Oxidative stress induces increases in the gene expression and protein levels of SIRT2 in primary murine astrocyte cultures and PC12 cells

Abstract

Oxidative stress is a major pathological factor in multiple major neurological diseases. Certain Sirtuin family proteins, such as sirtuin 1 (SIRT1), also play important roles in aging and cell survival. In this study we investigated the effect of oxidative stress on the gene expression and protein levels of sirtuin 2 (SIRT2) in primary murine astrocyte cultures and PC12 cells. Our Western blot studies, using the samples collected at 24 hrs after H2O2 treatment, showed that treatment of primary astrocyte cultures with 0.1, 0.3 and 0.5 mM H2O2 increased SIRT2 levels by 100% ‐‐‐ 300% in astrocytes; and treatment of PC12 cells with 0.1, 0.3 and 0.5 mM H2O2 increased SIRT2 levels by 30% ‐‐‐ 80% in the cells. Our real‐time PCR study, using the samples collected at 12 hrs after H2O2 treatment, has also suggested that 0.1 and 0.5 mM H2O2 increased the SIRT2 mRNA levels by approximately 100% in astrocytes. Our immunocytochemistry study has further provided preliminary results suggesting that H2O2 can induce an increase in SIRT2 levels in PC12 cells. Collectively, our study has suggested that oxidative stress can significantly increase the gene expression and protein levels in primary astrocyte cultures and PC12 cells, which may be involved in oxidative stress‐induced cell injury (supported by a National Key Basic Research ‘973 Program Grant’ #2010CB834306, a Pujiang Scholar Program Award 09PJ1405900, and a Key Research Grant of Shanghai Municipal Educational Committee #09ZZ21).