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Journal of Chinese Integrative Medicine ›› 2008, Vol. 6 ›› Issue (11): 1152-1158.doi: 10.3736/jcim20081109

• Original Experimental Research • Previous Articles     Next Articles

Establishment of a rat model of cervical syndrome with qi deficiency, blood stasis and kidney deficiency

Yong-jun Wang1(), Qi Shi1, Jian-chun Jiang1, Qian-qian Liang1, Qin Bian1, Chen-guang Li1, Quan Zhou1, Xue-jun Cui1, Min Huang2, Qing-gao Liu1, Sheng Lu1, Chong-jian Zhou1   

  1. 1. Institute of Spine Disease, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China
    2. Health Service Center of the Bund Community, Huangpu District, Shanghai 200001, China
  • Received:2008-05-27 Online:2008-11-20 Published:2008-11-15

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Objective: To establish a rat model of cervical syndrome with qi deficiency, blood stasis and kidney deficiency.
Methods: A total of 30 three-month-old female Sprague-Dawley rats were randomly divided into normal group, cervical syndrome group and cervical syndrome with qi deficiency, blood stasis and kidney deficiency group (combined group), with 10 rats in each group. Rats in the normal group received no treatment, rats in cervical syndrome group underwent resection of cervical muscles and ligaments as unbalanced dynamic and static animal model, and rats in combined group underwent resection of both cervical muscles and ovaries, swimming and irregular diet, and injection of adrenal cortex hormone and adrenaline two and a half months after resection as combined model. The qi deficiency, blood stasis and kidney deficiency were determined by observing behaviors and physical signs of the rats, detecting the contents of plasma cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP), the hemorrheology, the expression of alpha-granular membrane protein (CD62p) and the serum estradiol (E2) content. The aggrecan-1, type Ⅱ procollagen gene (Col2a1), matrix metalloproteinases-13 (MMP-13) and tissue inhibitor of metalloproteinases-1 (TIMP-1) mRNAs in cervical intervertebral discs were detected by histopathology, immunohistochemistry and real-time polymerase chain reaction. The cataplasia of the intervertebral discs was determined by detecting the histopathology, the expressions of type Ⅱ collagen and type Ⅹ collegen proteins, and the expressions of aggrecan-1 (Agc1), type Ⅱ procollagen gene (Col2a1), matrix metalloproteinase-13 (MMP-13) and tissue inhibitor of metalloproteinase-1 (TIMP-1) mRNAs.
Results: Compared with those in the normal group and cervical syndrome group, the rats in the combined group were noted with obvious signs of deficiency of vital energy, such as depression, tiredness, ptosis, obvious weight loss and blue tail. And the ratio of cAMP/cGMP was decreased; the reducing viscosity was significantly up-regulated; the expression of CD62p was increased; the content of serum E2 was decreased; the intervertebral disc structure was destructed; the cervical intervertebral disc was more seriously deteriorated. There exhibited a decrease in type Ⅱ collagen protein expression, an increase in type Ⅹ collagen protein expression, as well as decreases of Agc1, Col2a1 and TIMP-1 mRNA expressions in intervertebral disc, and the expression of MMP-13 mRNA was noted an increase.
Conclusion: The rat model of cervical syndrome with qi-deficiency, blood stasis and kidney deficiency is established. Qi deficiency, blood stasis and kidney deficiency can aggravate cervical intervertebral disc degeneration.

Key words: qi deficiency, blood stasis, kidney deficiency, cervical spondylosis, combining disease and syndrome, disease model, animals, rats

CLC Number: 

  • R681.531

Table 1

Observation of behaviors and physical signs in the combined group when making qi deficiency model"

Symptom Combined group
3rd day 6th day 9th day 13th day 15th day
Spirit + ++ +++ +++ +++
Skin and pelage + + ++ ++ +++
Nose and tail - + ++ ++ ++
Stool - + + + +
Shrinking shoulders and extrados none none have have have

Table 2

Weight changes in the combined group when making qi deficiency model(x±s, g)"

Group n Body weight
Before making model 3rd day 6th day 9th day 13th day 15th day
Normal 10 328.9±28.8 340.0±16.0 343.0±18.3 347.5±17.2 349.3±19.1 353.7±19.6
Cervical syndrome 10 325.5±21.5 341.3±16.9 342.2±18.9 350.0±20.5 351.9±26.3 357.0±23.1
Combined 8 431.1±31.2**△△ 421.7±55.1**△△ 389.0±46.4**△△ 387.5±58.1 387.9±54.8 358.9±52.8

Table 3

Contents of cAMP, cGMP and ratio of cAMP/cGMP in three groups (x±s)"

Group n cAMP (nmol/L) cGMP (nmol/L) cAMP/cGMP
Normal 8 129.35±60.52 17.87±9.51 8.10±3.27
Cervical syndrome 8 217.51±117.37 23.36±19.91 11.97±4.56
Combined 8 165.06±56.56 44.28±32.88 4.77±1.77△△

Table 4

Changes of hemorrheological parameters in three groups (x±s, mPa·s)"

Group n Whole blood low-shear viscosity Plasma viscosity Reducing blood viscosity Aggregation index
Normal 8 13.22±1.38 1.09±0.02 25.68±2.22 2.30±0.15
Cervical syndrome 8 12.70±1.06 1.12±0.08 24.99±1.72 2.30±0.12
Combined 8 13.85±1.24 1.37±0.38 27.81±2.13*△△ 2.58±0.15

Table 5

Expression of CD62p and content of serum E2 in three groups (x±s)"

Group n CD62p (%) E2 (μg/L)
Normal 8 2.89±0.90 3.15±0.68
Cervical syndrome 8 8.16±1.85 2.69±0.32
Combined 8 51.80±11.50**△△ 1.12±0.83**△△

Figure 1

HE staining of cervical intervertebral disc (Light microscopy, ×40) A: Normal group; B: Cervical syndrome group; C: Combined group"

Figure 2

Immunohistochemical staining of type Ⅱ collagen in annular fibrosus of cervical intervertebral disc (Light microscopy, ×200) A: Normal group; B: Cervical syndrome group; C: Combined group."

Figure 3

Immunohistochemical staining of type Ⅹ collagen of cervical intervertebral disc (Light microscopy, ×200) A: Normal group; B: Cervical syndrome group; C: Combined group."

Table 6

Expressions of Agc1, Col2a1, MMP-13 and TIMP-1 mRNAs of cervical intervertebral disc in three groups（x±s）"

Group n Agc1/β-actin Col2a1/β-actin MMP-13/β-actin TIMP-1/β-actin
Normal 6 1.00±0.31 1.00±0.34 1.00±0.25 1.00±0.43
Cervical syndrome 6 0.06±0.04** 0.58±0.01 1.69±0.30* 0.41±0.08
Combined 6 0.02±0.01** 0.30±0.20* 2.25±0.21**△ 0.76±0.24
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