Laboratory Investigation

Kidney International (1992) 42, 327–334; doi:10.1038/ki.1992.293

Tumor necrosis factor alpha increases antifibrinolytic activity of cultured human mesangial cells

Quentin Meulders, Ci-Jiang He, Colette Adida, Marie-Noëlle Peraldi, Wolf-Dieter Schleuning, Jean-Daniel Sraer and Eric Rondeau

Service de Néphrologie and INSERM U 64, Hôpital Tenon, Paris, France and Schering AG, Biochemistry Institute, Berlin, Germany

Correspondence: Doctor Q Meulders, Nephrology Unit, Louvain Medical School, Catholic University of Louvain, Avenue Hippocrate 10, B-1200 Bruxelles, Belgium.

Received 16 May 1991; Revised 3 March 1992; Accepted 5 March 1992.

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Abstract

Tumor necrosis factor alpha increases antifibrinolytic activity of cultured human mesangial cells. Tumor necrosis factoralpha (TNFalpha) is likely to exert a major influence in the pathogenesis of glomerulopathies. Besides its proinflammatory properties, TNFalpha interacts with cell growth and synthesis of components of the fibrinolytic system. In this study, we report the effects of recombinant human TNFalpha on the synthesis of tissue-type plasminogen activator (t-PA) and its inhibitor (PAI-1) by human mesangial cells in culture. We first demonstrate that TNFalpha binds specifically to a single class of high affinity receptors (Kd 5.10-11 M; 1500 receptors/cell). TNFalpha has an antimitogenic effect on human mesangial cells since it decreased DNA synthesis, measured by 3H-thymidine incorporation, in a dose-dependent manner. Release of cytosolic LDH and incorporated 51Cr was not increased by 100 ng/ml TNFalpha as compared with control, indicating that this monokine is not cytotoxic for cultured human mesangial cells. Zymographic analysis and reverse fibrin autography disclosed a 120 kD t-PA-PAI-1 complex and a 50 kD free form of PAI-1 in the supernatants of both unstimulated and TNF-stimulated cells; PAI-1 was released in excess and free t-PA was not observed. TNFalpha (0 to 100 ng/ml) had no effect on t-PA synthesis, but enhanced PAI-1 release in a time- and dose-dependent manner (97% increase of PAI-1 synthesis after a 24 hour incubation). This effect was abolished by cycloheximide, suggesting that protein synthesis was required. Northern blot analysis showed that TNFalpha increased the steady-state PAI-1 mRNA levels in a time-dependent manner, with a maximal effect at two hours. Finally, in contrast to what was reported for rat mesangial cells, we failed to demonstrate a production of TNFalpha in human mesangial cells themselves by immunoradiometric assay, immunocytochemistry and Northern blot analysis using a specific TNFalpha cDNA probe. We conclude that TNFalpha has an antimitogenic activity on human mesangial cells in culture and enhances the synthesis of PAI-1. Whether TNFalpha plays a role in glomerular PAI-1 deposition and persistency of fibrin in vivo remains to be determined.

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