Nitric oxide, enhanced by macrophage-colony stimulating factor, mediates renal damage in reflux nephropathy Udo Rolle, Hideki Shima, Prem Puri Kidney.

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Nitric oxide, enhanced by macrophage-colony stimulating factor, mediates renal damage in reflux nephropathy Udo Rolle, Hideki Shima, Prem Puri Kidney International Volume 62, Issue 2, Pages 507-513 (August 2002) DOI: 10.1046/j.1523-1755.2002.00485.x Copyright © 2002 International Society of Nephrology Terms and Conditions

Figure 1 NADPH-diaphorase (NADPH-d) histochemistry. (A) Normal kidney shows weak NADPH-d activity in the epithelium of proximal and distal tubules. (B) Reflux nephropathy (RN) shows strong NADPH-d activity in the epithelium of distal tubules. Kidney International 2002 62, 507-513DOI: (10.1046/j.1523-1755.2002.00485.x) Copyright © 2002 International Society of Nephrology Terms and Conditions

Figure 2 NADPH-diaphorase (NADPH-d) histochemistry/macrophage-colony stimulating factor (M-CSF) immunohistochemistry double staining. (A) Normal kidney where NADPH-d activity/M-CSF immunoreactivity is absent in the epithelium of proximal and distal tubules. (B) RN kidney showing weak M-CSF immunoreactivity in the epithelium of proximal and strong M-CSF immunoreactivity colocalized with moderate NADPH-diaphorase activity in distal tubules. Kidney International 2002 62, 507-513DOI: (10.1046/j.1523-1755.2002.00485.x) Copyright © 2002 International Society of Nephrology Terms and Conditions

Figure 3 Macrophage-colony stimulating factor (M-CSF)/inducible nitric oxide synthase (iNOS) double immunohistochemistry using confocal microscopy. Positive labeling for M-CSF appears in red color, positive labeling for iNOS appears in green color, and double labeling results in a yellow color. (A) Normal kidney where there is no expression in the epithelium of any tubules and Bowman's capsule. (B) RN kidney showing strong expression of M-CSF/iNOS double immunohistochemistry in distal tubules. Kidney International 2002 62, 507-513DOI: (10.1046/j.1523-1755.2002.00485.x) Copyright © 2002 International Society of Nephrology Terms and Conditions

Figure 4 Inducible nitric oxide synthase (iNOS) mRNA expression by reverse transcription-polymerase chain reaction (RT-PCR). Lanes 1 to 3 are expressed from patients with RN and lanes 4 to 7 are from normal controls. The PCR condition successfully yielded the amplified fragments of expected size for iNOS and G3PDH in each samples. Intensity of the bands corresponding to G3PDH mRNA was approximately similar among the samples. mRNA levels of iNOS are increased in refluxing kidneys (lanes 1-3) compared to normal kidneys (lanes 4-7). Kidney International 2002 62, 507-513DOI: (10.1046/j.1523-1755.2002.00485.x) Copyright © 2002 International Society of Nephrology Terms and Conditions

Figure 5 Apoptosis. (A) Normal kidney shows few apoptotic cells in the epithelium of proximal and distal tubules, whereas the (B) RN kidney shows an increased number of apoptotic cells in the epithelium of the distal tubules and (C) a markedly increased number of apoptotic cells in the epithelium of collecting ducts. Kidney International 2002 62, 507-513DOI: (10.1046/j.1523-1755.2002.00485.x) Copyright © 2002 International Society of Nephrology Terms and Conditions