Abstract
Aim:
To observe the effect of Rhein lysinate (RHL) on cellular senescence of human umbilical vascular endothelial cells (HUVECs) and elucidate its action mechanism.
Methods:
Cell viability was determined using MTT assay. The expression levels of Sirt1 mRNA and protein were measured by RT-PCR and Western blot, respectively. Senescence associated (SA)-β-galactosidase activity was detected to evaluate cell senescence. Apoptosis and cell cycle progression were determined using flow cytometry.
Results:
Treatment with RHL (10 μmol/L) for 48 h significantly increased the proliferation of HUVECs. In contrast, treatment with H2O2 (25, 50 and 100 μmol/L) for 6 d dose-dependently increased β-galactosidase positive cells. Spontaneous cell senescence appeared as the cell passage increased. Pre-treatment with RHL (10 μmol/L) reversed H2O2 or increased cell passage-induced cell senescence. H2O2 (100 μmol/L) significantly arrested HUVECs at G1 phase (73.8% vs 64.6% in the vehicle group), which was blocked by RHL (10 μmol/L). RHL (5 and 10 μmol/L) enhanced both mRNA transcription and protein expression of Sirt1. H2O2 (100 μmol/L) significantly decreased Sirt1 expression, and induced up-regulation of p53 acetylation and p16INK4a, which were blocked by pre-treatment with RHL (10 μmol/L). Interference with siRNA for Sirt1 abolished the effect of RHL. H2O2 (100 μmol/L) did not induce HUVEC apoptosis. The expression of apoptosis-associated proteins, such as p53, p21, Bcl-2, and Bax, did not significantly change in the presence of H2O2 (100 μmol/L) or RHL (10 μmol/L).
Conclusion:
RHL protected HUVECs against cellular senescence induced by H2O2, via up-regulation of Sirt1 expression and down-regulation of the expression of acetyl-p53 and p16INK4a.
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This study was supported by a grant from the National Natural Science Foundation of China (No 81001439).
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Lin, Yj., Zhen, Yz., Wei, J. et al. Effects of Rhein Lysinate on H2O2-induced cellular senescence of human umbilical vascular endothelial cells. Acta Pharmacol Sin 32, 1246–1252 (2011). https://doi.org/10.1038/aps.2011.101
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DOI: https://doi.org/10.1038/aps.2011.101
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