Conclusion
Pfirrmann grade III disc degeneration showed that autophagosomes were formed, which led to the reversible decomposition of degenerative substances. Thus, by analyzing the effect of autophagy on degenerative discs, we showed the possibility of reversing degenerative changes, but only in grades III and lower.
Methods
For the 16 patients included in the present study, we determined the Pfirrmann classifications of degenerative lesions by MRI. Western blot analysis, LC3, LAMP2, and p62 protein expressions were quantified in different degeneration grades of disc nucleus pulposus. Double immunofluorescence staining was used to show co-localization of LC3 and LAMP2, and immunohistochemistry to show LC3 and p62 in the nucleus pulposus.
Results
In the western blot analysis, LC3-II was highly expressed in grade III and decreased progressively from grade IV to V. In addition, LC3-II expression in grade III was significantly higher than in grade II, IV, and V (P < 0.05). LAMP2 expression in grade V was significantly higher than that in grade II, III, and IV (P < 0.05). P62 increased with decreasing autophagy expression up through grade V. In the double staining, the LC3 level was highly expressed in grade III and decreased progressively from grades IV to V, as in the western blot analysis. LAMP2 rose with increasing disc degeneration grade through grade V. In the quantitative analysis of colocalization, grade III is significantly higher than grade II and V (P < 0.05). Immunohistochemical staining showed that LC3 was highly expressed in grade III but weakly expressed in other grades, with few positive areas around the nucleus pulposus. However, p62 increased progressively with increasing disc degeneration grade.