Mitochondrial Pathway Is Involved in Advanced Glycation End Products-Induced Apoptosis of Rabbit Annulus Fibrosus Cells

Study Design. Experimental study. Objective. The purposes of this study were to evaluate whether advanced glycation end-products (AGEs) induce annulus fibrosus (AF) cell apoptosis and further to explore the mechanism by which this process occurs. Summary of Background Data. Recent studies revealed that AGEs accumulation is considered an important factor in diabetic intervertebral disc (IVD) degeneration. However, the effect of AGEs on intervertebral disc remains unclear. Methods. AF cells were treated with various concentrations of AGEs for 3 days. Cell viability and cell proliferation were measured by Cell Counting Kit-8 (CCK-8) and 5-ethynyl-2′-deoxyuridine (EdU) incorporation assays, respectively. Cell apoptosis was examined by Annexin V/PI apoptosis detection kit and Hoechst 33342. The expression of apoptosis-related proteins, including Bax, Bcl-2, cytochrome c, caspase-3, and caspase-9, was detected by western blotting. In addition, Bax and Bcl-2 mRNA expression levels were detected by real-time PCR (RT-PCR). Mitochondrial membrane potential (MMP) and intracellular reactive oxygen species (ROS) production of AF cell were examined by 5,5′,6,6′ -Tetrachloro-1,1′,3,3′- tetraethyl-imidacarbocyanine iodide (JC-1) staining and 2′,7′-Dichlorodihydrofluorescein diacetate (DCFH-DA) fluorescent probes, respectively. Results. Our results indicated that AGEs had inhibitory effects on AF cell proliferation and induced AF cell apoptosis. The molecular data showed that AGEs significantly up-regulated Bax expression and inhibited Bcl-2 expression. In addition, AGEs increased the release of cytochrome c into the cytosol and enhanced caspase-9 and caspase-3 activation. Moreover, treatment with AGEs resulted in a decrease in MMP and the accumulation of intracellular ROS in AF cells. The antioxidant N-acetyl-L-cysteine (NAC) significantly reversed AGE-induced MMP decrease and AF cell apoptosis. Conclusion. These results suggested that AGEs induce rabbit AF cell apoptosis and mitochondrial pathway may be involved in AGEs-mediated cell apoptosis, which may provide a theoretical basis for diabetic IVD degeneration. Level of Evidence: N/A

[1]  W. Kuo,et al.  Pkcδ Activation is Involved in ROS-Mediated Mitochondrial Dysfunction and Apoptosis in Cardiomyocytes Exposed to Advanced Glycation End Products (Ages) , 2018, Aging and disease.

[2]  K. Horiuchi,et al.  Potential Involvement of the IL-6/JAK/STAT3 Pathway in the Pathogenesis of Intervertebral Disc Degeneration , 2017, Spine.

[3]  Yu-fei Chen,et al.  Role of miR‐155 in the regulation of MMP‐16 expression in intervertebral disc degeneration , 2017, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[4]  S. Kuang,et al.  Myricitrin Alleviates Oxidative Stress-induced Inflammation and Apoptosis and Protects Mice against Diabetic Cardiomyopathy , 2017, Scientific Reports.

[5]  Xigao Cheng,et al.  Autophagy Protects Advanced Glycation End Product-Induced Apoptosis and Expression of MMP-3 and MMP-13 in Rat Chondrocytes , 2017, BioMed research international.

[6]  Lin Li,et al.  Protective effects of physiological testosterone on advanced glycation end product-induced injury in human endothelial cells , 2017, Molecular medicine reports.

[7]  Li Xu,et al.  Inhibition of autophagy increased AGE/ROS-mediated apoptosis in mesangial cells , 2016, Cell Death & Disease.

[8]  Jianru Wang,et al.  Resistin Promotes Intervertebral Disc Degeneration by Upregulation of ADAMTS-5 Through p38 MAPK Signaling Pathway , 2016, Spine.

[9]  Chi Ma,et al.  Pioglitazone inhibits advanced glycation end product-induced matrix metalloproteinases and apoptosis by suppressing the activation of MAPK and NF-κB , 2016, Apoptosis.

[10]  Xianzhe Liu,et al.  Long noncoding RNA MALAT1 as a potential therapeutic target in osteosarcoma , 2016, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[11]  R. Agius,et al.  Bone mineral density and intervertebral disc height in type 2 diabetes. , 2016, Journal of diabetes and its complications.

[12]  Caiguo Zhang,et al.  Molecular mechanisms of cell death in intervertebral disc degeneration (Review) , 2016, International journal of molecular medicine.

[13]  Xianzhe Liu,et al.  Establishment and characterization of a novel osteosarcoma cell line: CHOS , 2016, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[14]  T. Kubo,et al.  Hepatocyte growth factor/c‐met promotes proliferation, suppresses apoptosis, and improves matrix metabolism in rabbit nucleus pulposus cells in vitro , 2016, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[15]  L. Xiong,et al.  Ropivacaine- and bupivacaine-induced death of rabbit annulus fibrosus cells in vitro: involvement of the mitochondrial apoptotic pathway. , 2015, Osteoarthritis and cartilage.

[16]  Q. Su,et al.  Sesamin Ameliorates Advanced Glycation End Products-Induced Pancreatic β-Cell Dysfunction and Apoptosis , 2015, Nutrients.

[17]  Stephanie L. Miller,et al.  Alterations in intervertebral disc composition, matrix homeostasis and biomechanical behavior in the UCD‐T2DM rat model of type 2 diabetes , 2015, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[18]  Zhixin Chen,et al.  Advanced glycation end products‐induced chondrocyte apoptosis through mitochondrial dysfunction in cultured rabbit chondrocyte , 2015, Fundamental & Clinical Pharmacology.

[19]  Bo Li,et al.  The Responses of Autophagy and Apoptosis to Oxidative Stress in Nucleus Pulposus Cells: Implications for Disc Degeneration , 2014, Cellular Physiology and Biochemistry.

[20]  C. Niu,et al.  Advanced glycation end products in degenerative nucleus pulposus with diabetes , 2014, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[21]  M. Chordia,et al.  Novel Treatment of Neuroinflammation Against Low Back Pain by Soluble Fullerol Nanoparticles , 2013, Spine.

[22]  James D. Kang,et al.  Mitochondrial‐derived reactive oxygen species (ROS) play a causal role in aging‐related intervertebral disc degeneration , 2013, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[23]  Xiangyang Wang,et al.  Apoptosis, senescence, and autophagy in rat nucleus pulposus cells: Implications for diabetic intervertebral disc degeneration , 2013, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[24]  Fan Ding,et al.  Cell death in intervertebral disc degeneration , 2013, Apoptosis.

[25]  Hao Peng,et al.  The correlation between microvessel pathological changes of the endplate and degeneration of the intervertebral disc in diabetic rats , 2012, Experimental and therapeutic medicine.

[26]  Fan Ding,et al.  Role of mitochondrial pathway in compression-induced apoptosis of nucleus pulposus cells , 2012, Apoptosis.

[27]  G. Striker,et al.  AGE restriction in diabetes mellitus: a paradigm shift , 2011, Nature Reviews Endocrinology.

[28]  Tetsuro Ohba,et al.  Mechanism of Signal Transduction in Tumor Necrosis Factor-Like Weak Inducer of Apoptosis-Induced Matrix Degradation by MMP-3 Upregulation in Disc Tissues , 2008, Spine.

[29]  Su-Yen Goh,et al.  The role of advanced glycation end products in progression and complications of diabetes , 2008 .

[30]  Sten Orrenius,et al.  Mitochondria, oxidative stress and cell death , 2007, Apoptosis.

[31]  L. Dai,et al.  Programmed cell death in intervertebral disc degeneration , 2006, Apoptosis.

[32]  S. Yamagishi,et al.  Advanced glycation end-products downregulating intervertebral disc cell production of proteoglycans in vitro. , 2006, Journal of neurosurgery. Spine.

[33]  Nick Sakellaridis,et al.  The influence of diabetes mellitus on lumbar intervertebral disk herniation. , 2006, Surgical neurology.

[34]  Jaime Uribarri,et al.  Glucose, Advanced Glycation End Products, and Diabetes Complications: What Is New and What Works , 2003 .

[35]  J C Reed,et al.  Mitochondria and apoptosis. , 1998, Science.

[36]  I. Shapiro,et al.  Role of cytokines in intervertebral disc degeneration: pain and disc content , 2014, Nature Reviews Rheumatology.

[37]  Y. Mirovsky,et al.  Diabetes mellitus as a risk factor for the development of lumbar spinal stenosis. , 2010, The Israel Medical Association journal : IMAJ.

[38]  D. Vashishth,et al.  Effects of collagen crosslinking on tissue fragility. , 2008, Clinical biomechanics.

[39]  Jeffrey C Lotz,et al.  Glycation increases human annulus fibrosus stiffness in both experimental measurements and theoretical predictions. , 2006, Journal of biomechanics.

[40]  J. Petrik,et al.  Oxidative stress and cell death , 2003 .