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Journal of Chinese Integrative Medicine ›› 2008, Vol. 6 ›› Issue (10): 1034-1039.doi: 10.3736/jcim20081009

• Original Experimental Research • Previous Articles     Next Articles

Establishment of a rat model of cervical syndrome with kidney deficiency

Jian-chun Jiang1, Chen-guang Li1, Qian-qian Liang1, Qin Bian1, Quan Zhou1, Xue-jun Cui2, Min Huang1, Qing-gao Liu1,Sheng Lu1, Chong-jian Zhou1, Qi Shi1, Yong-jun Wang1()   

  1. 1. Institute of Spine, 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:2008-05-30 Online:2008-10-20 Published:2008-10-15

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Objective:

To establish a rat model of cervical syndrome (CS) with kidney deficiency.


Methods: 

A group of 30 three-month-old female Sprague-Dawley rats were randomly divided into normal control group, CS group and CS with kidney deficiency group (combined group), with 10 rats in each group. Rats in the normal control group received no treatment, rats in the CS group underwent only 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 as kidney deficiency model. Serum and cervical intervertebral discs were collected. Kidney deficiency was determined by observing the morphologic changes of uterus and appendages, detecting the weight of uterus and appendages and the content of serum estradiol (E2). The degeneration of intervertebral discs was determined by detecting the histopathology, the expressions of type Ⅱcollagen and type Ⅹcollagen proteins, and the expressions of aggrecan-1 (Agc1), type Ⅱ procollagen gene (Col2a1), matrix metalloproteinase-13 (MMP-13) and tissue inhibitor of metalloproteinases-1 (TIMP-1) mRNAs.


Results:

Compared with those in the normal control group and CS group, the rats in the combined group were noted with the uterus atrophied, the caliber of oviduct thinned, the weight of uterus and appendages diminished obviously, the content of serum E2 decreased, cervical intervertebral disc degenerated more seriously, type Ⅱ collagen protein expression decreased, type Ⅹ collagen protein expression increased, Agc1 and Col2a1 mRNA expressions in intervertebral disc decreased, and the MMP-13 mRNA expression increased.


Conclusion:

The rat model of CS with kidney deficiency is established. Kidney deficiency can aggravate cervical intervertebral disc degeneration.

Key words: kidney deficiency, cervical syndrome, combining disease and syndrome, disease model, animal, rats

CLC Number: 

  • R681.531

Table 1

The primer sequences of Agc1、Col2a1、MMP-13 and TIMP-1"

Target gene Primer sequence Fragment size (bp)
β-actin Forward: 5'-GGA GAT TAC TGC CCT GGC TCC TA-3' 150
Reserve: 5'-GAC TCA TCG TAC TCC TGC TTG CTG-3'
Agc1 Forward: 5'-TCC GCT GGT CTG ATG GAC AC-3' 101
Reserve: 5'-AGG ACC AAC TTT GCC TTG AGG AC-3'
Col2a1 Forward: 5'-TCC TAA GGG TGC CAA TGG TGA-3' 112
Reserve: 5'-AAT GTC AAC AAT GGG AAG GCG T-3'
MMP-13 Forward: 5'-CCC TGG AGC CCT GAT GTT T-3' 142
Reserve: 5'-CTC TGG TGT TTT GGG GTG CT-3'
TIMP-1 Forward: 5'-ACA GGT TTC CGG TTC GCC TAC-3' 134
Reserve: 5'-CTG CAG GCA GTG ATG TGC AA-3'

Table 2

Weight of uterus and appendages in three groups (x±s, g)"

Group n Weight of uterus and appendages
Normal control 8 0.94±0.19
Cervical syndrome 8 0.98±0.17
Combined 8 0.18±0.02**△△

Table 3

Content of serum E2 in three groups (x±s, μg/L)"

Group n E2
Normal control 8 3.15±0.68
Cervical syndrome 8 2.69±0.32
Combined 8 2.46±0.40*

Figure 1

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

Table 4

Miyamoto scores in three groups (x±s)"

Group n Miyamoto score
Normal control 8 1.38±0.52
Cervical syndrome 8 2.25±0.46*
Combined 8 4.00±0.53**△△

Figure 2

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

Table 5

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

Group n Agc1/β-actin Col2a1/β-actin MMP-13/β-actin TIMP-1/β-actin
Normal control 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** 0.32±0.18* 2.72±0.48**△△ 0.55±0.15
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