Bacterial DNA Promotes Proliferation of Rat Pancreatic Stellate Cells Thorough Toll-Like Receptor 9

Abstract

Our findings demonstrate that bacterial DNA promotes migration and proliferation of PSCs and suggest that bacterial DNA can initiate and sustain pancreatic inflammation and fibrosis by means of TLR9.

ERK pathway and sheddases play an essential role in ethanol-induced CX3CL1 release in pancreatic stellate cells

“…9 Purity of PSCs of 490% was confirmed by the typical star-like configuration of the cells and by presence of vitamin A autofluorescence. The cells were maintained in complete Dulbecco's modified Eagle medium (DMEM)/F-12, which is a mixture of DMEM and Ham's F-12 nutrient mixture (Wako Pure Chemicals, Osaka, Japan) supplemented with 10% fetal bovine serum, 50 units/ml of penicillin, and 50 mg/ml of streptomycin (Invitrogen, Carlsbad, CA, USA).…”

“…9 PSCs were serum starved and incubated for 24 h at 37 1C and fixed in 4% paraformaldehyde. After blocking with 2% normal bovine serum albumin, cells were incubated with rabbit anti-rat CX3CL1, MMP-2, and ADAM17 antibody at 1:100 dilution and with mouse anti-rat a-SMA antibody at 1:400 dilution overnight at 4 1C.…”

“…9 Cells were lysed in RIPA buffer (Nacalai Tesque, Kyoto, Japan) and cellular proteins (approximately 50 mg) were fractionated by electrophoresis on a 10% sodium dodecyl sulfate polyacrylamide gel (Bio-Rad, Hercules, CA, USA). The proteins were transferred to a nitrocellulose membrane (Bio-Rad), and the membrane was incubated for 2 to 6 h with primary antibodies at 1:1000 dilutions.…”

Pancreatic Stellate Cells and CX3CR1

“…PSCs were isolated from the pancreas of normal male Wistar rats (weighing 180–200 g) by a density-gradient centrifugation method as previously described 56 . Briefly, purity of PSCs was more than 90% as evaluated by the typical star-like configuration and by detecting vitamin A autofluorescence.…”

The role of exosomes in the pathogenesis of pancreatic ductal adenocarcinoma

“…Activated PSCs produce inflammatory mediators, including interleukin (IL) -1, IL-6, IL-8, IL-15, CTGF, TGFβ and RANTES which are all capable of activating other nearby quiescent PSCs via an autocrine loop. (Andoh et al , 2000, Shek et al , 2002 Activated PSC play key roles in the development of a fibrotic pancreatic stroma with the synthesis of extracellular matrix proteins such as collagen and enzymes responsible for matrix remodelling such as matrix metalloproteinases (MMPs) and their inhibitors, tissue inhibitor of metalloproteinases (TIMPs) (Nakamura et al , 2011, Phillips et al , 2003.…”