An immortalized rat pancreatic stellate cell line RP-2 as a new cell model for evaluating pancreatic fibrosis,inflammation and immunity

查看全文 作　　者：Rong-Li [1]Piao;Ming [1]Xiu;David R [2,3]Brigstock;Run-Ping [1]Gao

机构地区：[1]Department of Hepatic-biliary-pancreatic Medicine,the First Hospital of Jilin University;[2]The Research Institute at Nationwide Children's Hospital;[3]Division of Pediatric Surgery,Department of Surgery,the Ohio State University

出　　处：《Hepatobiliary & Pancreatic Diseases International》索引2015年第14卷第6期,共9页

基　　金：support by grants from the National Natural Science Foundation of China(81070370 and 81270544) 摘　　要：BACKGROUND: Pancreatic stellate cells(PSCs) play a critical role in the pathogenesis of pancreatic fibrosis and have emerging functions as progenitor cells,immune cells or intermediaries in pancreatic exocrine secretion. Increasing evidence has shown that desmin as an exclusive cytoskeleton marker of PSC is only expressed in part of these cells. This study was to establish a desmin-positive PSC cell line and evaluate its actions on pancreatic fibrosis,inflammation and immunity.METHODS: The presence of cytoskeletal proteins,integrin α5β1 or TLR4,was determined by immunocytochemistry while the production of desmin,collagen I,MMP-1,MMP-2,TIMP-2,or CD14 was evaluated by Western blotting. The levels of desmin,collagen I,IL-1 and IL-6 m RNA were determined by real-time quantitative PCR. The secretion of cytokines was detected by ELISA. Cell function was assessed using adhesion,migration,or proliferation assays. RESULTS: A stable activated rat PSC cell line(designated as RP-2) was established by RSV promoter/enhancer-driven SV40 large T antigen expression. RP-2 cells retained typical PSC properties,exhibited a myofibroblast-like phenotype and persistently produced desmin. The cells produced collagen I protein,matrix metalloproteinases and inhibitors thereof. RP-2 cells demonstrated typical PSC functions,including proliferation,adherence,and migration,the latter two of which occurred in response to fibronectin and were mediated byintegrin α5β1. TLR4 and its response genes including proinflammatory cytokines(IL-1,IL-6,TNF-α) and chemotactic cytokines(MCP-1,MIP-1α,Rantes) were produced by RP-2 cells and activated by LPS. LPS-induced IL-1 or IL-6 m RNA expression in this cell line was fully blocked with My D88 inhibitor.CONCLUSION: RP-2 cells provide a novel tool for analyzing the properties and functions of PSCs in the pathogenesis of fibrosis,inflammation and immunity in the pancreas. 关键词：胰腺外分泌细胞模型细胞免疫纤维化炎症永生化评价大鼠

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An immortalized rat pancreatic stellate cell line RP-2 as a new cell model for evaluating pancreatic fibrosis,inflammation and immunity

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