Figure 6
Differences between MitoQ and rampril treated diabetic kidney cortices. White bars – Non-diabetic db/m mice (C); Black bars/dots diabetic db/db mice (D); Blue bars/triangles diabetic db/db mice + 0.6 mg/kg/day MitoQ intragastrically (DMitoQ); Red bars/squares diabetic db/db mice +3 mg/kg/day ramipril intragastrically (DRam); Grey bars/dots - diabetic db/db mice + combination of MitoQ and ramipril (DCoAd). n = 6 mice/group. (A) Scores plot of the bioinformatic (OPLS-DA) model including all renal metabolites, showing clear separation among diabetic mouse groups where the predictive component 1 axis is for untreated diabetic (D) mice vs ramipril treated mice are shown (x axis, PC1), plotted against the predictive component 2 axis for untreated diabetic mice vs MitoQ treated mice (y axis, PC2). (B) Table of the most influential metabolomic variables which define the differences between treated and untreated diabetic mouse groups. The variable importance on prediction (VIP) parameter ranks the metabolites in order of significance. Renal cortical concentrations of the metabolites which differ between MitoQ and ramipril treated mouse groups, (C) dihydroxyacetone phosphate (DHAP) and glucose 1-phosphate (G1P), (D) Pearson’s correlations between renal function defined as GFR and G1P/DHAP in all mice. (E) succinate:fumarate ratio, F-h SeaHorse XF24 Flux Analyses of oxygen consumption rate (OCR) during a maximal electron flow test in isolated uncoupled renal cortical mitochondria in the presence of FCCP and the complex I substrates, pyruvate and malate. n = 3 in the db/m and n = 5 mice/group in the db/db with and without treatment. Line traces of OCR are shown for (F) control vs diabetic untreated mice and (G) diabetic MitoQ vs ramipril treated mouse groups - the curve for the diabetic untreated group sits below the ramipril group. (H) Mitochondrial OCR responses at baseline, following addition of rotenone (CI inhibitor), succinate (CII substrate), antimycin A (CIII) inhibitor and ascorbic acid + TMPD − N,N,N′,N′-tetramethyl-p-phenylenediamine (Asc/TMPD; CIV substrates) of the mitochondrial respiratory chain. For electron flow, all data are expressed as median ± interquartile range. *P < 0.05 vs C; † P < 0.05 vs D; ‡ P < 0.05 vs DMitoQ all by 1 W ANOVA/Tukey’s Post-hoc.
