Chinese Journal of Tissue Engineering Research ›› 2019, Vol. 23 ›› Issue (26): 4115-4120.doi: 10.3969/j.issn.2095-4344.1398

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Resveratrol reduces tumor nuclear factor-α expression in wear-particle-treated macrophages by regulating the level of intracellular oxidative stress

Liu Wenhua, Liang Jinfeng, Deng Shaojie
  

  1. Department of Bone and Joint Diseases, Shenzhen Baoan District People's Hospital, Shenzhen 518101, Guangdong Province, China
  • Received:2019-05-04
  • Contact: Liang Jinfeng, Master, Associate chief physician, Department of Bone and Joint Diseases, Shenzhen Baoan District People's Hospital, Shenzhen 518101, Guangdong Province, China
  • About author:Liu Wenhua, Master, Associate chief physician, Department of Bone and Joint Diseases, Shenzhen Baoan District People's Hospital, Shenzhen 518101, Guangdong Province, China

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Abstract:

BACKGROUND: Numerous in vitro cell and animal experiments have shown that resveratrol exhibits anti-oxidative stress, anti-inflammatory, anti-viral, anti-tumor and anti-aging activities and it also has bone-protecting effect.
OBJECTIVE: To investigate the regulatory effect of resveratrol on oxidative stress and inflammatory response of RAW264.7 macrophages induced by titanium wear-particles.
METHODS: The RAW264.7 macrophages were treated with different doses of resveratrol (0, 10, 20, 40, 80, 160 μmol/L) for 24 hours. Cell viability was detected using WST-1 method. Resveratrol doses that did not produce significant change in cell viability were used in the subsequent experiments. RAW264.7 cells were divided into five groups. In the control group, cells were routinely cultured. In the titanium group, RAW264.7 macrophages were treated with titanium particles for 12 hours. In the low-, medium-, and high-dose resveratrol groups, RAW264.7 macrophages were pre-treated with 10, 20, 40 μmol/L resveratrol for 4 hours, and then they were treated with titanium particles for 12 hours. Thereafter, cells and cell supernatant were collected for measurement of intracellular reactive oxygen species generation using flow cytometry. Total RNA was extracted from cells for measuring mRNA level of anti-oxidative enzymes (Cu/Zn superoxide dismutase, Mn superoxide dismutase, glutathione peroxidase, glutathione reductase and catalase) and tumor necrosis factor-α. Cell supernatant was used for measuring the level of TNF-α using ELISA.
RESULTS AND CONCLUSIONS: Treatment with resveratrol (10, 20, and 40 μmol/L) did not affect the viability of RAW264.7 macrophages. Therefore, these doses (10, 20, and 40 μmol/L) were used in the following experiments. Intracellular reactive oxygen species generation in the titanium group was significantly increased compared with the control group (P < 0.05). Resveratrol decreased reactive oxygen species generation in a dose-dependent manner compared with the titanium group (P < 0.05). Compared with the control group, Mn superoxide dismutase and glutathione peroxidase mRNA levels significantly decreased (P < 0.05), glutathione reductase and catalase mRNA levels significantly increased (P < 0.05) in the titanium group. Compared with the titanium group, Mn superoxide dismutase and Cu/Zn superoxide dismutase mRNA levels significantly increased in the medium- and high-dose resveratrol groups (P < 0.05), and catalase mRNA level significantly increased in the low-, medium-, and high-dose resveratrol groups (P < 0.05). The mRNA level and secretion of tumor necrosis factor-α in the titanium group were significantly increased than in the control group (P < 0.05). Compared with the titanium group, low-, medium-, and high-dose resveratrol decreased the mRNA level and secretion of tumor necrosis factor-α (P < 0.05). These results indicate that resveratrol can effectively enhance the transcription of antioxidant stress-related enzymes in macrophages stimulated by titanium particles, reduce the level of intracellular oxidative stress, and reduce the release of inflammatory factor tumor necrosis factor-α.

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