Mitochondrial Alterations and Enhanced Human Leukocyte/Endothelial Cell Interactions in Type 1 Diabetes

Figures

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  Figure 1. Evaluation of oxidative stress (total and mitochondrial ROS production) and mitochondrial membrane potential in type 1 diabetic and control subjects. (A) Levels of total ROS production measured as DCFH-DA fluorescence. Representative images of leukocytes stained with DCFH-DA (green) and Hoechst 33,342 (blue) and visualized by fluorescence microscopy. (B) Levels of mitochondrial ROS production measured as MitoSOX red fluorescence. Representative images of leukocytes stained with MitoSOX (red) and Hoechst 33,342 (blue) by fluorescence microscopy. (C) Mitochondrial membrane potential measured as TMRM fluorescence. Representative images of leukocytes stained with TMRM (red) and Hoechst 33,342 (blue) and visualized by fluorescence microscopy. Fluorescence was measured as arbitrary units of fluorescence and expressed as % of control fluorescence. Data are expressed as mean ± SEM. * p < 0.05 control vs. type 1 diabetic subjects.

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  Figure 2. Evaluation of PMNs interactions in type 1 diabetic and control subjects, and levels of adhesion molecules in serum. (A) PMNs rolling velocity (μmsecond−1); (B) PMN rolling flux (cells/min); (C) PMNs adhesion (cells/mm2); (D) Levels of P-selectin (ng/mL); (E) Levels of VCAM-1 (ng/mL); (F) Levels of ICAM-1 (ng/mL). Data are expressed as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 control vs. type 1 diabetic subjects. Abbreviations: ICAM-1: intercellular adhesion molecule-1; PMNs, polymorphonuclear leukocytes; P-selectin: platelet selectin; VCAM-1: vascular cell adhesion molecule-1.

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  Figure 3. Evaluation of serum levels of cytokines and mediators of inflammation in type 1 diabetic and control subjects. (A) Serum levels of TNFα (pg/mL); (B) Serum levels of IL-6 (pg/mL); (C) Serum levels of MPO (pmol/L). Data are expressed as mean ± SEM. * p < 0.05, ** p < 0.01 control vs. type 1 diabetic subjects. Abbreviations: IL-6, interleukin 6; MPO, myeloperoxidase; TNFα: tumor necrosis factor alpha.

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  Figure 4. Evaluation of mRNA expression of SOD (A) and CAT (B) in type 1 diabetic and control subjects. Data are expressed as mean ± SEM. * p < 0.05, control vs. type 1 diabetic subjects. * p < 0.05 control vs. type 1 diabetic subjects.