Mitochondrial dysfunction in patients with celiac disease: A multidimensional study

Figure 1 Transmission electron microscopy images of duodenal intestinal epithelial cells from celiac disease patients and healthy controls (n = 3 per group). A-F: Healthy control group (A-C: × 12000; scale bars = 2 μm. D-F: × 20000; scale bars = 1 μm); G-L: Celiac disease group (G-I: × 12000; scale bars = 2 μm. J-L: × 20000; scale bars = 1 μm). Blue arrows indicate swollen mitochondrial matrix; pink arrows indicate cristae disintegration, disruption, or loss; yellow arrows indicate pleomorphic mitochondria. HC: Healthy control; CeD: Celiac disease.

Figure 2 Changes in mitochondrial biochemical indicators, apoptosis, and mitochondrial DNA copy number. A: Adenosine triphosphate content; B: L-lactate levels; C: 4-hydroxynonenal content; D: Malondialdehyde content; E: Superoxide dismutase activity; F: Glutathione peroxidase activity; G: Mitochondrial DNA copy number; H: Caspase 9 activity. Data are shown as mean ± SD or median (IQR) as appropriate. P values were determined by paired t-test or Wilcoxon signed-rank test. Group sizes: n = 15 per group. ATP: Adenosine triphosphate; 4-HNE: 4-hydroxynonenal; MDA: Malondialdehyde; SOD: Superoxide dismutase; GSH-PX: Glutathione peroxidase; mtDNA: Mitochondrial DNA; HC: Healthy control; CeD: Celiac disease.

Figure 3 Expression of mitochondrial fusion and fission proteins (n = 3 per group for western blotting). A: Western blot analysis showed that the fusion proteins OPA1, MFN1, and MFN2 were expressed in the intestinal mucosa of the celiac disease group; B-D: Quantitative analysis of the fusion proteins OPA1 (B), MFN1 (C), and MFN2 (D); E: Western blot analysis showed that the fission-promoting proteins DRP1, p-DRP1 (the activated form), and FIS1 were expressed in the intestinal mucosa of the celiac disease group; F-H: Quantitative analysis of the fission-promoting proteins p-DRP1 (F), DRP1 (G), and FIS1 (H). Bar charts represent relative protein expression normalized to β-actin (n = 3 per group). Data are presented as mean ± SD. β-actin was used as a loading control. HC: Healthy control; CeD: Celiac disease.

Figure 4 Correlation between mitochondrial damage indicators and their association with clinical features. Scatter plots show the following significant correlations (after Benjamini-Hochberg false discovery rate correction, q < 0.10). A: Adenosine triphosphate levels with superoxide dismutase activity; B: L-lactate levels with caspase 9 activity; C: 4-hydroxynonenal concentration with serum 25-hydroxyvitamin D levels; D: L-lactate levels with erythropoietin; E: L-lactate levels with serum ferritin; F: Caspase 9 activity with erythropoietin. The Spearman correlation coefficient (r) and P value are indicated in each plot, with the solid line representing the linear regression fit (n = 15 per group). SOD: Superoxide dismutase; ATP: Adenosine triphosphate; 4-HNE: 4-hydroxynonenal; 25(OH)D: 25-hydroxyvitamin D.