CMS Activated SIRT1 Expression in a STZ Cell Model
Next, the relationship between CMS and SIRT1 was investigated. The
results of RT-qPCR and western blot analysis demonstrated that
expression of SIRT1 in ARPE-19 cells transfected with sh-SIRT1-1,
sh-SIRT1-2, or sh-SIRT1-3 all declined dramatically with sh-SIRT1-2
exhibiting the highest effect (Figure 3A - B), therefore
sh-SIRT1-2 was selected for the
following study. Immunofluorescence staining revealed that stimulation
with HG-FFA notably decreased activity and nuclear accumulation of
SIRT1; also, the activity and nuclear accumulation of SIRT1 were
significantly higher in cells treated with CMS than in cells stimulated
with HG-FFA, the effect of CMS on SIRT1 was dose-dependent, and the
increase in cells treated with 2-CMS was the highest (Figure 3C - D).
According to the result of RT-qPCR, compared with the controls, the
expression of SIRT1 was decreased in ARPE-19 cells stimulated with
HG-FFA, while the expression of SIRT1 was notably higher in cells
stimulated with CMS than in cells stimulated with HG-FFA, the effect of
CMS on SIRT1 was dose-dependent, and the increase in cells treated with
2-CMS was the highest (Figure 3E). Collectively, CMS could activate the
expression of SIRT1 in a STZ cell model, and sh-SIRT1-2 and 2-CMS were
selected for the following study.