CMS Inhibited Oxidative Stress and Inflammation of Retinal Cells through Activating SIRT1
In order to evaluate whether CMS regulated oxidative stress and inflammation in retinal cells with DR through SIRT1, we silenced SIRT1 expression in ARPE-19 cells. ELISA was performed to detect the expression of oxidative stress-related factors, which observed that CMS treatment obviously reduced the expression of ROS and MDA, but increased SOD expression in ARPE-19 cells stimulated with HG-FFA, which could be blocked by sh-SIRT1 (Table 3). In addition, the results of RT-qPCR and western blot analysis demonstrated that iNOS expression in ARPE-19 cells stimulated with HG-FFA was declined by CMS treatment, which could be reversed by sh-SIRT1 (Figure 4A - B). Nitrite test revealed that CMS treatment notably decreased the expression of NO in HG-FFA stimulated ARPE-19 cells, which could be rescued by sh-SIRT1 (Figure 4C). According to the result of ELISA, in ARPE-19 cells stimulated with HG-FFA, notably decreased expression of IL-6, TNF-α, and CRP was observed after CMS treatment, which could be reversed by sh-SIRT1 (Figure 4D - F). Therefore, CMS was beneficial to alleviate the oxidative stress and inflammation in retinal cells of DR through activating SIRT1.