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. 2016 Jun 9;65(9):2553–2568. doi: 10.2337/db16-0387

Figure 8.

Figure 8

MCAT does not improve recovery from hindlimb ischemia in chow-fed mice. A: Representative LDPI images pre-HLI and 7 days post-HLI (d7). B: Quantification of LDPI flux demonstrates that paw perfusion recovery was not different between WT mice (n = 4) and MCAT mice (n = 4) on chow diet. C: High-resolution respirometry experiments performed on isolated skeletal muscle mitochondria show that the respiratory capacity was not different between WT and MCAT mice (n = 4/group) and ischemia induced a small, nonsignificant decrease in respiration. D: Mitochondrial H2O2 emission under state 4 conditions with 25 μmol palmitoyl carnitine, 10 mmol glutamate, 2 mmol malate, 10 mmol succinate, and 10 mmol glycerol-3-phosphate was substantially lower in both control and ischemic limbs of MCAT mice, consistent with increased scavenging by catalase overexpression (n = 4/group). E: Free radical leak (%), calculated by normalizing data from D by state 4 JO2 measured in parallel experiments under identical substrate conditions, was also lower in the control and ischemic limbs of MCAT mice compared with those of WT mice (n = 4/group), but no ischemia effects were detected in either genotype. F: Proportions of necrosis scores at HLI day 7 for WT and MCAT mice on chow diet (n = 4/group). G: Representative H&E stains of the control and ischemic limbs of WT and MCAT mice fed a chow diet. Images were acquired at 20× magnification. Data are mean ± SEM and were compared with two-way ANOVA followed by Tukey posttest or two-tailed Student t test. Nonparametric data (necrosis scores) were compared with Mann-Whitney test. aP < 0.05 for genotype effect, bP < 0.05 for ischemia effect. AmA, antimycin A; Complex I(state 3), glutamate + malate + adenosine diphosphate; Complex I(state 4), glutamate + malate; Complex I+II(state 3), glutamate + malate + adenosine diphosphate + succinate; Complex II(state 3), glutamate + malate + adenosine diphosphate + succinate + rotenone; Complex IV, ascorbate + N,N,N',N'-tetramethyl-p-phenylenediamine; Cyt c, cytochrome c; G, glutamate; G3P, glycerol-3-phosphate; M, malate; PC, palmitoyl carnitine; Succ, succinate; Uncoupled, carbonyl cyanide-4-(trifluoromethoxy)phenylhydrazone.