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Melatonin resists oxidative stress-induced apoptosis in nucleus pulposus cells

He, Ruijun, Cui, Min, Lin, Hui, Zhao, Lei, Wang, Jiayu, Chen, Songfeng, Shao, Zengwu
Life sciences 2018 v.199 pp. 122-130
Western blotting, antioxidant activity, apoptosis, biological clocks, enzymatic reactions, etiology, extracellular matrix, hydrogen peroxide, intervertebral disks, malondialdehyde, melatonin, membrane potential, mitochondria, mitochondrial membrane, oxidation, oxidative stress, pain, proteins, rats, reducing agents, staining, superoxide dismutase
Intervertebral disc degeneration (IVDD) is thought to be the major cause of low back pain (LBP), which is still in lack of effective etiological treatment. Oxidative stress has been demonstrated to participate in the impairment of nucleus pulposus cells (NPCs). As the most important neuroendocrine hormone in biological clock regulation, melatonin (MLT) is also featured by good antioxidant effect. In this study, we investigated the effect and mechanisms of melatonin on oxidative stress-induced damage in rat NPCs.Cytotoxicity of H2O2 and protecting effect of melatonin were analyzed with Cell Counting kit-8 (CCK-8). Cell apoptosis rate was detected by Annexin V-FITC/PI staining. DCFH-DA probe was used for the reactive oxygen species (ROS) detection. The mitochondrial membrane potential (MMP) changes were analyzed with JC-1 probe. Intracellular oxidation product and reductants were measured through enzymatic reactions. Extracellular matrix (ECM) and apoptosis associated proteins were analyzed with Western blot assays.Melatonin preserved cell viability of NPCs under oxidative stress. The apoptosis rate, ROS level and malonaldehyde (MDA) declined with melatonin. MLT/H2O2 group showed higher activities of GSH and SOD. The fall of MMP receded and the expression of ECM protein increased with treatment of melatonin. The mitochondrial pathway of apoptosis was inhibited by melatonin.Melatonin alleviated the oxidative stress-induced apoptosis of NPCs. Melatonin could be a promising alternative in treatment of IVDD.