Research Paper Volume 11, Issue 18 pp 7746—7779

Inadequate control of thyroid hormones sensitizes to hepatocarcinogenesis and unhealthy aging

Figure 6. Mild hypothyroidism produces mitochondrial dysfunction and accumulation of oxidative damage in Pax8 +/- mice. (A) mRNA expression of genes involved in mitochondrial biogenesis and mitochondrial function in liver. Pgc1-α: n = 6 per group. Ndufab5 = n = 6 per group. Uqcrc1: n = 6 per group. Cox5b: n = 6 per group. Atp5a1: n = 5 Wt, n = 6 Pax8 +/-. Ucp2: n = 5 per group. (B) Western blots showing expression levels of proteins involved in mitochondrial biogenesis/function as well as Ampk and its phosphorylated isoform in liver lysates. n = 6 per group. (C) Densitometric analysis of western blots shown in panel B. n = 6 per group. (D) Relative mitochondrial DNA content in liver isolations. n = 6 per group. (E) Citrate synthase activity in liver extracts. n = 6 per group. (F) Complex I+III activity in liver extracts. n = 6 per group. (G) Complex I+III activity corrected by citrate synthase activity in liver extracts. n = 6 per group. (H) Complex II+III activity in liver extracts. n = 6 per group. (I) Complex II+III activity corrected by citrate synthase activity in liver extracts. n = 6 per group. (J) Superoxide generation in mitochondrial complexes in liver extracts. n = 6 per group. (K) Superoxide generation in mitochondrial complexes corrected by citrate synthase activity in liver extracts. n = 6 per group. (L) Oxygen consumption rate in primary hepatocytes. n = 3 per group. (M) Extracellular acidification rate in primary hepatocytes. n = 3 per group. (N) MTT test to determine the metabolic activity (NADH-oxidase) of primary hepatocytes. n = 4 per group. (O) Antioxidant enzymatic activities in liver lysates. Sod mit: n = 6 per group. Sod cyt: n = 5 per group. Cat: n = 5 Wt, n = 6 Pax8 +/-. Gst: n = 5 per group. Gpx: n = 5 per group. Grd: n = 6 Wt, n = 5 Pax8 +/-. Nqo1: n = 5 Wt, n = 6 Pax8 +/-. Sod mit: Sod mitochondrial. Sod cyt: Sod cytosolic. (P) Western blots showing expression levels of antioxidant, stress response and autophagy induction proteins in liver lysates. LE = Low exposure. HE = High exposure. n = 6 per group. (Q) Densitometric analysis of western blots shown in panel P. ® Western blot showing Lys-4-HNE staining in liver extracts. n = 6 per group. (S) Densitometric analysis of the western blot shown in panel R. Mice were 9 month-old at the time of killing. Data are represented as the mean ± SEM. * p-value < 0.05. T-test two tailed.