Antcin M as an anti-aging agent for eliminating hyperglycemia-accelerated premature senescence in dermal fibroblasts by direct activation of Nrf2 and SIRT-1 characterized in that the survival rate of ANM-treated Caenorhabditis elegans increases during the HG-induced oxidative stress injury period
The present invention discloses the molecular mechanism of anti-aging properties of antcin M (ANM). This invention has found that human normal dermal fibroblast (HNDF) accelerates the arrest and aging in G0/G1 phase after exposure to high-glucose (HG, 30 mM) for 3 days. Experiments confirmed that co-treatment with ANM (10[mu]M) may significantly reduce HG-induced growth arrest and promoted cell proliferation. Further analysis revealed that ANM can inhibit HG-induced G1-S transitional protein reduction, such as cyclin D, cyclin E, CDK4, CDK6, CDK2 and protein retinoblastoma (pRb). In addition, under the ANM treatment, transcriptional activation of Nrf2 can induce the antioxidant genes HO-1 and NQO-1 and eliminates the HG-induced reactive oxygen species (ROS). It was demonstrated that ANM treatment can eliminate stress-induced premature senescence (SIPS) in the presence of HG by reducing the activity of senescence-associated [beta]-galactosidase (SA-[beta]-gal). This activity can be demonstrated by reducing the acetylated P21CIP1, p16INK4A and p53/FoxO1 of marker proteins associated with aging. In addition, cells administrated with ANM may enhance HG-induced expression of the aging-associated marker protein SMP30 and increased the expression of SIRT-1 as well as prevented the reduction of SIRT-1. This protection was confirmed by the consistency of inhibiting the phosphorylation of SIRT-1 at Ser47 by blocking the upstream kinases of p38 MAPK and JNK/SAPK. Further analysis showed that ANM protects some SIRT-1 silenced cells from HG-induced aging, and similar effect can be observed in Nrf2 silenced cells. However, a complete loss of this protective effect was observed in cells both Nrf2 and SIRT-1 are knockdown and eliminated, indicating that the antioxidant defense of Nrf2 and the deacetylation activity of the deacetylated SIRT-1 mediators contribute to the anti-aging activity of ANM in vitro. The results of the in vivo studies show that the survival rate of ANM-treated
