Caspase 3 silencing inhibits biomechanical overload-induced intervertebral disk degeneration

Am J Pathol. 2014 Mar;184(3):753-64. doi: 10.1016/j.ajpath.2013.11.010. Epub 2014 Jan 3.

Abstract

Intervertebral disk (IVD) degeneration causes debilitating low back pain in much of the worldwide population. No efficient treatment exists because of an unclear pathogenesis. One characteristic event early in such degeneration is the apoptosis of nucleus pulposus (NP) cells embedded in IVDs. Excessive biomechanical loading may also be a major etiology of IVD degeneration. The present study used in vitro and in vivo models of compressive loading to elucidate the underlying mechanism of IVD degeneration. In addition, we investigated whether the inhibition of apoptosis is a potential clinical therapeutic strategy for the treatment of IVD degeneration induced by biomechanical stress. A TUNEL assay showed that NP cell-agarose three-dimensional composite cultures subjected to uniaxial, unconfined, static, compressive loading exhibited a time-dependent increase in apoptosis. Western blot analysis revealed the up-regulation of several extracellular matrix-degrading enzymes and down-regulation of tissue inhibitor of metalloproteinase 1. These responses to compressive loading were all significantly inhibited by caspase 3 siRNA. In the in vivo model of compressive loading-induced IVD degeneration, a single local injection of caspase 3 siRNA significantly inhibited IVD degeneration by magnetic resonance imaging, histological findings, IHC, and TUNEL assay. The present study suggests that caspase 3 siRNA attenuates overload-induced IVD degeneration by inhibiting NP cell apoptosis and the expression of matrix-degrading enzymes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis*
  • Biomechanical Phenomena
  • Caspase 3 / genetics
  • Caspase 3 / metabolism*
  • Disease Models, Animal
  • Down-Regulation
  • Extracellular Matrix / metabolism
  • Gene Expression Regulation, Enzymologic*
  • Gene Silencing
  • Humans
  • Intervertebral Disc / metabolism
  • Intervertebral Disc / pathology
  • Intervertebral Disc Degeneration / etiology
  • Intervertebral Disc Degeneration / pathology*
  • Intervertebral Disc Degeneration / physiopathology*
  • Intervertebral Disc Degeneration / therapy
  • Male
  • Models, Biological
  • RNA, Small Interfering / administration & dosage
  • RNA, Small Interfering / metabolism
  • Rabbits
  • Signal Transduction
  • Up-Regulation
  • Weight-Bearing

Substances

  • RNA, Small Interfering
  • Caspase 3