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Abstract: SA-PO580

PMMA-Based Continuous-Adsorbitive-Hemofiltration Abrogated Complement Activation and Tubular Senescence-Associated Secretory Phenotype (SASP) in LPS-Induced AKI

Session Information

Category: Acute Kidney Injury

  • 103 AKI: Mechanisms

Authors

  • Castellano, Giuseppe, University of Bari, BARI, Italy
  • Stasi, Alessandra, University of Bari, BARI, Italy
  • Divella, Chiara, University of Bari, BARI, Italy
  • Franzin, Rossana, University of Bari, BARI, Italy
  • Sallustio, Fabio, University of Bari, BARI, Italy
  • Curci, Claudia, University of Bari, BARI, Italy
  • Picerno, Angela, University of Bari "Aldo Moro", Bari, Italy
  • Ronco, Claudio, Ospedale "S. Bortolo" AZ. ULSS 6, Vicenza, Italy
  • Cantaluppi, Vincenzo, University of Piemonte Orientale (UPO), Novara, Italy
  • de Cal, Massimo, San Bortolo Hospital, Vicenza, Italy
  • Neri, Mauro, San Bortolo Hospital, Vicenza, Italy
  • Lorenzin, Anna, IRRIV, Vicenza, Italy
  • Pertosa, Giovanni B., University of Bari, BARI, Italy
  • Gesualdo, Loreto, University of Bari, BARI, Italy
Background

During sepsis-induced AKI, inflammatory mediators could induce a maladaptive response in tubular epithelial cells (TEC). Hemofeel (TORAY, Japan) is a PMMA membrane based hemofilter indicated for blood purification therapies in continuous modality (CRRT), combining filtration and adsorption.

Methods

After 3h from LPS infusion, 8 pigs (female, 40-50 Kg) were treated with PMMA-CVVH (CH-1.8 W series), working in post-dilution modality (time =7 hours; Qb= 150 ml/min; Effluent rate =2000 ml/h; Substitution Rate = 2000 ml/h; UF rate = 0 ml/h). 8 control pigs received no treatment and developed endotoxemia-induced oliguric AKI. Renal biopsies were collected at 24h and analyzed by IHC and IF. Systemic Complement activation was evaluated by Wieslab kit. In vitro, tubular epithelial cells (TEC) were stimulated with LPS (4µg/ml) for 24h and then analyzed for SASP markers (p53, p21, cyp1b1, Klotho) by qPCR.

Results

A significant activation of Complement classical and alternative pathways was observed at 3h and 24h after LPS infusion. Histological and Confocal analysis revealed extensive Pentraxin-3 and collagen deposits at the interstitial level, along capillaries with diffuse glomerular thrombi compared to healthy pigs (T24LPS vs T24CTR, p <0.05). Interestingly, we found significant TEC senescence with increased p16INK4a expression (IHC T24LPS vs T24CTR p <0.05). PMMA-CVVH treatment significantly reduced Complement activation and antagonized SASP by restoring p16INK4a basal expression (T24LPS vs T24PMMA p <0.05). In accordance, LPS stimulation of TEC (24h) in vitro induced accelerated senescence by up-regulating p53, p21, cyp1b1 (2-DDCT: LPS: 4.3 ± 0.63 vs Basal: 1 ± 0.00, p <0.05) and reducing Klotho expression (p <0.05).

Conclusion

Endotoxemia can induce AKI with systemic Complement activation leading to the development of SASP phenotype in TEC. PMMA treatment might be a new therapeutic approach to prevent Complement activation in LPS-induced AKI.