title: Necrotic renal epithelial cell inhibits renal interstitial fibroblast activation: role of protein tyrosine phosphatase 1B.
abstract: Our recent studies showed that contents of necrotic renal proximal tubular cells (RPTC) from 2 × 10(6) cells/ml directly induced death of cultured renal interstitial fibroblasts. However, it remains unknown whether nonlethal number of necrotic RPTC would also alter the fate of renal interstitial fibroblasts. To address this issue, renal interstitial fibroblasts (NRK-49F) were exposed to necrotic RPTC supernatant (RPTC-Sup) obtained from 2 × 10(4) to 5 × 10(5) cells/ml. These concentrations of RPTC did not induce cell death, but led to inactivation of renal fibroblasts as indicated by reduced expression of α-smooth muscle actin and fibronectin, two hallmarks of activated fibroblasts. Concurrently, the same doses of necrotic RPTC-Sup suppressed phosphorylation of epidermal growth factor receptor (EGFR) and signal transducers and activators of transcription-3 (STAT3) in a time- and dose-dependent manner, but did not affect phosphorylation of platelet-derived growth factor receptor-β, AKT, and extracellular signal-regulated kinase 1/2. The presence of sodium orthovanadate, a general protein tyrosine phosphatase (PTP) inhibitor or TCS-401 (a selective PTP1B inhibitor), abrogated those effects of RPTC-Sup, whereas coincubation with the EGFR inhibitor (Gefitinib) or silencing of EGFR with siRNA preserved the ability of RPTC-Sup in suppressing renal fibroblast activation and STAT3 phosphorylation. Moreover, RPTC-Sup treatment induced PTP1B phosphorylation and its interaction with EGFR. Collectively, these results indicate that nonlethal necrotic RPTC-Sup can induce inactivation of renal interstitial fibroblasts, which occurs through a mechanism involved in PTP1B-mediated inhibition of EGFR signaling.
authors.affiliation: Department of Medicine, Rhode Island Hospital and Alpert Medical School of Brown University, Providence, RI 02903, USA.
authors.forename: Murugavel
authors.initials: M
authors.lastname: Ponnusamy
authors.forename: Li
authors.initials: L
authors.lastname: Ma
authors.forename: Shougang
authors.initials: S
authors.lastname: Zhuang
mesh_terms: D058186:Acute Kidney Injury / Q000201:enzymology* / Q000503:physiopathology; D000818:Animals; D002460:Cell Line; D049109:Cell Proliferation; D066246:ErbB Receptors / Q000378:metabolism; D007668:Kidney / Q000473:pathology*; D007687:Kidney Tubules, Proximal / Q000503:physiopathology*; D051379:Mice; D058628:Myofibroblasts / Q000502:physiology*; D009336:Necrosis; D054562:Protein Tyrosine Phosphatase, Non-Receptor Type 1 / Q000378:metabolism*; D051381:Rats; D058486:Receptors, Purinergic P2X7 / Q000378:metabolism; D050796:STAT3 Transcription Factor / Q000378:metabolism
chemical_list: D058486:Receptors, Purinergic P2X7; D050796:STAT3 Transcription Factor; C494088:Stat3 protein, rat; C483803:Egfr protein, rat; D066246:ErbB Receptors; D054562:Protein Tyrosine Phosphatase, Non-Receptor Type 1; C516610:Ptpn1 protein, rat