β-chemokine function in experimental lung ischemia-reperfusion injury

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β-chemokine function in experimental lung ischemia-reperfusion injury Baiya Krishnadasan, MD, Alexander S. Farivar, MD, Babu V. Naidu, FRCS, Steven M. Woolley, MRCS, Karen Byrne, BS, Charles H. Fraga, MS, Michael S. Mulligan, MD The Annals of Thoracic Surgery Volume 77, Issue 3, Pages 1056-1062 (March 2004) DOI: 10.1016/S0003-4975(03)01600-X

Fig 1 Western blot analysis of left lung homogenates. The far left lane is the molecular marker, representing weight (MW) in kilodaltons (Kda). The next 3 lanes from the left represent unmanipulated controls and lungs subjected to ischemia and up to 2 hours of reperfusion in which there was no detectable expression of MIP-1α, MCP-1, or RANTES protein. There was increased protein expression of MIP-1α at 3 and 4 hours of reperfusion. MCP-1 and RANTES protein were weakly detectable at 4 hours of reperfusion. (MCP = monocyte chemoattractant protein; MIP = macrophage inflammatory protein; RANTES = regulated upon activation, normal T cells expressed and secreted.) The Annals of Thoracic Surgery 2004 77, 1056-1062DOI: (10.1016/S0003-4975(03)01600-X)

Fig 2 Northern blot analysis detected MIP-1α messenger RNA (mRNA) expression at 30 minutes of reperfusion, and this continued throughout the reperfusion period. There was a relative decrease in mRNA expression at 3 and 4 hours of reperfusion compared with the earlier time points. There was no mRNA expression in unmanipulated controls. (C = control; GAPDH = glyceraldehyde-3-phosphate dehydrogenase; MIP = macrophage inflammatory protein.) The Annals of Thoracic Surgery 2004 77, 1056-1062DOI: (10.1016/S0003-4975(03)01600-X)

Fig 3 Ribonuclease protection assay. Three experimental groups were studied and include both the right and left lung for each. The left (L) and right (R) lungs of unmanipulated negative controls are shown in the far left lanes. The middle 2 lanes represent the lungs of animals that underwent ischemia and 4 hours of reperfusion, and the far right lanes represent the lungs of animals that underwent lung ischemia-reperfusion injury and that were treated with antibodies to macrophage inflammatory protein-1α. Relative to the positive control lungs in the middle 2 lanes, there was a dramatic decrease in both IL-2 and TNF-α messenger RNA (mRNA) expression in the injured left lungs of animals treated with antibodies to macrophage inflammatory protein-1α, whereas the mRNA of the anti-inflammatory peptide IL-4 was enhanced. There was relatively minimal transcriptional activation in the right lungs of all groups. (IL = interleukin; TNF = tumor necrosis factor.) The Annals of Thoracic Surgery 2004 77, 1056-1062DOI: (10.1016/S0003-4975(03)01600-X)