Effect of alcohol on the progression of pancreatitis through the pancreatic stellate cells activated by ferroptosis of pancreatic acinar cells via the TLR4/NOX4/IL-6 signaling pathway
BA Mudeng
Abulikemu Wulayin
Alimu Maimaiti
Abstract：Objective To investigate the mechanisms of alcohol as a high risk factor in inducing pancreatitis,and to explore the role of toll-like receptor 4(TLR4)inhibitor in alleviating the progression of pancreatitis in rats.Methods A total of 30 male,freshly weaned Sprague-Dawley(SD)rats were randomly divided into 5 groups:Control group,Vehicle group,Alcohol group,Alcohol+LPS(Model)group,TLR4 inhibitor+Model group,with 6 rats in each group.Rat pancreatitis model was created by alcohol consumption and lipopolysaccharide(LPS)induction.Western blot was used to determine the expression levels of glutathione peroxidase 4(GPX4),arachidonate 5-lipoxygenase(ALOX5),alpha-smooth muscle actin(α-SMA),TLR4 and NADPH oxidase 4(NOX4)in pancreatic tissues.Enzyme-linked immunosorbent assay(ELISA)was used to determine the interleukin-6(IL-6),reactive oxygen species(ROS)and malondialdehyde(MDA)levels in pancreatic tissues.H&E staining and histopathological analysis were used to analyze the pancreatic tissue damage in rats.Results Compared with Control group,the expression levels of ferroptosis markers GPx4 and ALOX5,pancreatic stellate cells marker α-SMA,TLR4 and NOX4 were significantly up-regulated in Alcohol group and Alcohol+LPS(Model)group(P＜0.05).IL-6,ROS and MDA levels were significantly higher in Alcohol group and Alcohol+LPS(Model)group than those of Control group(P＜0.05).In Control group and Vehicle group,the pancreatic acinar cells were well arranged,with a regular cellular morphology and absence of obvious swelling or shrinking cells in pancreatic tissues.In Alcohol group and Alcohol+LPS(Model)group,the pancreatic acinar cells shrank,with a decreased number of cells per view and a large number of immune cells(pancreatic stellate cells)infiltrated in the space between acinar cells.The Alcohol+LPS(Model)group had more severe inflammatory injury than the Alcohol group.Compared with Alcohol+LPS(Model)group,the Model+TLR4 inhibitor group partially reversed the above indicators and the degree of pancreatic inflammatory injury(P＜0.05).Conclusion Alcohol induces ferroptosis of pancreatic acinar cells,activates pancreatic stellate cells and promotes the progression of pancreatitis through the TLR4/NOX4/IL-6 signaling pathway.TLR4 inhibitor treatment can alleviate the disease progression.
Keywords：pancreatitisalcoholferroptosisTLR4/NOX4/IL-6 pathwaypancreatic acinar cellspancreatic stellate cells
Publication Date：2025-11-26
Online Publishing Date：2025-12-12(First online date of this platform, not the publication date of the document)
Pages：5( 1782-1786 )