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Protective effect of resin adsorption on septic plasma-induced tubular injury.

Cantaluppi V, Weber V, Lauritano C, Figliolini F, Beltramo S, Biancone L, De Cal M, Cruz D, Ronco C, Segoloni GP, Tetta C, Camussi G - Crit Care (2010)

Bottom Line: Plasma adsorption significantly decreased these effects and abated the concentrations of several soluble mediators.The alteration of cell polarity, morphogenesis, protein reabsorption and the down-regulation of the tight junction molecule ZO-1, of the sodium transporter NHE3, of the glucose transporter GLUT-2 and of the endocytic receptor megalin all induced by septic plasma were significantly reduced by resin adsorption.All these biological effects are related to the presence of circulating inflammatory mediators that can be efficiently removed by resin adsorption with a consequent limitation of tubular cell injury.

View Article: PubMed Central - HTML - PubMed

Affiliation: Center for Experimental Medical Research (CeRMS), University of Torino, Via Santena 5, Torino 10126, Italy. vincenzo.cantaluppi@unito.it

ABSTRACT

Introduction: A pro-apoptotic effect of circulating mediators on renal tubular epithelial cells has been involved in the pathogenesis of sepsis-associated acute kidney injury (AKI). Adsorption techniques have been showed to efficiently remove inflammatory cytokines from plasma. The aim of this study was to evaluate the efficiency of the hydrophobic resin Amberchrom CG161 M to adsorb from septic plasma soluble mediators involved in tubular injury.

Methods: We enrolled in the study 10 critically ill patients with sepsis-associated AKI and we evaluated the effects of their plasma on granulocyte adhesion, apoptosis and functional alterations of cultured human kidney tubular epithelial cells. We established an in vitro model of plasma adsorption and we studied the protective effect of unselective removal of soluble mediators by the Amberchrom CG161 M resin on septic plasma-induced tubular cell injury.

Results: Plasma from septic patients induced granulocyte adhesion, apoptosis and altered polarity in tubular cells. Plasma adsorption significantly decreased these effects and abated the concentrations of several soluble mediators. The inhibition of granulocyte adhesion to tubular cells was associated with the down-regulation of ICAM-1 and CD40. Resin adsorption inhibited tubular cell apoptosis induced by septic plasma by down-regulating the activation of caspase-3, 8, 9 and of Fas/death receptor-mediated signalling pathways. The alteration of cell polarity, morphogenesis, protein reabsorption and the down-regulation of the tight junction molecule ZO-1, of the sodium transporter NHE3, of the glucose transporter GLUT-2 and of the endocytic receptor megalin all induced by septic plasma were significantly reduced by resin adsorption.

Conclusions: Septic plasma induced a direct injury of tubular cells by favouring granulocyte adhesion, by inducing cell apoptosis and by altering cell polarity and function. All these biological effects are related to the presence of circulating inflammatory mediators that can be efficiently removed by resin adsorption with a consequent limitation of tubular cell injury.

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Effect of resin adsorption on albumin internalization and expression of megalin by TEC. Representative FACS and confocal microscopy analysis of FITC-albumin uptake (green fluorescence) and megalin expression (red fluorescence) in tubular epithelial cells (TEC) incubated with control healthy plasma or septic plasma before and after (Septic + CG161 M) Amberchrom resin adsorption. For FACS analysis, Kolomogorov Smirnov statistical analysis was performed. In merge images, nuclei were counterstained by 0.5 μg/ml Hoechst.
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Figure 8: Effect of resin adsorption on albumin internalization and expression of megalin by TEC. Representative FACS and confocal microscopy analysis of FITC-albumin uptake (green fluorescence) and megalin expression (red fluorescence) in tubular epithelial cells (TEC) incubated with control healthy plasma or septic plasma before and after (Septic + CG161 M) Amberchrom resin adsorption. For FACS analysis, Kolomogorov Smirnov statistical analysis was performed. In merge images, nuclei were counterstained by 0.5 μg/ml Hoechst.

Mentions: Septic plasma significantly reduced TER, an indicator of TEC polarity. This effect was abrogated in the presence of Amberchrom resin-treated plasma (Figure 6a). Further evidence for the maintenance of TEC polarity and function came from the observation that Amberchrom resin abrogated the down-regulation of the tight junction protein ZO-1, proximal tubular cell sodium transporter NHE3 and glucose transporter GLUT-2, which were all induced by septic plasma (Figure 6b). In addition, the reduced adhesion of TEC to the extracellular matrixes fibronectin/type IV collagen and Matrigel observed in the presence of septic plasma was significantly inhibited after Amberchrom resin adsorption (Figure 7a). TEC cultured on Matrigel-coated plates showed a typical morphology characterized by early scattering and branching morphogenesis that was reduced after incubation with septic plasma (Figure 7b). In contrast, TEC morphogenesis was not affected by incubation with Amberchrom-adsorbed plasma (Figure 7b). Moreover, we found that septic plasma induced the down-regulation of the endocytic receptor megalin, a molecule involved in tubular re-adsorption of filtered proteins (Figure 8). The decreased expression of megalin was not observed in the presence of Amberchrom resin-treated plasma (Figure 8). This phenomenon was probably responsible for the preserved ability of TEC to internalize FITC-labeled albumin (Figure 8).


Protective effect of resin adsorption on septic plasma-induced tubular injury.

Cantaluppi V, Weber V, Lauritano C, Figliolini F, Beltramo S, Biancone L, De Cal M, Cruz D, Ronco C, Segoloni GP, Tetta C, Camussi G - Crit Care (2010)

Effect of resin adsorption on albumin internalization and expression of megalin by TEC. Representative FACS and confocal microscopy analysis of FITC-albumin uptake (green fluorescence) and megalin expression (red fluorescence) in tubular epithelial cells (TEC) incubated with control healthy plasma or septic plasma before and after (Septic + CG161 M) Amberchrom resin adsorption. For FACS analysis, Kolomogorov Smirnov statistical analysis was performed. In merge images, nuclei were counterstained by 0.5 μg/ml Hoechst.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2875506&req=5

Figure 8: Effect of resin adsorption on albumin internalization and expression of megalin by TEC. Representative FACS and confocal microscopy analysis of FITC-albumin uptake (green fluorescence) and megalin expression (red fluorescence) in tubular epithelial cells (TEC) incubated with control healthy plasma or septic plasma before and after (Septic + CG161 M) Amberchrom resin adsorption. For FACS analysis, Kolomogorov Smirnov statistical analysis was performed. In merge images, nuclei were counterstained by 0.5 μg/ml Hoechst.
Mentions: Septic plasma significantly reduced TER, an indicator of TEC polarity. This effect was abrogated in the presence of Amberchrom resin-treated plasma (Figure 6a). Further evidence for the maintenance of TEC polarity and function came from the observation that Amberchrom resin abrogated the down-regulation of the tight junction protein ZO-1, proximal tubular cell sodium transporter NHE3 and glucose transporter GLUT-2, which were all induced by septic plasma (Figure 6b). In addition, the reduced adhesion of TEC to the extracellular matrixes fibronectin/type IV collagen and Matrigel observed in the presence of septic plasma was significantly inhibited after Amberchrom resin adsorption (Figure 7a). TEC cultured on Matrigel-coated plates showed a typical morphology characterized by early scattering and branching morphogenesis that was reduced after incubation with septic plasma (Figure 7b). In contrast, TEC morphogenesis was not affected by incubation with Amberchrom-adsorbed plasma (Figure 7b). Moreover, we found that septic plasma induced the down-regulation of the endocytic receptor megalin, a molecule involved in tubular re-adsorption of filtered proteins (Figure 8). The decreased expression of megalin was not observed in the presence of Amberchrom resin-treated plasma (Figure 8). This phenomenon was probably responsible for the preserved ability of TEC to internalize FITC-labeled albumin (Figure 8).

Bottom Line: Plasma adsorption significantly decreased these effects and abated the concentrations of several soluble mediators.The alteration of cell polarity, morphogenesis, protein reabsorption and the down-regulation of the tight junction molecule ZO-1, of the sodium transporter NHE3, of the glucose transporter GLUT-2 and of the endocytic receptor megalin all induced by septic plasma were significantly reduced by resin adsorption.All these biological effects are related to the presence of circulating inflammatory mediators that can be efficiently removed by resin adsorption with a consequent limitation of tubular cell injury.

View Article: PubMed Central - HTML - PubMed

Affiliation: Center for Experimental Medical Research (CeRMS), University of Torino, Via Santena 5, Torino 10126, Italy. vincenzo.cantaluppi@unito.it

ABSTRACT

Introduction: A pro-apoptotic effect of circulating mediators on renal tubular epithelial cells has been involved in the pathogenesis of sepsis-associated acute kidney injury (AKI). Adsorption techniques have been showed to efficiently remove inflammatory cytokines from plasma. The aim of this study was to evaluate the efficiency of the hydrophobic resin Amberchrom CG161 M to adsorb from septic plasma soluble mediators involved in tubular injury.

Methods: We enrolled in the study 10 critically ill patients with sepsis-associated AKI and we evaluated the effects of their plasma on granulocyte adhesion, apoptosis and functional alterations of cultured human kidney tubular epithelial cells. We established an in vitro model of plasma adsorption and we studied the protective effect of unselective removal of soluble mediators by the Amberchrom CG161 M resin on septic plasma-induced tubular cell injury.

Results: Plasma from septic patients induced granulocyte adhesion, apoptosis and altered polarity in tubular cells. Plasma adsorption significantly decreased these effects and abated the concentrations of several soluble mediators. The inhibition of granulocyte adhesion to tubular cells was associated with the down-regulation of ICAM-1 and CD40. Resin adsorption inhibited tubular cell apoptosis induced by septic plasma by down-regulating the activation of caspase-3, 8, 9 and of Fas/death receptor-mediated signalling pathways. The alteration of cell polarity, morphogenesis, protein reabsorption and the down-regulation of the tight junction molecule ZO-1, of the sodium transporter NHE3, of the glucose transporter GLUT-2 and of the endocytic receptor megalin all induced by septic plasma were significantly reduced by resin adsorption.

Conclusions: Septic plasma induced a direct injury of tubular cells by favouring granulocyte adhesion, by inducing cell apoptosis and by altering cell polarity and function. All these biological effects are related to the presence of circulating inflammatory mediators that can be efficiently removed by resin adsorption with a consequent limitation of tubular cell injury.

Show MeSH
Related in: MedlinePlus