Yonsei Med J.  2007 Jun;48(3):517-525. 10.3349/ymj.2007.48.3.517.

Activation of Intrarenal Complement System in Mouse Model for Chronic Cyclosporine Nephrotoxicity

• Kim YO
• Lim SW
• Li C
• Kang HJ
• Ahn KO
• Yang HJ
• Ghee JY
• Kim SH
• Kim JY
• Choi BS
• Kim J
• Yang CW

• KMID: 724109
• DOI: http://doi.org/10.3349/ymj.2007.48.3.517

Abstract

PURPOSE
Local activation of the complement system plays a role in target organ damage. The aim of our study was to investigate the influence of cyclosporine (CsA)- induced renal injury on the complement system in the kidney. MATERIALS AND METHODS: Mice fed a low salt (0.01%) diet were treated with vehicle (VH, olive oil, 1mL/kg/day) or CsA (30mg/kg/day) for one or four weeks. Induction of chronic CsA nephrotoxicity was evaluated with renal function and histomorphology. Activation of the complement system was assessed through analysis of the expression of C3, C4d, and membrane attack complex (MAC), and the regulatory proteins, CD46 and CD55. CsA treatment induced renal dysfunction and typical morphology (tubulointerstitial inflammation and fibrosis) at four weeks. RESULTS: CsA-induced renal injury was associated with increased the expression of C3, C4d, and MAC (C9 and upregulation of complement regulatory proteins (CD 46 and CD55). Immunohistochemistry revealed that the activated complement components were mainly confined to the injured tubulointerstitium. CONCLUSION: CsA-induced renal injury is associated with activation of the intrarenal complement system.

Keyword

Cyclosporine; complement; nephrotoxicity; innate immunity

MeSH Terms

Animals
Antigens, CD45/analysis
Antigens, CD46/analysis
Antigens, CD55/analysis
Complement C3/analysis
Complement C4b/analysis
Complement Membrane Attack Complex/analysis
Complement System Proteins/*analysis
Cyclosporine/*toxicity
Disease Models, Animal
Immunity, Innate/drug effects
Immunoblotting
Immunohistochemistry
Immunosuppressive Agents/toxicity
Kidney/*drug effects/immunology/pathology
Kidney Diseases/*chemically induced/immunology
Mice
Microscopy, Confocal
Peptide Fragments/analysis

Figure

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  Fig. 1 Representative photomicrographs of the histomorphology for each group. CsA treatment for four weeks induced interstitial fibrosis. Trichrome stain, original magnification × 200.

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  Fig. 2 Representative photomicrographs of immunohistochemistry for C3. CsA treatment resulted in a significant increase in C3 immunoreactivity. Immunohistochemical staining, original magnification, × 200; insets × 1000.

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  Fig. 3 Representative photomicrographs of immunohistochemistry for C4d. CsA treatment resulted in a significant increase in C4d immunoreactivity (arrow heads). Immunohistochemical staining, original magnification, × 200; insets (a) × 1000.

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  Fig. 4 Double immunofluorescence with vessel markers for α-smooth muscle actin (green) and C4d or C3 (red). Original magnification × 400.

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  Fig. 5 Immunohistochemistry (A) and immunoblotting (B) for membrane attack complex (MAC). Note that the increase in MAC protein expression in renal tubules induced by CsA treatment was confined to the apical side (double arrows) and the tubulointerstitium (asterisks). Magnifications: inset × 1000; others × 200. #p < 0.05 vs. VH.

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  Fig. 6 Immunohistochemistry of CD55. CD55 is strongly expressed in the endothelial cells of the glomeruli and the vascular bundles of the outer medulla (arrows). Dotted lines indicate the vascular bundle of the outer medulla. VB, vascular bundle. Original magnification × 200.

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  Fig. 7 Immunoblotting analysis of CD45. CsA treatment markedly increased the expression of CD45 protein. The relative optical densities (%) are presented with the VH group designated as 100%. #p < 0.05 vs. VH.

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