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Signal transduction of MCP-1 expression induced by pancreatitis-associated ascitic fluid in pancreatic acinar cells.

Ramudo L, Yubero S, Manso MA, Vicente S, De Dios I - J. Cell. Mol. Med. (2008)

Bottom Line: In response to PAAF, overexpression of MCP-1, phosphorylation of p38-MAPK, degradation of IkappaB alpha and increases in p65 nuclear levels and STAT3 activity were found in acinar cells.MAPK activation was only inhibited by NAC, NF-kappaB activation was repressed by Dx and NAC, and STAT3 pathway was strongly blocked by Dx and significantly reduced by NAC.In conclusion, acinar cells were activated by PAAF to produce MCP-1, mainly via NF-kappaB and STAT3 pathways.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology and Pharmacology, University of Salamanca, Salamanca, Spain.

ABSTRACT
Pancreatitis-associated ascitic fluid (PAAF) is known to contribute to the progression of acute pancreatitis (AP). We have investigated the capability of PAAF to activate the expression of MCP-1 in pancreatic acinar cells and the involvement of MAPK, NF-kappaB and STAT3 as downstream signalling transduction pathways. The actions of dexamethasone (Dx) and N-acetylcysteine (NAC) on the PAAF's acinar effects have also been evaluated. Acinar cells were incubated for 1 hr with PAAF collected from rats with severe AP induced by sodium taurocholate in the absence or presence of Dx (10(-7) M) or NAC (30 mM). MCP-1 mRNA expression, phospho-p38-MAPK, IkappaB alpha, nuclear p65 levels and nuclear translocation of STAT3 were analysed. In response to PAAF, overexpression of MCP-1, phosphorylation of p38-MAPK, degradation of IkappaB alpha and increases in p65 nuclear levels and STAT3 activity were found in acinar cells. PAAF-mediated MCP-1 up-regulation was completely suppressed by Dx and NAC. MAPK activation was only inhibited by NAC, NF-kappaB activation was repressed by Dx and NAC, and STAT3 pathway was strongly blocked by Dx and significantly reduced by NAC. In conclusion, acinar cells were activated by PAAF to produce MCP-1, mainly via NF-kappaB and STAT3 pathways. Both downstream pathways were targeted by Dx and NAC to repress the PAAF-mediated acinar MCP-1 up-regulation.

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Activation of NF-κB in pancreatic acinar cells under basal conditions and under stimulation with pancreatitis-associated ascitic fluid (PAAF, 20% v/v) in the absence or presence of dexamethasone (Dx, 10−7 M) and N-acetylcysteine (NAC, 30 mM). (A) Representative Western blot of IκBα and the mean values of five experiments are shown. Band intensity was determined by densitometry. (B) p65 nuclear levels (NF-κB-DNA binding). Results are mean ± S.E.M. ANOVA followed by Dunnett’s test showed significant differences versus non-PAAF-stimulated cells (basal conditions) (**P < 0.01) and versus PAAF-stimulated cells (♦♦P < 0.01).
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fig04: Activation of NF-κB in pancreatic acinar cells under basal conditions and under stimulation with pancreatitis-associated ascitic fluid (PAAF, 20% v/v) in the absence or presence of dexamethasone (Dx, 10−7 M) and N-acetylcysteine (NAC, 30 mM). (A) Representative Western blot of IκBα and the mean values of five experiments are shown. Band intensity was determined by densitometry. (B) p65 nuclear levels (NF-κB-DNA binding). Results are mean ± S.E.M. ANOVA followed by Dunnett’s test showed significant differences versus non-PAAF-stimulated cells (basal conditions) (**P < 0.01) and versus PAAF-stimulated cells (♦♦P < 0.01).

Mentions: The activation of NF-κB was evaluated by analysis of IκBα (Fig. 4A) and p65 (Fig. 4B). Dx and NAC did not show any effect in acinar cells cultured under basal conditions. PAAF significantly activated NF-κB in acinar cells as indicated by the significant (P < 0.01) degradation of IκBα and the significant (P < 0.01) increase in p65 levels. Both Dx and NAC significantly inhibited NF-κB activation induced by PAAF, by preventing the degradation of IκBα and maintaining p65 values at similar levels to those that were found in acinar cells cultured in the absence of PAAF (basal conditions).


Signal transduction of MCP-1 expression induced by pancreatitis-associated ascitic fluid in pancreatic acinar cells.

Ramudo L, Yubero S, Manso MA, Vicente S, De Dios I - J. Cell. Mol. Med. (2008)

Activation of NF-κB in pancreatic acinar cells under basal conditions and under stimulation with pancreatitis-associated ascitic fluid (PAAF, 20% v/v) in the absence or presence of dexamethasone (Dx, 10−7 M) and N-acetylcysteine (NAC, 30 mM). (A) Representative Western blot of IκBα and the mean values of five experiments are shown. Band intensity was determined by densitometry. (B) p65 nuclear levels (NF-κB-DNA binding). Results are mean ± S.E.M. ANOVA followed by Dunnett’s test showed significant differences versus non-PAAF-stimulated cells (basal conditions) (**P < 0.01) and versus PAAF-stimulated cells (♦♦P < 0.01).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4496145&req=5

fig04: Activation of NF-κB in pancreatic acinar cells under basal conditions and under stimulation with pancreatitis-associated ascitic fluid (PAAF, 20% v/v) in the absence or presence of dexamethasone (Dx, 10−7 M) and N-acetylcysteine (NAC, 30 mM). (A) Representative Western blot of IκBα and the mean values of five experiments are shown. Band intensity was determined by densitometry. (B) p65 nuclear levels (NF-κB-DNA binding). Results are mean ± S.E.M. ANOVA followed by Dunnett’s test showed significant differences versus non-PAAF-stimulated cells (basal conditions) (**P < 0.01) and versus PAAF-stimulated cells (♦♦P < 0.01).
Mentions: The activation of NF-κB was evaluated by analysis of IκBα (Fig. 4A) and p65 (Fig. 4B). Dx and NAC did not show any effect in acinar cells cultured under basal conditions. PAAF significantly activated NF-κB in acinar cells as indicated by the significant (P < 0.01) degradation of IκBα and the significant (P < 0.01) increase in p65 levels. Both Dx and NAC significantly inhibited NF-κB activation induced by PAAF, by preventing the degradation of IκBα and maintaining p65 values at similar levels to those that were found in acinar cells cultured in the absence of PAAF (basal conditions).

Bottom Line: In response to PAAF, overexpression of MCP-1, phosphorylation of p38-MAPK, degradation of IkappaB alpha and increases in p65 nuclear levels and STAT3 activity were found in acinar cells.MAPK activation was only inhibited by NAC, NF-kappaB activation was repressed by Dx and NAC, and STAT3 pathway was strongly blocked by Dx and significantly reduced by NAC.In conclusion, acinar cells were activated by PAAF to produce MCP-1, mainly via NF-kappaB and STAT3 pathways.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology and Pharmacology, University of Salamanca, Salamanca, Spain.

ABSTRACT
Pancreatitis-associated ascitic fluid (PAAF) is known to contribute to the progression of acute pancreatitis (AP). We have investigated the capability of PAAF to activate the expression of MCP-1 in pancreatic acinar cells and the involvement of MAPK, NF-kappaB and STAT3 as downstream signalling transduction pathways. The actions of dexamethasone (Dx) and N-acetylcysteine (NAC) on the PAAF's acinar effects have also been evaluated. Acinar cells were incubated for 1 hr with PAAF collected from rats with severe AP induced by sodium taurocholate in the absence or presence of Dx (10(-7) M) or NAC (30 mM). MCP-1 mRNA expression, phospho-p38-MAPK, IkappaB alpha, nuclear p65 levels and nuclear translocation of STAT3 were analysed. In response to PAAF, overexpression of MCP-1, phosphorylation of p38-MAPK, degradation of IkappaB alpha and increases in p65 nuclear levels and STAT3 activity were found in acinar cells. PAAF-mediated MCP-1 up-regulation was completely suppressed by Dx and NAC. MAPK activation was only inhibited by NAC, NF-kappaB activation was repressed by Dx and NAC, and STAT3 pathway was strongly blocked by Dx and significantly reduced by NAC. In conclusion, acinar cells were activated by PAAF to produce MCP-1, mainly via NF-kappaB and STAT3 pathways. Both downstream pathways were targeted by Dx and NAC to repress the PAAF-mediated acinar MCP-1 up-regulation.

Show MeSH
Related in: MedlinePlus