Idh2 Deficiency Exacerbates Acrolein-Induced Lung Injury through Mitochondrial Redox Environment Deterioration

Figure 4

Acrolein-induced lung damage in idh2^−/− mice. Mice were exposed to filtered air (control) or acrolein (10 ppm, 12 h). (a) Immunoblot analysis of IDH2 protein expression using an anti-IDH2 antibody. β-Actin was used as an internal control. (b) H&E-stained sections of the lung tissues after acrolein exposure. (c) Emphysema in acrolein-treated lung tissues assessed by mean alveolar airspace area (μm²). (d) Lung injury scores were evaluated after acrolein exposure. (e) TUNEL staining of the lung tissues from acrolein-treated idh2^−/− and WT (idh2^+/+) mice. (f) Immunoblots comparing the levels of apoptotic marker proteins in the lung tissue extracts from acrolein-treated idh2^−/− and WT mice. β-Actin was used as an internal control. (g) Intracellular H₂O₂ was measured using xylenol orange. (h) Immunoblot analysis of Prx-SO₃ levels in the lung tissue extracts from WT and idh2^−/− mice. (i) The levels of MDA accumulated in the lung tissue extracts were determined using TBARS assay. (j) The levels of acrolein-adducted proteins in the lung tissue extracts were measured with anti-acrolein antibody. (k) The levels of oxidized protein adducts, (l) glutathionylated protein adducts, and (m) nitrosylated protein adducts in the lung tissue extracts were measured with anti-DNP, anti-GSH, and anti-nitrotyrosine antibodies, respectively. In (c), (d), (g), and (i), data are shown as the mean ± SD ( mice in each group). versus acrolein-treated WT mice.

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