A steroid like phytochemical Antcin M is an anti-aging reagent that eliminates hyperglycemia-accelerated premature senescence in dermal fibroblasts by direct activation of Nrf2 and SIRT-1

// Kumar K.J. Senthil 1 , Vani M. Gokila 1,2 , Jeng-Leun Mau 2,3 , Chin-Chung Lin 4 , Fang-Hua Chu 5 , Chia-Cheng Wei 6 , Vivian Hsiu-Chuan Liao 6 and Sheng-Yang Wang 1,2,7 1 Department of Forestry, National Chung Hsing University, Taichung, Taiwan 2 National Chung Hsing University/University of California at Davis, Plant and Food Biotechnology Center, National Chung Hsing University, Taichung, Taiwan 3 Department of Food Science and Biotechnology, National Chung Hsing University, Taichung, Taiwan 4 Taiwan Leader Biotech Company, Taipei, Taiwan 5 School of Forestry and Resource Conservation, National Taiwan University, Taipei, Taiwan 6 Department of Bioenvironmental Systems Engineering, National Taiwan University, Taipei, Taiwan 7 Agricultural Biotechnology Research Center, Academia Sinica, Taipei, Taiwan Correspondence to: Sheng-Yang Wang, email: // Keywords : Antcin M, antrodia salmonea , hyperglycemia, stress-induced premature senescence, SIRT-1, Gerotarget Received : February 18, 2016 Accepted : July 27, 2016 Published : August 11, 2016 Abstract The present study revealed the anti-aging properties of antcin M (ANM) and elucidated the molecular mechanism underlying the effects. We found that exposure of human normal dermal fibroblasts (HNDFs) to high-glucose (HG, 30 mM) for 3 days, accelerated G 0 /G 1 phase arrest and senescence. Indeed, co-treatment with ANM (10 µM) significantly attenuated HG-induced growth arrest and promoted cell proliferation. Further molecular analysis revealed that ANM blocked the HG-induced reduction in G 1 -S transition regulatory proteins such as cyclin D, cyclin E, CDK4, CDK6, CDK2 and protein retinoblastoma (pRb). In addition, treatment with ANM eliminated HG-induced reactive oxygen species (ROS) through the induction of anti-oxidant genes, HO-1 and NQO-1 via transcriptional activation of Nrf2. Moreover, treatment with ANM abolished HG-induced SIPS as evidenced by reduced senescence-associated β-galactosidase (SA-β-gal) activity. This effect was further confirmed by reduction in senescence-associated marker proteins including, p21 CIP1 , p16 INK4A , and p53/FoxO1 acetylation. Also, the HG-induced decline in aging-related marker protein SMP30 was rescued by ANM. Furthermore, treatment with ANM increased SIRT-1 expression, and prevented SIRT-1 depletion. This protection was consistent with inhibition of SIRT-1 phosphorylation at Ser47 followed by blocking its upstream kinases, p38 MAPK and JNK/SAPK. Further analysis revealed that ANM partially protected HG-induced senescence in SIRT-1 silenced cells. A similar effect was also observed in Nrf2 silenced cells. However, a complete loss of protection was observed in both Nrf2 and SIRT-1 knockdown cells suggesting that both induction of Nrf2-mediated anti-oxidant defense and SIRT-1-mediated deacetylation activity contribute to the anti-aging properties of ANM in vitro . Result of in vivo studies shows that ANM-treated C. elegens exhibits an increased survival rate during HG-induced oxidative stress insult. Furthermore, ANM significantly extended the life span of C. elegans . Taken together, our results suggest the potential application of ANM in age-related diseases or as a preventive reagent against aging process.