The Study Of Aspirin Therapeutic Effect On Intervertebral Disc Degeneration And Mechanism By Regulation Of Oxidative Stress

Objective:Intervertebral disc degeneration(IDD)is one of the important causes of low back pain.Studies have shown that the pathological changes of IDD are closely related to reactive oxygen species increase,inflammation highly express and matrix loss.Aspirin is one of the classic non-steroidal anti-inflammatory drugs,which can effectively alleviate all kinds of pain and inflammation in clinical applications.However,whether Aspirin can improve IDD process by mitigating oxidative stress is still unclear.The aim of our study was to investigate the effects of Aspirin on the regulation of oxidative stress in relieving IDD and to propose new treatment strategy.Method:The nucleus pulposus of the caudal vertebrae of Sprague Dawley rats was isolated under sterile conditions.Lipopolysaccharide(LPS,1 ?g/mL)induced the oxidative stress in nucleus pulposus cells(NPCs),then Aspirin(5 ?g/mL or 25 ?g/mL)was added to the culture medium.The expression levels of reactive oxygen species(ROS)and inflammatory factors were measured,and the degeneration degree of NPCs was observed by real-time PCR and cell immunofluorescence staining.In the vivo,degeneration was induced by a rat caudal disc puncture model,and local intervention with Aspirin(10 ?g/mL or 100?g/mL).One week later,the rats were sacrificed.The imaging and histomorphology of the caudal intervertebral disc were taken and analyzed by immunohistochemical staining.In the mechanism research,Compound C which is the specific inhibitor of AMP-activated protein kinase(AMPK)was used to block the AMPK signaling pathway and observe the effect on the protective effect of Aspirin.Western-blot was used to clarify the specific mechanism of Aspirin to alleviate IDD.Result:In the vitro,results showed that LPS could significantly induce oxidative stress and stimulate NPCs to secrete a large amount of ROS and inflammatory factors.NPCs were lost of Collagen-2,Aggrecan and high expression of MMP-3 and MMP-13.Finally,NPCs were induced degeneration.The Aspirin can effectively inhibit the expression of oxidative stress and related inflammatory factors,thereby alleviating the development of degeneration.In the vivo,results were show that Aspirin can reduce the water loss of nucleus pulposus tissue and improve MRI signal,and can relieve the decrease of the high of intervertebral disc.In the mechanism research,the protective effect of Aspirin was observed by using AMPK-specific inhibitor Compound C,and it was further confirmed that Aspirin could protect NPCs through the AMPK-Acetyl-CoA carboxylase(ACC)signaling pathway.Conclusion:Our results clarify that the important role of oxidative stress in IDD.Demonstrating that Aspirin can act as a drug of relieving oxidative stress to improve IDD process.Meanwhile,AMPK plays an important role in relieving disc degeneration and can also be used as an important intervention target to treat other oxidative stress-related diseases.