| Objective: To investigated the role of HO-1 in IDD by assessing the effects of HO-1 overexpression on IL-1?-induced apoptosis in NPCs.Methods: 16 cases of lumbar disc herniation and 6 cases of lumbar fracture were treated with discectomy,paraffin section was used to fix tissue samples,and enzyme digestion was used to extract primary cell samples.The expression of HO-1 and autophagy level in lumbar disc herniation samples and fracture samples were analyzed by Western blot.The overexpression of HO-1 adenovirus and empty adenovirus were constructed to verify the overexpression effect and regulate the expression level of HO-1.After IL-1 ? was used to construct cell inflammation model and 3-MA or CQ was used to block autophagy,the expression levels of related proteins and complex were detected by immunoprecipitation and Western blot.Furthermore,autophagy and apoptosis were detected by immunofluorescence,transmission electron microscope and flow cytometry.Results: Immunohistochemical staining showed HO-1 expression tobe lower in NPCs from IDD patients than from patients with LVF.Western blot analysis showed HO-1 and LC3-II/I levels to be lower in NP tissues from IDD patients than from LVF patients,suggesting suppression of autophagy in degenerative intervertebral disc.Consistent with that idea,autophagy was increased in HO-1-overexpressing NPCs while IL-1?-induced apoptosis was reduced.These effects were reversed by treatment with the early autophagy inhibitor 3-MA,which suggests HO-1-induced autophagy suppresses IL-1?-induced apoptosis in NPCs.HO-1overexpression promoted autophagy by increasing levels of Beclin-1/PI3KC3 complex.Phospho-P65 levels were lower in HO-1-overexpressing NPCs,suggesting inhibition of NF-?B-mediated apoptosis.Conclusion: HO-1 promotes autophagy by enhancing formation of Beclin-1/PI3KC3 complex and suppresses IL-1?-induced apoptosis by inhibiting NF-?B.We suggest that HO-1 is a potential therapeutic target to alleviate IDD. |
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