Effects of nitric oxide and nitric oxide synthase inhibitors on mitochondrial function of nucleus pulposus cells

doi:10.3969/j.issn.2095-4344.2016.42.007

Abstract

Abstract:

BACKGROUND: Nitric oxide can interfere with the function of mitochondria, and accelerate the intervertebral disc damage and degeneration by interfering with the release of inflammatory cytokines. Nitric oxide is an important inflammatory cell medium leading to degeneration of intervertebral disc induced by pressure and other external factors.
OBJECTIVE: To investigate the regulatory effect of nitric oxide and nitric oxide synthase inhibitor niacinamide on mitochondrial function and its association with biological behavior of rabbit nucleus pulposus.
METHODS: Cultured nucleus pulposus cells of rabbit lumbar intervertebral disc were randomly divided into six groups: normal blank control group, 10 μmol/L sodium nitroprusside group, 100 μmol/L sodium nitroprusside group, 200 μmol/L sodium nitroprusside group, 0.05 g/L nicotinamide group (100 μmol/L sodium nitroprusside+0.05g/L nicotinamide), and 0.5 g/L nicotinamide group (100 μmol/L sodium nitroprusside and 0.5 g/L nicotinamide). Different doses of nitric oxide donor sodium nitroprusside and nicotinamide were added in the medium of each group. Three days after intervention, cell proliferation activity, intracellular ATP concentration, cell nitric oxide synthase activity, cellular reactive oxygen species level, and mitochondrial membrane potential were detected respectively.
RESULTS AND CONCLUSION: (1) After 3 days of rabbit nucleus pulposus cells intervened by different concentrations of sodium nitroprusside, intracellular nitric oxide synthase content increased with sodium nitroprusside volume increase, and ATP concentration decreased along with sodium nitroprusside volume increase; there were significantly differences between the normal control group and sodium nitroprusside groups (P < 0.01). (2) Reactive oxygen species could be increased in the sodium nitroprusside group. Niacinamide groups indicated a dose-dependent manner to improve the increase of cellular reactive oxygen species levels with sodium nitroprusside intervention (P < 0.01). (3) In the sodium nitroprusside groups, nucleus pulposus cell membrane potential decreased. In the niacinamide groups, sodium nitroprusside- induced decline in mitochondrial membrane potential was reduced (P < 0.01). (4) Niacinamide groups also indicated a dose-dependent manner to improve the proliferative activity of nucleus pulposus cells as compared with sodium nitroprusside groups (P < 0.01). Significant differences were determined between the two groups (P < 0.01). (5) Results suggest that the excess nitric oxide can damage mitochondrial metabolic function of rabbit nucleus pulposus cells and cause cell energy metabolism. Niacinamide can reverse these damages by inhibiting nitric oxide synthesis, thereby contributing to the prevention against intervertebral disc degeneration.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Mitochondria, Nitric Oxide, Nitric Oxide Synthase, Tissue Engineering

CLC Number:

R318

Zhou Jian-guo1, Yang Cao2, Xiong Li-ming2. Effects of nitric oxide and nitric oxide synthase inhibitors on mitochondrial function of nucleus pulposus cells[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(42): 6278-6283.

Figures/Tables 3

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