J Korean Med Sci.  2013 Jun;28(6):939-945. 10.3346/jkms.2013.28.6.939.

Resveratrol Has Anabolic Effects on Disc Degeneration in a Rabbit Model

• Kwon YJ

• KMID: 2158045
• DOI: http://doi.org/10.3346/jkms.2013.28.6.939

Abstract

This study was done to evaluate whether injections of resveratrol, a natural compound found in the skin of grapes, had anabolic effects on degenerated intervertebral discs in a rabbit model. Two non-continuous lumbar discs were punctured in rabbits to induce disc degeneration. Four weeks and 6 weeks after puncture, the rabbits were treated by injections with dimethylsulfoxide (DMSO) or resveratrol. At 4, 8, and 16 weeks after initial injection, rabbits were sacrificed and the spine was extracted for magnetic resonance image (MRI), mRNA expression, and histological staining. Resveratrol treatment resulted in stronger signal intensity in T2-weighted images. MRI grade showed significantly lower in the resveratrol group than the DMSO group (P = 0.039). In the resveratrol group, aggrecan gene expression was significantly increased than that in the DMSO group at 16 weeks after injection (P = 0.027). MMP-13 mRNA levels in the resveratrol group were significantly decreased than those in the DMSO group at 8 and 16 weeks (P = 0.006 and P = 0.048, respectively). In hematoxylin and eosin stain, resveratrol-treated discs showed the features of regeneration. Histologic grade revealed improvement in resveratrol-treated discs, compared with DMSO-treated discs (P = 0.024). These anabolic effects on degenerated discs indicate that resveratrol is a promising candidate for treatment of degenerative disc disease.

Keyword

Resveratrol; Model, Animal; Intervertebral Disc Degeneration; Regeneration

MeSH Terms

Aggrecans/genetics/metabolism
Anabolic Agents/*administration & dosage
Animals
Disease Models, Animal
Drug Administration Schedule
Intervertebral Disc Degeneration/*drug therapy/pathology
Magnetic Resonance Imaging
Matrix Metalloproteinase 13/genetics/metabolism
RNA, Messenger/metabolism
Rabbits
Spine/radiography
Stilbenes/*administration & dosage
Aggrecans
Anabolic Agents
RNA, Messenger
Stilbenes
Matrix Metalloproteinase 13

Figure

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  Fig. 1 Percutaneous needle puncture into the rabbit discs under a fluoroscope. Generally, a rabbit has seven lumbar vertebrae. After localization of the exact levels of L2-L3 and L4-L5 (arrows) using the fluoroscope, 18 G angiography needles (radio opaque materials in the figure) are inserted into the discs under a real time monitoring.

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  Fig. 2 Ex vivo analyses of magnetic resonance image (MRI) between the dimethylsulfoxide (DMSO) group and the resveratrol group. Representative T2-weighted sagittal image of lumbar spine demonstrated that resveratrol-treated discs (B) showed higher signal intensity than DMSO-treated discs (A) (arrows; treated discs of L2-L3 and L4-L5). Analysis of MRI data pooled from three time points. MRI grades show significantly lower in the resveratrol group than the DMSO group (*P = 0.039). (C) Data are presented as mean±SD.

• 

  Fig. 3 Gene expression analyses of aggrecan and matrix metalloprotease-13 (MMP-13) between the DMSO group and the resveratrol group. (A) Aggrecan mRNA level in resveratrol-treated discs is significantly increased than that in DMSO-treated discs at 16 weeks post treatment (*P = 0.027). (B) MMP-13 mRNA levels significantly is decreased in resveratrol-treated discs than those in DMSO-treated discs at 8 and 16 weeks (†P = 0.006 and ‡P = 0.048, respectively). Data are presented as mean±SD.

• 

  Fig. 4 Histologic analyses of intervertebral disc tissues treated with DMSO and resveratrol. 40 × (A, B, and C) and 100 × (a, b, and c) power images of hematoxylin and eosin staining. Normal disc has a clear demarcation between the nucleus pulposus (NP) and the annulus fibrosus, and abundant extracellular matrix (ECM) (A and a). The degenerated, DMSO-treated disc loses the characteristics. They show indistinct NP margin and severe fibrosis in ECM (B). High power image reveals many fibroblast-like cells, suggesting degeneration (b). In contrast, the resveratrol-treated disc shows the features of restoration. At higher magnification, large, round, grape-like clusters of chondrocytic cells in the NP of the resveratrol-treated disc (c). Significant lower histological grades are noted in the resveratrol group when compared with the DMSO group (*P = 0.024) (D). Data are presented as mean ± SD.

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