We measured mRNA levels of adrenomedullin (AM), C-type natriuretic peptide (CNP), vascular endothelial growth factor (VEGF), interleukin 1β (IL-1β) and interleukin 6 (IL-6) in peripheral blood mononuclear cells (PBMC) of patients with IgA nephropathy. To evaluate these mRNA levels, we employed a real-time quantitative PCR method which was performed using a hybridization probe labeled with two fluorescence dyes. This strategy was found to afford the standard curves with a high correlation, suggesting that this method is useful for evaluations of mRNA levels. By this method, levels of AM, CNP, VEGF, IL-1β and IL-6 mRNA in PBMC of 49 IgA nephropathy patients and 35 healthy volunteers were evaluated. Among the mRNAs examined, AM mRNA levels were significantly lower in severe-grade than in mild-grade IgA nephropathy patients. Furthermore, AM mRNA levels correlated with CNP mRNA levels in PBMC of patients with IgA nephropathy, and each peptide generated from these mRNAs has antiproliferative effects on mesangial cells. These data indicate that gene expression of AM in PBMC is regulated according to the pathophysiological states of IgA nephropathy and that decreased AM production may contribute to the progression of IgA nephropathy.

1.
Lai KN, Leung JCK, Li PKT, Lui SF: Cytokine production by peripheral blood mononuclear cells in IgA nephropathy. Clin Exp Immunol 1991;85:240–245.
2.
Schena FP, Mastrolitti G, Jirillo E, Munno I, Pellegrino N, Fracasso AR, Aventaggiato L: Increased production of interleukin-2 and IL-2 receptor in primary IgA nephropathy. Kidney Int 1989;35:875–879.
3.
De Caestecker MP, Bottomley M, Telfer BA, Hutchinson IV, Vose BM, Ballardie FW: Detection of abnormal peripheral blood mononuclear cell cytokine networks in human IgA nephropathy. Kidney Int 1993;44:1298–1308.
4.
Matsumoto K: IL-8 release from cultured peripheral blood monocytes of patients with glomerulonephritis. Clin Exp Immunol 1995;99:106–111.
5.
Ichinose H, Miyazaki M, Koji T, Furusu A, Ozono Y, Harada T, Shin M: Detection of cytokine mRNA-expressing cells in peripheral blood of patients with IgA nephropathy using non-radioactive in situ hybridization. Clin Exp Immunol 1996;103:125–132.
6.
Matsumoto K: Spontaneous and lipopolysaccharide-stimulated secretion of cytokines by peripheral blood monocytes in IgA nephropathy is inhibited by interleukin-10. Nephron 1996;73:305–309.
7.
Matsumoto K, Ohi H, Kanmatsuse K: Interleukin 10 and interleukin 13 synergize to inhibit vascular permeability factor release by peripheral blood mononuclear cells from patients with lipoid nephrosis. Nephron 1997;77:212–218.
8.
Chini EN, Choi E, Grande JP, Burnet JC, Dousa TP: Adrenomedullin suppresses mitogenesis in rat mesangial cells via cAMP pathway. Biochem Biophys Res Commun 1995;215:868–873.
9.
Chini EN, Chini CCS, Bolliger C, Jougasaki M, Grande JP, Burnett JC, Dousa TP: Cytoprotective effects of adrenomedullin in glomerular cell injury: Central role of cAMP signaling pathway. Kidney Int 1997;52:917–925.
10.
Canaan K, Ostendorf T, Zander K, Koch KM, Floege J: C-type natriuretic peptide inhibits mesangial cell proliferation and matrix accumulation in vivo. Kidney Int 1998;53:1143–1151.
11.
Kubo A, Iwano M, Minamino N, Sato H, Nishino T, Hirata E, Akai Y, Shiiki H, Kitamura K, Kangawa K, Matsuo H, Dohi K: Measurement of plasma and urinary adrenomedullin in patients with IgA nephropathy. Nephron 1998;78:389–394.
12.
Iijima K, Yoshikawa N, Connolly DT, Nakamura H: Human mesangial cells and peripheral blood mononuclear cells produce vascular permeability factor. Kidney Int 1993;44:959–966.
13.
Iruela-Arispe L, Gordon K, Hugo C, Duijvestijn AM, Claffey KP, Reilly M, Couser WG, Alpers CE, Johnson RJ: Participation of glomerular endothelial cells in the capillary repair of glomerulonephritis. Am J Pathol 1995;147:1715–1727.
14.
Heid CA, Stevens J, Livak KJ, Williams PM: Real time quantitative PCR. Genome Res 1996;6:986–994.
15.
Gibson UEM, Heid CA, Williams PM: A novel method for real time quantitative RT-PCR. Genome Res 1996;6:995–1001.
16.
Kubo A, Minamino N, Isumi Y, Katafuchi T, Kangawa K, Dohi K, Matsuo H: Production of adrenomedullin in macrophage cell line and peritoneal macrophage. J Biol Chem 1998;273:16730–16738.
17.
Andreoli SP, Bergstein JM:Treatment of severe IgA nephropathy in children. Pediatr Nephrol 1989;3:248–253.
18.
Kitamura K, Sakata J, Kangawa K, Kojima M, Matsuo H, Eto T: Cloning and characterization of cDNA encoding a precursor for human adrenomedullin. Biochem Biophys Res Commun 1993;194:720–725.
19.
Kojima M, Minamino N, Kangawa K, Matsuo H: Cloning and sequence analysis of a cDNA encoding a precursor for rat C-type natriuretic peptide (CNP). FEBS Lett 1990;276:209–213.
20.
Leung DW, Cachianes G, Kuang WJ, Goeddel DV, Ferrara N: Vascular endothelial growth factor is a secreted angiogenic mitogen. Science 1989;246:1306–1309.
21.
Auron PE, Webb AC, Rosenwasser LJ, Mucci SF, Rich A, Wolf SM, Dinarello CA: Nucleotide sequence of human monocyte interleukin 1 precursor cDNA. Proc Natl Acad Sci USA 1984;81:7907–7911.
22.
May LT, Helfgott DC, Sehgal PB: Anti-beta-interferon antibodies inhibit the increased expression of HLA-B7 mRNA in tumor necrosis factor-treated human fibroblasts. Proc Natl Acad Sci USA 1986;83:8957–8961.
23.
Ponte P, Ng SY, Engel J, Gunning P, Kedes L: Evolutionary conservation in the untranslated region of actin mRNAs: DNA sequence of a human beta-actin cDNA. Nucleic Acids Res 1984;12:1687–1696.
24.
Kubo A, Minamino N, Isumi Y, Kangawa K, Dohi K, Matsuo H: Adrenomedullin production is correlated with differentiation in human leukemia cell lines and peripheral blood monocytes. FEBS Lett 1998;426:233–237.
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
You do not currently have access to this content.