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Evaluation of apocynin in vitro on high glucose-induced oxidative stress on tenocytes
T Kurosawa1, Y Mifune1, A Inui1
1Department of Orthopaedic Surgery, Kobe University Graduate School of Medicine, Kobe, Japan.
Aims:
The purpose of this study was to evaluate the in vitro effects of apocynin, an inhibitor of nicotinamide adenine dinucleotide phosphate oxidase (NOX) and a downregulator of intracellular reactive oxygen species (ROS), on high glucose-induced oxidative stress on tenocytes.
Methods:
Tenocytes from normal Sprague-Dawley rats were cultured in both control and high-glucose conditions. Apocynin was added at cell seeding, dividing the tenocytes into four groups: the control group; regular glucose with apocynin (RG apo+); high glucose with apocynin (HG apo+); and high glucose without apocynin (HG apo-). Reactive oxygen species production, cell proliferation, apoptosis and messenger RNA (mRNA) expression of NOX1 and 4, and interleukin-6 (IL-6) were determined in vitro.
Results:
Expression of NOX1, NOX4, and IL-6 mRNA in the HG groups was significantly higher compared with that in the RG groups, and NOX1, NOX4, and IL-6 mRNA expression in the HG apo+ group was significantly lower compared with that in the HG apo- group. Cell proliferation in the RG apo+ group was significantly higher than in the control group and was also significantly higher in the HG apo+ group than in the HG apo- group. Both the ROS accumulation and the amounts of apoptotic cells in the HG groups were greater than those in the RG groups and were significantly less in the HG apo+ group than in the HG apo- group.
Conclusion:
Apocynin reduced ROS production and cell death via NOX inhibition in high-glucose conditions. Apocynin is therefore a potential prodrug in the treatment of diabetic tendinopathy.Cite this article: Bone Joint Res 2020;9(1):23-28.
Insights
Apocynin, a nicotinamide adenine dinucleotide phosphate oxidase (NOX) inhibitor, reduced reactive oxygen species (ROS) and cell death in high glucose conditions. This suggests apocynin may treat diabetic tendinopathy.
Area of Science:
- Biochemistry
- Cell Biology
- Pharmacology
Background:
- High glucose conditions induce oxidative stress in tenocytes.
- Nicotinamide adenine dinucleotide phosphate oxidase (NOX) enzymes contribute to reactive oxygen species (ROS) production.
- Diabetic tendinopathy is a complication associated with high glucose levels.
Purpose of the Study:
- To investigate the in vitro effects of apocynin on high glucose-induced oxidative stress in tenocytes.
- To evaluate apocynin's impact on ROS production, cell proliferation, and apoptosis.
- To assess apocynin's influence on NOX1, NOX4, and interleukin-6 (IL-6) mRNA expression.
Main Methods:
- Tenocytes were cultured under regular and high-glucose conditions.
- Apocynin was administered to assess its effects on various cellular parameters.
- Measurements included ROS production, cell proliferation, apoptosis, and mRNA expression of NOX1, NOX4, and IL-6.
Main Results:
- High glucose increased NOX1, NOX4, and IL-6 mRNA expression, ROS accumulation, and apoptosis.
- Apocynin treatment significantly reduced NOX1, NOX4, and IL-6 mRNA levels in high-glucose conditions.
- Apocynin treatment also decreased ROS production and apoptosis while enhancing cell proliferation.
Conclusions:
- Apocynin effectively mitigates high glucose-induced oxidative stress and cell death in tenocytes by inhibiting NOX enzymes.
- Apocynin demonstrates potential as a therapeutic agent for diabetic tendinopathy.
- Further research into apocynin as a prodrug for tendinopathies is warranted.
