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Journal of Chinese Integrative Medicine ›› 2007, Vol. 5 ›› Issue (1): 39-44.doi: 10.3736/jcim20070108

• Original Experimental Research • Previous Articles     Next Articles

A rabbit model of cervical spondylosis established by stimulation of wind, cold and dampness

Yong-jun Wang, Qi Shi, Quan Zhou, Chen-guang Li, Jiang-ming Cheng, Mei Liu, Yang Yang, Chong-jian Zhou   

  1. Institute of Spine, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China
  • Online:2007-01-31 Published:2007-01-21

Objective: To make an animal model of cervical spondylosis (arthralgia syndrome type) with stimulation of wind, cold, and dampness. 

Methods: Twenty-four 8 months old male New Zealand white rabbits were randomly allocated into four groups: normal control group, light stimulation group, moderate stimulation group and severe stimulation group. The wind speed was 10.8-13.8 m/s, the temperature was (5±0.5) ℃, and the humidity was 100%. The rabbits of light, moderate, and severe stimulation groups were kept in the above-mentioned environments for 4 hours everyday, and for a total of 32, 64, and 128 hours, respectively. The intervertebral discs were stained with HE method, and observed with a light microscope. Prostaglandin E2 (PGE2), 6-ketone-prostaglandin F (6-K-PGF) and thromboxane B2 (TXB2) contents were measured by ELISA. Fas and Bcl-2 expressions were examined by immunohistochemical avidin-biotin peroxidose complex technique. Interleukin-1β (IL-1β), tumor necrosis factor α (TNF-α), and transforming growth factor β (TGF-β) mRNA expressions were examined by reverse transcription-polymerase chain reaction.

Results: The nucleus pulposus of rabbits in the light and moderate stimulation groups shrunken, and in the severe stimulation group, the anulus fibrosus loosed or ruptured, and the cartilage end-plate became proliferated. Compared with rabbits in the normal control group, the PGE2 content rose in the light stimulation group, the contents of PGE2, 6-K-PGF, and TXB2 increased, the expressions of IL-1β and TNF-α mRNAs and Fas were up-regulated, and the expressions of TGF-β mRNA and Bcl-2 were down-regulated in the moderate and severe stimulation groups. The expression of Fas was up-regulated mostly and Bcl-2 was down-regulated mostly in the severe group. 

Conclusion: Moderate and severe stimulations of wind, cold and dampness can lead to degeneration of cervical intervertebral discs of rabbits. The model corresponds to the theory of traditional Chinese medicine about arthralgia syndrome caused by wind, cold and dampness.

Key words: cervical vertebrae, wind-cold-dampness, arthralgia syndrome, rabbit

CLC Number: 

  • R681.5

Figure 1

SHH-250GS improved artificial climate mode maker boxStimulus intensity: wind force, 6 degree; temperature, (5±0.5) ℃; humidity, 100%. Rabbits of light, moderate, and severe stimulation groups were stimulated for 4 hours everyday, and for a total of 32, 64, and 128 hours, respectively."

Figure 2

Morphological observation of intervertebral discs (HE staining, ×10)A: Normal control group; B: Light stimulation group; C: Moderate stimulation group; D: Severe stimulation group. The disc structure of the normal control group was fine, in which annular fibrosus and nucleus pulposus were fine arranged. The disc nucleus pulposus of both light and moderate stimulation groups was shrunk, and the annular fibrosus was loosened. The disc annular fibrosus of the severe stimulation group was loosen and broken, and the cartilage end-plate was thickened."

Table 1

Contents of PGE2, 6-K-PGF1α and TXB2 in intervertebral discs ($\bar{x}±S$ ,pg/mg)"

Group n PGE2 6-K-PGF TXB2
Normal control 6 61.36±6.34 746.23±223.13 216.45±68.11
Light stimulation 6 396.71±24.26** 978.19±235.46 429.82±131.26
Moderate stimulation 6 550.56±83.77** 1 084.06±182.23 619.23±205.62
Severe stimulation 6 438.72±66.06** 1 250.44±281.21* 955.80±203.35**

Table 2

OD value ratios of IL-1β, TNF-α and TGF-β mRNAs in intervertebral discs ($\bar{x}±S$)"

Group n IL-1β mRNA TNF-α mRNA TGF-β mRNA
Normal control 6 0.852 3±0.026 4 0.890 3±0.010 1 1.152 9±0.044 2
Light stimulation 6 0.915 3±0.006 1** 0.962 3±0.012 8** 1.054 9±0.015 8**
Moderate stimulation 6 0.924 5±0.002 3** 0.981 0±0.071 4** 0.931 9±0.014 0**△△
Severe stimulation 6 0.959 6±0.005 7**△△▲ 1.010 2±0.010 7** 0.862 5±0.035 2**△△

Figure 3

Expression of Fas in four groups (Immunohistochemical staining, ×200)A: Normal control group; B: Light stimulation group; C: Moderate stimulation group; D: Severe stimulation group."

Figure 4

Expression of Bcl-2 in four groups (Immunohistochemical staining, ×200)A: Normal control group; B: Light stimulation group; C: Moderate stimulation group; D: Severe stimulation group."

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