Activation of Nrf2/Keap1 signaling and autophagy induction against oxidative stress in heart in iron deficiency

Figures & data

Table 1. Body weight; food intake; hemoglobin concentration; heart, liver, and kidney weights in mouse fed a control diet or an iron-deficient diet.

	Control	Iron-deficient
Initial body weight (g)	18.97 ± 0.76	18.99 ± 0.78
Initial body weight (g)	27.76 ± 0.85	26.12 ± 0.55
Food intake (g/28 days)	131.83 ± 4.2	118.08 ± 2.8
Hemoglobin (g/dl)	14.51 ± 4.2	8.55 ± 2.79^a
Heart weight (g/100 g body weight)	0.44 ± 0.04	0.51 ± 0.01^a
Kidney weight (g/100 g body weight)	1.55 ± 0.05	1.49 ± 0.1
Liver weight (g/100 g body weight)	5.11 ± 0.51	5.17 ± 0.52

Note:

Data are presented as the means ± SEM for each group of eight mice.

^a Significant differences from the control group at p < 0.05.

Fig. 1. Effects of iron-deficiency on oxidation of proteins and autophagy.

Notes: (A) Mouse heart tissues and per mice were homogenized with fourfold of homogenization buffer, after which Western blot analyses were performed to detect the carbonyl proteins using anti-DNP antibody and an autophagic marker, LC3-II antibody. (B) Western blots of 100 μM DFO-treated and DFO-untreated NIH 3T3 cells for 24 h lysates probed for DNP and LC3-II. β-Actin was used as internal control.

Fig. 2. Effect of iron deficient on Nrf2 and its target genes in control and iron deficient mice in heart.

Notes: (A) Heart tissues of CTL and ID group were prepared nuclear fraction and subjected to Western blot against anti-Nrf2 antibody. Histone was used as nuclear marker. (B), (C), and (D), After total RNA of heart tissue was extracted using TRIzol reagent, cDNA was synthesized using 1 μg of total RNA and an RNA PCR AMV kit (Takara Bio, Shiga, Japan). Then, the expression levels of HO-1 (B), CuZnSOD (C), and MnSOD (D) mRNA were quantified by qRT-PCR, with β-actin serving as the internal standard. Each value represents the mean ± SEM.