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Journal of Chinese Integrative Medicine ›› 2006, Vol. 4 ›› Issue (6): 615-619.doi: 10.3736/jcim20060615

• Original Experimental Research • Previous Articles     Next Articles

Study of Yiqi Huayu Bushen Recipe and its decomposed formulas in regulating gene expressions in degenerated cervical intervertebral discs of rats

Yong-jun Wang,Qi Shi,Chen-guang Li,Quan Zhou,Zhi-jun Hu,Mei Liu,Chong-jian Zhou   

  1. Institute of Spine, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China
  • Online:2006-11-30 Published:2006-11-15

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Objective

To investigate the gene expression changes in the degenerated cervical intervertebral discs of rats, and to study the function of Yiqi Huayu Bushen Recipe, a compound Chinese herbal medicine, and its discomposed formulas in regulating gene expressions in the degenerated cervical intervertebral discs.

Methods

The rat model with degenerated cervical intervertebral discs caused by imbalance between the dynamic and static forces was established. The mRNA was extracted from the cervical intervertebral discs of rats in the normal control and experiment groups, and the cDNA probes were obtained by inverse transcript. The cDNA probes were hybridized with the gene chips. The gene expression pattern was gained with a laser scanner. Image analysis, standardized ratio value and cluster analysis were used to investigate the differential of gene expressions between the control and experiment groups.

Results

Cluster analysis showed that the gene chips of No.1 (Yiqi Huayu group), No.2 (Yiqi Bushen group) and No.3 (Huayu Bushen group) were in one class, while the gene chips of No.4 (untreated group) and No.5 (Yiqi Huayu Bushen group) were in the other. The gene expression of No.4 was different from the others mostly, and the gene expressions of No.2 and No.3 were similar. There were 96 genes expressed differently in three cases and among them 77 genes were already known and the expression of 48 genes were up-regulated (ratio>1.0), and 29 down-regulated (ratio＜0.5). There were 25 genes expressed differently between the untreated group and the herb-treated groups. 

Conclusions

The gene expressions of the degenerated rat intervertebral discs are changed. Yiqi Huayu Bushen Recipe and its discomposed formulas have the effect of regulating the expressions of related genes, such as PI3K, PTK, ERK3, and PH1B1.

Key words: Cervical vertebrae, Gene chip, Cluster analysis, Signal transduction, Rats

CLC Number: 

  • R289.5

Table 1

Numbering and grouping of gene chips"

Gene Chip NumberSample GroupFluorescent marker
1Untreated groupcy3（control group）
Yiqi Huayu Recipe-treated groupcy5（experiment group）
2Untreated groupcy3（control group）
Yiqi Bushen Recipe-treated groupcy5（experiment group）
3Untreated groupcy3（control group）
Huayu Bushen Recipe-treated groupcy5（experiment group）
4Untreated groupcy3（control group）
Sham-operated groupcy5（experiment group）
5Untreated groupcy3（control group）
Yiqi Huayu Bushen Recipe-treated groupcy5（experimental group）

Figure 1

Folded figure of double fluorescent labeling of No.4 gene chipThe green spots represent the cy3 signal is stronger (down regulated), the red spots represent the cy5 signal is stronger (up regulated), and the yellow spots represent the two signals of cy3 and cy5 were in similar intensity."

Figure 2

Scattered plots of cross-fertilization signals of No.4 gene chipThe X and Y axes represent the fluorescence intensities of cy3 and cy5 signals separately. Every spot responds to the cross-fertilization signal of one gene. The red spots represent ratios between 0.5 and 1.0, which mean no difference in expression; the yellow spots represent ratio>1.0 or ratio＜0.5, which mean difference in expression."

Table 2

Classification of gene expressions by cluster analysis and the gene ID"

Different typeGene ID
Expression in Chinese medicine treated group up-regulated, and expression in untreated group no differencen0128c11, n0085e08, n0091g09, n0255a01, n0233h11, n0003e11, n0290c09
Expression in Chinese medicine treated group up-regulated, and expression in untreated group down regulatedn0011b03, n0096h01, n0021e01, n0018h09, n0007c08, n0158a09, n0135e05, n0095a08, n0320e05
Expression in Chinese medicine treated group down-regulated, and expression in untreated group no differencen0289e07, n0301e10, n0243g02, n0056h11, n0033f05, n0256g11, n0126d12,
Expression in Chinese medicine treated group down-regulated, and expression in untreated group up-regulatedn0037a10, n0022b02

Figure 2

Treeveiw of cluster analysis"

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