Fig. 5

TNF-α led to neuronal mitochondrial dysfunction via downregulating the AMPK-Sirt3 pathway. A Representative immunofluorescence images and quantitative analysis showed colocalization of Sirt3 and tyrosine hydroxylase (TH)-positive neurons in the RVLM of control and SIH groups. Scale bar = 500 or 20 μm. B Representative immunoblot bands and quantitative analysis showed the expression of p-AMPK and Sirt3 in the RVLM of control and SIH rats. C Representative immunoblot bands and quantitative analysis of p-AMPK and Sirt3 expression in N2a cells treated with or without TNF-α. D Representative images and quantitative analysis showed mitochondrial membrane potential (MMP) in control, TNF-α, and TNF-α + A769662 groups. Scale bar = 50 μm. E Representative immunoblot bands and quantitative analysis showed the expression of mitochondrial respiratory chain complexes (complexes II-SDHB, III-UQCRC2, and V-ATP5A) in N2a cells with different treatments. F Representative fluorescence images of reactive oxygen species (ROS) detection and statistical data of ROS production in N2a cells with or without TNF-α treatment. Scale bar = 50 μm. G The levels of superoxide dismutase (SOD) and catalase (CAT) were quantified by using commercial kits in each group. Data were shown as mean ± SEM. Statistical significance was determined using two-tailed unpaired Student’s t-test (A–C) and one-way ANOVA followed by post hoc Bonferroni test (D–G). n = 12 slices from 6 rats, two slices per rat (A). n = 3 rats per group (B). n = 3 of independent cell culture preparations (C, E). n = 6 of independent cell culture preparations (G). n = 12 slices from 6 of independent cell culture preparations, two slices per independent cell culture preparation (D, F). *p < 0.05 vs. SIH group. ^#p < 0.05, ^##p < 0.01  vs. TNF-α group