• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of jcinvestThe Journal of Clinical InvestigationCurrent IssueArchiveSubscriptionAbout the Journal
J Clin Invest. Nov 15, 1997; 100(10): 2512–2520.
PMCID: PMC508451

Direct in vivo inhibition of the nuclear cell cycle cascade in experimental mesangial proliferative glomerulonephritis with Roscovitine, a novel cyclin-dependent kinase antagonist.


Glomerular injury is characterized by mesangial cell (MC) proliferation and matrix formation. We sought to determine if reducing the activity of cyclin-dependent kinase 2 (CDK2) with the purine analogue, Roscovitine, decreased MC proliferation in vitro and in vivo. Roscovitine (25 microM) inhibited FCS-induced proliferation (P < 0.0001) in cultured MC. Rats with experimental mesangial proliferative glomerulonephritis (Thy1 model) were divided into two groups. A prevention group received daily intraperitoneal injections of Roscovitine in DMSO (2.8 mg/kg) starting at day 1. A treatment group received daily Roscovitine starting at day 3, when MC proliferation was established. Control Thy1 rats received DMSO alone. MC proliferation (PCNA +/OX7 + double immunostaining) was reduced by > 50% at days 5 and 10 in the Roscovitine prevention group, and at day 5 in the treatment group (P < 0.0001). Early administration of Roscovitine reduced immunostaining for collagen type IV, laminin, and fibronectin at days 5 and 10 (r = 0.984; P < 0.001), which was associated with improved renal function (urinary protein/creatinine, blood urea nitrogen, P < 0.05). We conclude that reducing the activity of CDK2 with Roscovitine in experimental glomerulonephritis decreases cell proliferation and matrix production, resulting in improved renal function, and may be a useful therapeutic intervention in disease characterized by proliferation.

Full Text

The Full Text of this article is available as a PDF (412K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Johnson RJ. The glomerular response to injury: progression or resolution? Kidney Int. 1994 Jun;45(6):1769–1782. [PubMed]
  • Sherr CJ. Cancer cell cycles. Science. 1996 Dec 6;274(5293):1672–1677. [PubMed]
  • Sherr CJ. Mammalian G1 cyclins. Cell. 1993 Jun 18;73(6):1059–1065. [PubMed]
  • Baldin V, Lukas J, Marcote MJ, Pagano M, Draetta G. Cyclin D1 is a nuclear protein required for cell cycle progression in G1. Genes Dev. 1993 May;7(5):812–821. [PubMed]
  • Ando K, Ajchenbaum-Cymbalista F, Griffin JD. Regulation of G1/S transition by cyclins D2 and D3 in hematopoietic cells. Proc Natl Acad Sci U S A. 1993 Oct 15;90(20):9571–9575. [PMC free article] [PubMed]
  • Ohtsubo M, Theodoras AM, Schumacher J, Roberts JM, Pagano M. Human cyclin E, a nuclear protein essential for the G1-to-S phase transition. Mol Cell Biol. 1995 May;15(5):2612–2624. [PMC free article] [PubMed]
  • Girard F, Strausfeld U, Fernandez A, Lamb NJ. Cyclin A is required for the onset of DNA replication in mammalian fibroblasts. Cell. 1991 Dec 20;67(6):1169–1179. [PubMed]
  • Draetta G. Cdc2 activation: the interplay of cyclin binding and Thr161 phosphorylation. Trends Cell Biol. 1993 Sep;3(9):287–289. [PubMed]
  • Tsai LH, Lees E, Faha B, Harlow E, Riabowol K. The cdk2 kinase is required for the G1-to-S transition in mammalian cells. Oncogene. 1993 Jun;8(6):1593–1602. [PubMed]
  • Sherr CJ, Roberts JM. Inhibitors of mammalian G1 cyclin-dependent kinases. Genes Dev. 1995 May 15;9(10):1149–1163. [PubMed]
  • Yamamoto T, Wilson CB. Complement dependence of antibody-induced mesangial cell injury in the rat. J Immunol. 1987 Jun 1;138(11):3758–3765. [PubMed]
  • Yamamoto T, Wilson CB. Quantitative and qualitative studies of antibody-induced mesangial cell damage in the rat. Kidney Int. 1987 Oct;32(4):514–525. [PubMed]
  • Johnson RJ, Raines EW, Floege J, Yoshimura A, Pritzl P, Alpers C, Ross R. Inhibition of mesangial cell proliferation and matrix expansion in glomerulonephritis in the rat by antibody to platelet-derived growth factor. J Exp Med. 1992 May 1;175(5):1413–1416. [PMC free article] [PubMed]
  • Shankland SJ, Hugo C, Coats SR, Nangaku M, Pichler RH, Gordon KL, Pippin J, Roberts JM, Couser WG, Johnson RJ. Changes in cell-cycle protein expression during experimental mesangial proliferative glomerulonephritis. Kidney Int. 1996 Oct;50(4):1230–1239. [PubMed]
  • Shankland SJ, Pippin J, Flanagan M, Coats SR, Nangaku M, Gordon KL, Roberts JM, Couser WG, Johnson RJ. Mesangial cell proliferation mediated by PDGF and bFGF is determined by levels of the cyclin kinase inhibitor p27Kip1. Kidney Int. 1997 Apr;51(4):1088–1099. [PubMed]
  • Meijer L. Chemical inhibitors of cyclin-dependent kinases. Trends Cell Biol. 1996 Oct;6(10):393–397. [PubMed]
  • Meijer L. Chemical inhibitors of cyclin-dependent kinases. Prog Cell Cycle Res. 1995;1:351–363. [PubMed]
  • Veselý J, Havlicek L, Strnad M, Blow JJ, Donella-Deana A, Pinna L, Letham DS, Kato J, Detivaud L, Leclerc S, et al. Inhibition of cyclin-dependent kinases by purine analogues. Eur J Biochem. 1994 Sep 1;224(2):771–786. [PubMed]
  • Jeffrey PD, Russo AA, Polyak K, Gibbs E, Hurwitz J, Massagué J, Pavletich NP. Mechanism of CDK activation revealed by the structure of a cyclinA-CDK2 complex. Nature. 1995 Jul 27;376(6538):313–320. [PubMed]
  • Lees E. Cyclin dependent kinase regulation. Curr Opin Cell Biol. 1995 Dec;7(6):773–780. [PubMed]
  • De Azevedo WF, Leclerc S, Meijer L, Havlicek L, Strnad M, Kim SH. Inhibition of cyclin-dependent kinases by purine analogues: crystal structure of human cdk2 complexed with roscovitine. Eur J Biochem. 1997 Jan 15;243(1-2):518–526. [PubMed]
  • Meijer L, Borgne A, Mulner O, Chong JP, Blow JJ, Inagaki N, Inagaki M, Delcros JG, Moulinoux JP. Biochemical and cellular effects of roscovitine, a potent and selective inhibitor of the cyclin-dependent kinases cdc2, cdk2 and cdk5. Eur J Biochem. 1997 Jan 15;243(1-2):527–536. [PubMed]
  • Johnson RJ, Iida H, Alpers CE, Majesky MW, Schwartz SM, Pritzi P, Gordon K, Gown AM. Expression of smooth muscle cell phenotype by rat mesangial cells in immune complex nephritis. Alpha-smooth muscle actin is a marker of mesangial cell proliferation. J Clin Invest. 1991 Mar;87(3):847–858. [PMC free article] [PubMed]
  • Floege J, Eng E, Young BA, Couser WG, Johnson RJ. Heparin suppresses mesangial cell proliferation and matrix expansion in experimental mesangioproliferative glomerulonephritis. Kidney Int. 1993 Feb;43(2):369–380. [PubMed]
  • Butler AR. The Jaffé reaction. Identification of the coloured species. Clin Chim Acta. 1975 Mar 10;59(2):227–232. [PubMed]
  • Crocker CL. Rapid determination of urea nitrogen in serum or plasma without deproteinization. Am J Med Technol. 1967 Sep-Oct;33(5):361–365. [PubMed]
  • Nelson RA, Jr, Jensen J, Gigli I, Tamura N. Methods for the separation, purification and measurement of nine components of hemolytic complement in guinea-pig serum. Immunochemistry. 1966 Mar;3(2):111–135. [PubMed]
  • Hugo C, Pichler R, Meek R, Gordon K, Kyriakides T, Floege J, Bornstein P, Couser WG, Johnson RJ. Thrombospondin 1 is expressed by proliferating mesangial cells and is up-regulated by PDGF and bFGF in vivo. Kidney Int. 1995 Dec;48(6):1846–1856. [PubMed]
  • Floege J, Johnson RJ, Gordon K, Iida H, Pritzl P, Yoshimura A, Campbell C, Alpers CE, Couser WG. Increased synthesis of extracellular matrix in mesangial proliferative nephritis. Kidney Int. 1991 Sep;40(3):477–488. [PubMed]
  • Baker PJ, Ochi RF, Schulze M, Johnson RJ, Campbell C, Couser WG. Depletion of C6 prevents development of proteinuria in experimental membranous nephropathy in rats. Am J Pathol. 1989 Jul;135(1):185–194. [PMC free article] [PubMed]
  • Morishita R, Gibbons GH, Ellison KE, Nakajima M, von der Leyen H, Zhang L, Kaneda Y, Ogihara T, Dzau VJ. Intimal hyperplasia after vascular injury is inhibited by antisense cdk 2 kinase oligonucleotides. J Clin Invest. 1994 Apr;93(4):1458–1464. [PMC free article] [PubMed]
  • Suzuki J, Isobe M, Morishita R, Aoki M, Horie S, Okubo Y, Kaneda Y, Sawa Y, Matsuda H, Ogihara T, et al. Prevention of graft coronary arteriosclerosis by antisense cdk2 kinase oligonucleotide. Nat Med. 1997 Aug;3(8):900–903. [PubMed]
  • Wagner RW. Gene inhibition using antisense oligodeoxynucleotides. Nature. 1994 Nov 24;372(6504):333–335. [PubMed]
  • Wagner RW. The state of the art in antisense research. Nat Med. 1995 Nov;1(11):1116–1118. [PubMed]
  • Tsuboi Y, Shankland SJ, Grande JP, Walker HJ, Johnson RJ, Dousa TP. Suppression of mesangial proliferative glomerulonephritis development in rats by inhibitors of cAMP phosphodiesterase isozymes types III and IV. J Clin Invest. 1996 Jul 15;98(2):262–270. [PMC free article] [PubMed]
  • Johnson RJ, Pritzl P, Iida H, Alpers CE. Platelet-complement interactions in mesangial proliferative nephritis in the rat. Am J Pathol. 1991 Feb;138(2):313–321. [PMC free article] [PubMed]

Articles from The Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


  • Compound
    PubChem Compound links
  • MedGen
    Related information in MedGen
  • PubMed
    PubMed citations for these articles
  • Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...