Neuroprotective Effect of Salvianolic Acid A against Diabetic Peripheral Neuropathy through Modulation of Nrf2

Figure 8

Salvianolic acid A exerted the protective effect on high glucose-injured RSC96 cells through modulation of Nrf2. (a) Molecular interaction of SalA with the active sites of the Keap1 Kelch domain using the program LibDock of Discovery Studio software. (b) RSC96 cells were transfected with pGL4-ARE along with a promoterless Renilla luciferase construct using the jetPRIME reagent and then treated with 150 mM glucose and different concentrations of SalA. The promoter activity of luciferase was detected by a reporter assay kit. All data are presented as , . , , and , compared with the HG group. (c) qPCR analysis of the Nrf2 gene expression in RSC96 cells treated with 150 mM glucose and different concentrations of SalA for 24 h. All qPCR data are normalized to TBP and presented as , . , , , compared with the HG group. (d) Representative blot image and quantitative analysis ( western blot) of total Keap1, total Nrf2, total β-actin (loading control), nuclear Nrf2, histone H3 (loading control), cytoplasm Nrf2, and cytoplasm β-actin (loading control) protein in RSC96 cells treated with 150 mM glucose and different concentrations of SalA for 24 h from independent experiments. RSC96 cells were treated with 150 mM glucose and different concentrations of SalA, followed by staining using Nrf2 antibody. The cells were scanned by a high content screening assay (scale  μm) (e), and average relative fluorescence was quantified by the intensity of fluorescence from individual cells (f). (g) RSC96 cells were transfected with control SiRNA or Nrf2 SiRNA using a jetPRIME reagent. 48 h after transfection, the cells were incubated with 150 mM glucose and different concentrations of SalA. After 24 h treatment, the cell viability was detected with a CCK-8 assay. All data are presented as , . , , and , compared with the HG group.

(a)

(b)

(c)

(d)

(e)

(f)

(g)