level of matrix metalloproteinase-3 (MMP3) was determined by enzyme-linked immunosorbent assay. Cell apoptosis was evaluated by terminal deoxynucleotidyl transferase-mediated nick end labeling and Western blotting. Real-time polymerase chain reaction and Western blotting were used to analyze the expression of TGF-β1 and Smad2/3/4 and a disintegrin and metalloproteinase with thrombospondin motifs 5.
RESULTS: Posterolateral AFP induced IVD degeneration in rabbits with histopathological damage and noticeable changes in MRI images. Tuina alleviated histo-pathological changes and reversed the expression of extracellular matrix degeneration-related molecules and apoptosis-related proteins. Furthermore, AFP induced the activation of TGF-β1 and Smad2/3/4, whereas Tuina therapy markedly reduced the protein expression of Smad2/3 and the gene expression of TGF-β1 and Smad2/3/4. Additionally, the TGF-β1/Smad signaling pathway was activated in the TGF-β1 group, while the TGF-β1/Smad signaling pathway was inhibited in the SB431542 group.
CONCLUSION: Posterolateral AFP induced disc degeneration as determined by MRI assessment and histological analysis. Tuina alleviated disc degeneration, possibly by inhibiting the fibrotic response mediated by the TGF-β1/Smad pathway, thus alleviating extracellular matrix degeneration and reducing cell apoptosis.

Key words： intervertebral disc degeneration; transforming growth factor beta1; Smad proteins; Tuina; apoptosis; extracellular matrix degradation

Cite this article

Chengguo SU , Xiaoyan ZHAO , Jiangnan YE , Xin ZHANG , Yuqing JIANG , Junjie GUO , Xiyuan ZHANG , Wenchuan QI , Jun ZHU . Effect of Tuina along “bladder meridian” alleviating intervertebral disc degeneration by regulating the transforming growth factor-β1/Smad signaling pathway in a rabbit model[J]. Journal of Traditional Chinese Medicine, 2023 , 43(5) : 991 -1000 . DOI: 10.19852/j.cnki.jtcm.2023.05.005

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