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Simvastatin ameliorates injury in an experimental model of lung ischemia-reperfusion  Babu V Naidu, FRCS, Steven M Woolley, MRCS, Alexander S Farivar,

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Presentation on theme: "Simvastatin ameliorates injury in an experimental model of lung ischemia-reperfusion  Babu V Naidu, FRCS, Steven M Woolley, MRCS, Alexander S Farivar,"— Presentation transcript:

1 Simvastatin ameliorates injury in an experimental model of lung ischemia-reperfusion 
Babu V Naidu, FRCS, Steven M Woolley, MRCS, Alexander S Farivar, MD, Robert Thomas, BA, Charles Fraga, MSc, Michael S Mulligan, MD, FACS  The Journal of Thoracic and Cardiovascular Surgery  Volume 126, Issue 2, Pages (August 2003) DOI: /S (03)

2 Figure 1 Vascular permeability. Treatment with simvastatin before ischemia-reperfusion showed a significant reduction in vascular permeability at 15 minutes (P = .01, n = 3) and 4 hours (P < .001, n = 3) of reperfusion. The Journal of Thoracic and Cardiovascular Surgery  , DOI: ( /S (03) )

3 Figure 2 Neutrophil recruitment. A, Tissue neutrophil accumulation as measured by myeloperoxidase (MPO) content was statistically increased at 4 hours of reperfusion when compared with negative controls (P < .001). These values were markedly decreased in statin-treated animals (P < .002, n = 4). B, The bronchoalveolar lavage (BAL) cell counts increased 26-fold in animals undergoing ischemia-reperfusion when compared with negative controls (P < .001). A significant decrease in leukocyte accumulation was noted in BAL from animals receiving statin (P < .001, n = 4). The Journal of Thoracic and Cardiovascular Surgery  , DOI: ( /S (03) )

4 Figure 3 Reversal of statin protection with NG-nitro-l-arginine methyl ester (l-NAME). There was no significant difference in lung vascular permeability between untreated animals and animals treated with l-NAME after 4 hours of reperfusion. The decrease in permeability associated with statin treatment was reversed with concurrent l-NAME administration (P < .03). The Journal of Thoracic and Cardiovascular Surgery  , DOI: ( /S (03) )

5 Figure 4 Enzyme-linked immunosorbent assay for tumor necrosis factor (TNF)-α content of lavage fluid. There was a marked increase in TNF-α content in lavage fluid from left lungs at 4 hours of reperfusion. Statin-treated animals demonstrated a significant reduction of TNF-α when compared with positive controls (P < .013; n = 4). The Journal of Thoracic and Cardiovascular Surgery  , DOI: ( /S (03) )

6 Figure 5 Electrophoretic mobility shift assay for nuclear factor (NF)-κB and activator protein (AP)-1. The first 2 columns represent unmanipulated negative controls. The middle 2 columns are positive controls subjected to ischemia and 15 minutes of reperfusion. The final 2 columns reveal a marked reduction in nuclear translocation of NF-κB and AP-1 in animals treated with simvastatin (n = 4). The Journal of Thoracic and Cardiovascular Surgery  , DOI: ( /S (03) )

7 Figure 6 Western blot for endothelial nitric oxide synthase (eNOS) expression. eNOS protein, with ischemia and 15 minutes of reperfusion, is increased above baseline in both treated and nontreated groups. Conversely, eNOS expression persists only in the statin-treated group at 4 hours of reperfusion. The Journal of Thoracic and Cardiovascular Surgery  , DOI: ( /S (03) )

8 Figure 7 Nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase stain. Statin treatment reduces NADPH-dependent oxidase activity in left lungs at 15 minutes of reperfusion when compared with nontreated controls. Frozen sections stained are as follows. A, Normal left lung (×200). B, Nontreated left lungs subjected to ischemia and 15 minutes of reperfusion (×200). C, Simvastatin pretreatment followed by ischemia and 15 minutes of reperfusion (×200). The Journal of Thoracic and Cardiovascular Surgery  , DOI: ( /S (03) )

9 Figure 8 Characterization of NADPH oxidase activity. Nondenaturing electrophoresis of whole lung protein lysates were performed on 6% polyacrylamide gels. Gels were stained for enzyme activity using a modified diaphorase. A, Prestained standard. Myosin (200 kDa) and β-galactosidase (133 kDa). B, Unmanipulated lungs. C, Lungs from statin-treated unmanipulated animals. D, Animals subjected to ischemia and 15 minutes of reperfusion. Contains a single band (calculated molecular weight 253 kDa) that correlates to intact NADPH oxidase (molecular weight 267 kDa). E, Statin-treated animals subjected to ischemia and 15 minutes of reperfusion. Statin pretreatment resulted in a dramatic reduction in NADPH oxidase activity. The Journal of Thoracic and Cardiovascular Surgery  , DOI: ( /S (03) )

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