Anti-aging effects of Schisandrin C on H2O2-induced HaCaT

Abstract

Objective　To investigate the effects of Schisandrin C (SchC) on hydrogen peroxide (H2O2) treated human immortalized keratinocyte cells (HaCaT) and to understand its potential mechanisms on ageing. Methods　HaCaT cells were cultured in vitro and divided into control group, H2O2 model group, and SchC treatment group. Cell viability was evaluated by CCK-8 assay. The levels of superoxide dismutase (SOD), malondialdehyde (MDA) and glutathione (GSH) were determined by WST-1 assay. The level of reactive oxygen species (ROS) was determined by chemical fluorescence assay. The mRNA expression of matrix metalloproteinase, cyclooxygenase-2 (COX-2) and apoptosis-related genes were detected by qRT-PCR. The expression of COX-2, matrix metalloproteinase, apoptosis-related protein, ageing-related protein, transcription factor NF-E2 related factor 2 (Nrf-2), heme oxygenase (HO-1), cytoplasmic transcription factor-κB (NF-κB) and p-NF-κB was detected by Western blotting. Results　 SchC at the concentration of less than 100 μmol/L had no significant effects on the proliferation of HaCaT cells; while cell viability decreased to (57.0±3.0)% (P<0.001) after treated with 800 μmol/L H2O2. Compared with the H2O2 model group, the cell viability, SOD, and GSH levels were significantly increased; MDA and ROS levels were significantly decreased (P<0.05 or P<0.01). Western blotting results showed that the expression of P16, P21 and p-NF-κB were significantly down-regulated, and the expression of Nrf-2 and HO-1 were up-regulated by SchC (P<0.05 or P<0.01). qRT-PCR and Western blotting results showed that SchC could upregulate the expression of caspase-3 and caspase-9, and down-regulate the expression of Bcl-2, matrix metalloproteinase (MMP-1, MMP-9) and inflammatory factor COX-2 by regulating NF-κB pathway (P<0.05 or P<0.01). Conclusion　SchC had a protective effect on the aging model of oxidative stress damage of H2O2-induced HaCaT cells, which could lay a foundation for the development of anti-ageing products of Schisandra C.