Effect of pioglitazone on inflammatory signaling pathways induced by pancreatitis-associated ascitic fluid in human monocytic cell line THP-1

Abstract

Objective To investigate the inhibitory effect of pioglitazone on inflammatory signaling pathways induced by human pancreatitis-associated ascitic fluid(PAAF) in human monocytic cell line THP-1.Methods PAAF was collected from patients with severe acute pancreatitis.The cultured THP-1 cells were divided into 4 groups: control group(C),PAAF group(A),PPARγ agonist pioglitazone group(P),and PPARγ antagonist GW9662 group(G).Cells in group A were stimulated with PAAF,those in group P were treated with 20 μmol/L PPARγ agonist pioglitazone 2 h after stimulation with PAAF,and those in group G were treated with PPARγ antagonist GW9662,pioglitazone(30 min later) and PAAF(2 h later) in turn.After incubation for 12 h,TNF-α and IL-6 mRNA expression was measured by real-time PCR,and Western blotting analysis was used to examine the expression of the phospho-p38-mitogen-activated protein kinase(MAPK),nuclear transcription factor(NF-κB) p65 levels,and IκB. Results Compared with group C,TNF-α and IL-6 mRNA expression in group A was increased,NF-κB and p38MAPK protein levels were increased,and IκB protein level was decreased(P0.05).Compared with group A,TNF-α and IL-6 mRNA expression was decreased in group P,NF-κB and p38MAPK protein levels were down-regulated,and IκB protein level was increased(P0.05).Compared with group P,TNF-α and IL-6 mRNA expression was increased in group G,NF-κB and p38MAPK protein levels were increased,and IκB protein level was decreased(P0.05).Conclusion The results show that pioglitazone can inhibit PAAF-stimulated THP-1 inflammation by blocking p38MAPK and NF-κB signaling pathway.