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[Altered expression of endogenous transforming growth factor β1 and early calcification related genes in rat endplate].
Zhonghua Yi Xue Za Zhi 2011; 91(31):2181-5ZY

Abstract

OBJECTIVE

To explore the relationship between endogenous transforming growth factor (TGF)-β1 and calcification-related genes through an in vitro degeneration model by propagating rat endplate chondrocytes during a natural degeneration process.

METHODS

Endplate chondrocytes were extracted from rat lumbar vertebrae, isolated by enzyme digestion and P2 and P4 generations selected for a 6-day in vitro culture. The specimens were photographed microscopically to observe the cellular differences by alizarin red staining. Type II collagen marker gene, transcription factor SOX-9 gene and metabolism-related genes proteoglycan. matrix metalloproteinase (MMP)-13, a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS)-4 and ADAMTS-5 were detected by RT-PCR to verify the degeneration model. Based on this model, the changes of growth factor TGF-β1 and calcification-related genes ankyrin (ANK), ectonucleotide pyrophosphatase (ENPP), tissue-nonspecific alkaline phosphatase (TNAP) were continuously tested.

RESULTS

Compared with P2 cells, P4 cells tended to assume a spindle-shaped morphology. And alizarin red staining showed no change between them. The level of transcription factor SOX-9 of P4 cells (P4/P2 = 0.0690, P = 0.0489) was significantly lower than that of P2 cells. Type II collagen (P4/P2 = 0.0535, P = 0.009) and proteoglycan (P4/P2 = 0.2672, P = 0.0343) were also significantly lower than those of P2 cells. No significant changes were observed in other metabolism-related genes. TGF-β1 (P4/P2 = 0.5934, P = 0.0482) was significantly lower. The expressions of TNAP (P4/P2 = 0.0385, P = 0.0139) and ANK (P4/P2 = 0.2121, P = 0.0009) were significantly lower. But ENPP showed no significant change.

CONCLUSION

P4 endplate chondrocytes undergo natural degeneration in vitro with the rising passage number. Type II collagen, SOX-9 and proteoglycan are significantly reduced. Endogenous TGF-β1 gene and calcification-related genes are down-regulated. The decrease of ANK gene may be caused by the down-regulation of endogenous TGF-β1. Modulating the expression of endogenous TGF-β1 gene in endplate chondrocytes may become a new therapeutic approach for the degeneration of intervertebral disc.

Authors+Show Affiliations

Department of Orthopedic Surgery, Yijishan Hospital, Wannan Medical College, Wuhu 241001, China. xuhg@medmail.com.cnNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

English Abstract
Journal Article
Research Support, Non-U.S. Gov't

Language

chi

PubMed ID

22094034

Citation

Xu, Hong-Guang, et al. "[Altered Expression of Endogenous Transforming Growth Factor Β1 and Early Calcification Related Genes in Rat Endplate]." Zhonghua Yi Xue Za Zhi, vol. 91, no. 31, 2011, pp. 2181-5.
Xu HG, Zhang XL, Zhang XH, et al. [Altered expression of endogenous transforming growth factor β1 and early calcification related genes in rat endplate]. Zhonghua Yi Xue Za Zhi. 2011;91(31):2181-5.
Xu, H. G., Zhang, X. L., Zhang, X. H., Wang, H., Jia, R. P., Tong, W. X., & Tong, H. J. (2011). [Altered expression of endogenous transforming growth factor β1 and early calcification related genes in rat endplate]. Zhonghua Yi Xue Za Zhi, 91(31), pp. 2181-5.
Xu HG, et al. [Altered Expression of Endogenous Transforming Growth Factor Β1 and Early Calcification Related Genes in Rat Endplate]. Zhonghua Yi Xue Za Zhi. 2011 Aug 23;91(31):2181-5. PubMed PMID: 22094034.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - [Altered expression of endogenous transforming growth factor β1 and early calcification related genes in rat endplate]. AU - Xu,Hong-Guang, AU - Zhang,Xiao-Ling, AU - Zhang,Xiao-Hai, AU - Wang,Hong, AU - Jia,Rui-Ping, AU - Tong,Wen-Xue, AU - Tong,Hai-Jun, PY - 2011/11/19/entrez PY - 2011/11/19/pubmed PY - 2012/4/24/medline SP - 2181 EP - 5 JF - Zhonghua yi xue za zhi JO - Zhonghua Yi Xue Za Zhi VL - 91 IS - 31 N2 - OBJECTIVE: To explore the relationship between endogenous transforming growth factor (TGF)-β1 and calcification-related genes through an in vitro degeneration model by propagating rat endplate chondrocytes during a natural degeneration process. METHODS: Endplate chondrocytes were extracted from rat lumbar vertebrae, isolated by enzyme digestion and P2 and P4 generations selected for a 6-day in vitro culture. The specimens were photographed microscopically to observe the cellular differences by alizarin red staining. Type II collagen marker gene, transcription factor SOX-9 gene and metabolism-related genes proteoglycan. matrix metalloproteinase (MMP)-13, a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS)-4 and ADAMTS-5 were detected by RT-PCR to verify the degeneration model. Based on this model, the changes of growth factor TGF-β1 and calcification-related genes ankyrin (ANK), ectonucleotide pyrophosphatase (ENPP), tissue-nonspecific alkaline phosphatase (TNAP) were continuously tested. RESULTS: Compared with P2 cells, P4 cells tended to assume a spindle-shaped morphology. And alizarin red staining showed no change between them. The level of transcription factor SOX-9 of P4 cells (P4/P2 = 0.0690, P = 0.0489) was significantly lower than that of P2 cells. Type II collagen (P4/P2 = 0.0535, P = 0.009) and proteoglycan (P4/P2 = 0.2672, P = 0.0343) were also significantly lower than those of P2 cells. No significant changes were observed in other metabolism-related genes. TGF-β1 (P4/P2 = 0.5934, P = 0.0482) was significantly lower. The expressions of TNAP (P4/P2 = 0.0385, P = 0.0139) and ANK (P4/P2 = 0.2121, P = 0.0009) were significantly lower. But ENPP showed no significant change. CONCLUSION: P4 endplate chondrocytes undergo natural degeneration in vitro with the rising passage number. Type II collagen, SOX-9 and proteoglycan are significantly reduced. Endogenous TGF-β1 gene and calcification-related genes are down-regulated. The decrease of ANK gene may be caused by the down-regulation of endogenous TGF-β1. Modulating the expression of endogenous TGF-β1 gene in endplate chondrocytes may become a new therapeutic approach for the degeneration of intervertebral disc. SN - 0376-2491 UR - https://www.unboundmedicine.com/medline/citation/22094034/[Altered_expression_of_endogenous_transforming_growth_factor_β1_and_early_calcification_related_genes_in_rat_endplate]_ L2 - http://journal.yiigle.com/LinkIn.do?linkin_type=pubmed&issn=0376-2491&year=2011&vol=91&issue=31&fpage=2181 DB - PRIME DP - Unbound Medicine ER -