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Figure 4 | Critical Care

Figure 4

From: Endothelial dysfunction is a potential contributor to multiple organ failure and mortality in aged mice subjected to septic shock: preclinical studies in a murine model of cecal ligation and puncture

Figure 4

Assessment of oxidative damage and endothelial dysfunction in aortas isolated from septic mice. (Panel A) Following CLP, young, aged and eNOS-/- mice were euthanized by opening the chest cavity under deep anesthesia and the thoracic aortas were isolated. Twenty μg of protein extracts were subjected to SDS-PAGE and derivatized protein carbonyl groups were immunodetected by OxyBlot (Merck Millipore). Immunoreactivity was visualized by chemiluminescent detection. Blots were scanned and a densitometric analysis was performed (Panel B) (*P <0.05 vs. young mice; n = 3). In a separate set of experiments, aortic rings (approximately 2 mm in length) were harvested from sham-operated and CLP-injured mice, placed in 5 ml organ baths filled with oxygenated (95% O2 to 5% CO2) Krebs-Henseleit solution at 37°C and mounted onto isometric force transducers. Concentration response curves to acetylcholine (Ach) (Panel C and D) and sodium nitroprusside (SNP) (Panel D and E) were performed on a stable phenylephrine-induced tone. Please notice the inability of the aortic rings from eNOS-/- mice to respond to Ach stimulation. Aged mice showed a basal level of endothelial dysfunction (Panel C) (*P <0.05 vs. young). CLP induced a shift to the right of the concentration response curve to Ach, which was more pronounced in the aged mice compared to the young (Panel D) (mean LogEC50 from -7.797 to -7.488 in young mice; mean LogEC50 from -7.261 to -6.762 in aged mice). Increased sensitivity to the nitric oxide (NO) donor SNP was also seen in aged and eNOS-deficient vessels (Panel E and F). CLP, cecal ligation and puncture; eNOS, endothelial isoform of nitric oxide synthase.

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