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Repression of P66Shc expression by SIRT1 contributes to the prevention of hyperglycemia-induced endothelial
Shuang Zhou1, Hou-Zao Chen, Yan-Zhen Wan
1National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100005, PR China.
Rationale:
Inactivation of the p66Shc adaptor protein confers resistance to oxidative stress and protects mice from aging-associated vascular diseases. However, there is limited information about the negative regulating mechanisms of p66Shc expression in the vascular system.
Objective:
In this study, we investigated the role of SIRT1, a class III histone deacetylase, in the regulation of p66Shc expression and hyperglycemia-induced endothelial dysfunction.
Methods And Results:
Expressions of p66Shc gene transcript and protein were significantly increased by different kinds of class III histone deacetylase (sirtuin) inhibitors in human umbilical vein endothelial cells and 293A cells. Adenoviral overexpression of SIRT1 inhibited high-glucose-induced p66Shc upregulation in human umbilical vein endothelial cells. Knockdown of SIRT1 increased p66Shc expression and also increased the expression levels of plasminogen activator inhibitor-1 expression, but decreased manganese superoxide dismutase expression in high-glucose conditions. However, knockdown of p66Shc significantly reversed the effects of SIRT1 knockdown. In addition, p66Shc overexpression significantly decreased manganese superoxide dismutase expression and increased plasminogen activator inhibitor-1 expression in high-glucose conditions, which were recovered by SIRT1 overexpression. Moreover, compared to streptozotocin-induced wild-type diabetic mice, endothelium-specific SIRT1 transgenic diabetic mice had decreased p66Shc expression at both the mRNA and the protein levels, improved endothelial function, and reduced accumulation of nitrotyrosine and 8-OHdG (markers of oxidative stress). We further found that SIRT1 was able to bind to the p66Shc promoter (-508 bp to -250 bp), resulting in a decrease in the acetylation of histone H3 bound to the p66Shc promoter region.
Conclusion:
Our findings indicate that repression of p66Shc expression by SIRT1 contributes to the protection of hyperglycemia-induced endothelial dysfunction.
Insights
SIRT1 represses p66Shc expression, protecting against high-glucose-induced endothelial dysfunction. This finding reveals a key mechanism in vascular aging and diabetes complications.
Area of Science:
- Vascular Biology
- Epigenetics
- Metabolic Disease
Background:
- p66Shc adaptor protein inactivation confers oxidative stress resistance and protects against aging-associated vascular diseases.
- Limited information exists on negative regulatory mechanisms of p66Shc expression in the vascular system.
Purpose of the Study:
- Investigate the role of SIRT1 (a class III histone deacetylase) in regulating p66Shc expression.
- Determine SIRT1's role in hyperglycemia-induced endothelial dysfunction.
Main Methods:
- Utilized human umbilical vein endothelial cells and 293A cells treated with sirtuin inhibitors.
- Employed adenoviral overexpression and knockdown techniques for SIRT1 and p66Shc.
- Analyzed gene and protein expression, endothelial function, and oxidative stress markers in diabetic mouse models.
Main Results:
- Sirtuin inhibitors increased p66Shc expression; SIRT1 overexpression inhibited high-glucose-induced p66Shc upregulation.
- SIRT1 knockdown increased p66Shc, plasminogen activator inhibitor-1, and decreased manganese superoxide dismutase; p66Shc knockdown reversed these effects.
- Endothelium-specific SIRT1 transgenic diabetic mice showed decreased p66Shc, improved endothelial function, and reduced oxidative stress markers compared to wild-type diabetic mice.
- SIRT1 binds to the p66Shc promoter, decreasing histone H3 acetylation.
Conclusions:
- SIRT1 represses p66Shc expression, contributing to the protection against hyperglycemia-induced endothelial dysfunction.
- This study elucidates a novel epigenetic mechanism involving SIRT1 and p66Shc in vascular health under diabetic conditions.
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