Leg amputation accelerates senescence of rat lumbar intervertebral discs

Qiu Juan Xing, Qian Qian Liang, Qin Bian, Dao Fang Ding, Xue Jun Cui, Qi Shi, Yong Jun Wang

Research output: Contribution to journalArticle

Abstract

Study Design: Several senescence biomarkers were observed to investigate cell senescence in degenerative intervertebral lumbar discs of foreleg-amputated rats. Objective: To determine if cell senescence is accelerated in degenerative intervertebral lumbar disc cells in an upright-rat model. Summary Of Background Data: Cellular senescence was accelerated in human and sand rat degenerative intervertebral disc (IVD) cells. Repeated use of upright posture by rats contributed to degenerative disc changes. No convincing evidence of cell senescence was observed in the lumbar disc of the foreleg amputated rat. Methods: The forelimbs of 20 rats were amputated at 1 month of age such that they maintained an upright stance; rats were housed in custom-made cages. Nonamputated rats, also 1 month of age, were kept in regular cages and served as a control group. The lumbar IVDs were harvested from rats in 2 groups, at 5 or 9 months following amputation. Senescence-associated-β-galactosidase- positive staining was used to detect cell senescence. p16INK4a and p27KIP were assessed by immunohistochemistry analysis. Total RNA isolated from these samples was used to measure the gene expression of p16INK4a, RB, cyclin D1, CDK4, PTEN, p27KIP, p19ARF, p21, TERT, and RAGE by real-time polymerase chain reaction assay. Results: The highest levels of SA-β-GAL activity were detected in 9-month amputated rats. Quantitative immunohistochemical analysis showed that there were highest rates of p16INK4a and p27KIP protein expression in the cartilage endplate and anulus fibrosus of 9-month amputated rats. The mRNA levels of p16INK4a, RB, PTEN, p27KIP, p19ARF, and RAGE were upregulated. The increased cyclin D1 mRNA level was statistically significant only at the ninth month following amputation; CDK4 and TERT mRNA levels were downregulated to a similar extent at both points compared with nonamputated controls. mRNA expression of p21 was significantly downregulated. Conclusion: Accelerated cell senescence was associated with forelimb amputation that causes abnormal loading in rat lumbar IVDs.

Original languageEnglish (US)
JournalSpine
Volume35
Issue number23
DOIs
StatePublished - Nov 1 2010
Externally publishedYes

Fingerprint

Intervertebral Disc
Amputation
Leg
Cell Aging
Intervertebral Disc Degeneration
Messenger RNA
Forelimb
Cyclin D1
Down-Regulation
Galactosidases
Cyclin-Dependent Kinase Inhibitor p16
Gerbillinae
Posture
Cartilage
Real-Time Polymerase Chain Reaction
Biomarkers
Immunohistochemistry
RNA
Staining and Labeling
Gene Expression

Keywords

  • intervertebral disc
  • lumbar
  • senescence

ASJC Scopus subject areas

  • Orthopedics and Sports Medicine
  • Clinical Neurology

Cite this

Xing, Q. J., Liang, Q. Q., Bian, Q., Ding, D. F., Cui, X. J., Shi, Q., & Wang, Y. J. (2010). Leg amputation accelerates senescence of rat lumbar intervertebral discs. Spine, 35(23). https://doi.org/10.1097/BRS.0b013e3181e7d087

Leg amputation accelerates senescence of rat lumbar intervertebral discs. / Xing, Qiu Juan; Liang, Qian Qian; Bian, Qin; Ding, Dao Fang; Cui, Xue Jun; Shi, Qi; Wang, Yong Jun.

In: Spine, Vol. 35, No. 23, 01.11.2010.

Research output: Contribution to journalArticle

Xing, QJ, Liang, QQ, Bian, Q, Ding, DF, Cui, XJ, Shi, Q & Wang, YJ 2010, 'Leg amputation accelerates senescence of rat lumbar intervertebral discs', Spine, vol. 35, no. 23. https://doi.org/10.1097/BRS.0b013e3181e7d087
Xing, Qiu Juan ; Liang, Qian Qian ; Bian, Qin ; Ding, Dao Fang ; Cui, Xue Jun ; Shi, Qi ; Wang, Yong Jun. / Leg amputation accelerates senescence of rat lumbar intervertebral discs. In: Spine. 2010 ; Vol. 35, No. 23.
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AU - Liang, Qian Qian

AU - Bian, Qin

AU - Ding, Dao Fang

AU - Cui, Xue Jun

AU - Shi, Qi

AU - Wang, Yong Jun

PY - 2010/11/1

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N2 - Study Design: Several senescence biomarkers were observed to investigate cell senescence in degenerative intervertebral lumbar discs of foreleg-amputated rats. Objective: To determine if cell senescence is accelerated in degenerative intervertebral lumbar disc cells in an upright-rat model. Summary Of Background Data: Cellular senescence was accelerated in human and sand rat degenerative intervertebral disc (IVD) cells. Repeated use of upright posture by rats contributed to degenerative disc changes. No convincing evidence of cell senescence was observed in the lumbar disc of the foreleg amputated rat. Methods: The forelimbs of 20 rats were amputated at 1 month of age such that they maintained an upright stance; rats were housed in custom-made cages. Nonamputated rats, also 1 month of age, were kept in regular cages and served as a control group. The lumbar IVDs were harvested from rats in 2 groups, at 5 or 9 months following amputation. Senescence-associated-β-galactosidase- positive staining was used to detect cell senescence. p16INK4a and p27KIP were assessed by immunohistochemistry analysis. Total RNA isolated from these samples was used to measure the gene expression of p16INK4a, RB, cyclin D1, CDK4, PTEN, p27KIP, p19ARF, p21, TERT, and RAGE by real-time polymerase chain reaction assay. Results: The highest levels of SA-β-GAL activity were detected in 9-month amputated rats. Quantitative immunohistochemical analysis showed that there were highest rates of p16INK4a and p27KIP protein expression in the cartilage endplate and anulus fibrosus of 9-month amputated rats. The mRNA levels of p16INK4a, RB, PTEN, p27KIP, p19ARF, and RAGE were upregulated. The increased cyclin D1 mRNA level was statistically significant only at the ninth month following amputation; CDK4 and TERT mRNA levels were downregulated to a similar extent at both points compared with nonamputated controls. mRNA expression of p21 was significantly downregulated. Conclusion: Accelerated cell senescence was associated with forelimb amputation that causes abnormal loading in rat lumbar IVDs.

AB - Study Design: Several senescence biomarkers were observed to investigate cell senescence in degenerative intervertebral lumbar discs of foreleg-amputated rats. Objective: To determine if cell senescence is accelerated in degenerative intervertebral lumbar disc cells in an upright-rat model. Summary Of Background Data: Cellular senescence was accelerated in human and sand rat degenerative intervertebral disc (IVD) cells. Repeated use of upright posture by rats contributed to degenerative disc changes. No convincing evidence of cell senescence was observed in the lumbar disc of the foreleg amputated rat. Methods: The forelimbs of 20 rats were amputated at 1 month of age such that they maintained an upright stance; rats were housed in custom-made cages. Nonamputated rats, also 1 month of age, were kept in regular cages and served as a control group. The lumbar IVDs were harvested from rats in 2 groups, at 5 or 9 months following amputation. Senescence-associated-β-galactosidase- positive staining was used to detect cell senescence. p16INK4a and p27KIP were assessed by immunohistochemistry analysis. Total RNA isolated from these samples was used to measure the gene expression of p16INK4a, RB, cyclin D1, CDK4, PTEN, p27KIP, p19ARF, p21, TERT, and RAGE by real-time polymerase chain reaction assay. Results: The highest levels of SA-β-GAL activity were detected in 9-month amputated rats. Quantitative immunohistochemical analysis showed that there were highest rates of p16INK4a and p27KIP protein expression in the cartilage endplate and anulus fibrosus of 9-month amputated rats. The mRNA levels of p16INK4a, RB, PTEN, p27KIP, p19ARF, and RAGE were upregulated. The increased cyclin D1 mRNA level was statistically significant only at the ninth month following amputation; CDK4 and TERT mRNA levels were downregulated to a similar extent at both points compared with nonamputated controls. mRNA expression of p21 was significantly downregulated. Conclusion: Accelerated cell senescence was associated with forelimb amputation that causes abnormal loading in rat lumbar IVDs.

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