NLR Family Pyrin Domain‐Containing 3 Inflammasome Activation in Hepatic Stellate Cells Induces Liver Fibrosis in Mice

作者：María Eugenia Inzaugarat, Casey D. Johnson, Theresa Maria Holtmann, Matthew D. McGeough, Christian Trautwein, Bettina G. Papouchado, Robert F. Schwabe, Hal M. Hoffman, Alexander Wree, Ariel E. Feldstein · 发表于：Hepatology · 年份：2018 · DOI：10.1002/hep.30252 · 被引用次数：146 · 研究领域：Liver Disease Diagnosis and Treatment、Liver physiology and pathology、Liver Diseases and Immunity

The NLR family pyrin domain‐containing 3 (NLRP3) inflammasome plays an important role in liver fibrosis (LF) development. However, the mechanisms involved in NLRP3‐induced fibrosis are unclear. Our aim was to test the hypothesis that the NLRP3 inflammasome in hepatic stellate cells (HSCs) can directly regulate their activation and contribute to LF. Primary HSCs isolated from wild‐type (WT), Nlrp3–/– , or Nlrp3L351PneoR knock‐in crossed to inducible (estrogen receptor Cre‐CreT) mice were incubated with lipopolysaccharide (LPS) and adenosine triphosphate (ATP), or 4OH‐tamoxifen, respectively. HSC‐specific Nlrp3L351P knock‐in mice were generated by crossing transgenic mice expressing lecithin retinol acyltransferase (Lrat)‐driven Cre and maintained on standard rodent chow for 6 months. Mice were then sacrificed; liver tissue and serum were harvested. Nlrp3 inflammasome activation along with HSC phenotype and fibrosis were assessed by RT‐PCR, western blotting, fluorescence‐activated cell sorting (FACS), enzyme‐linked immunosorbent assay, immunofluorescence (IF), and immunohistochemistry (IHC). Stimulated WT HSCs displayed increased levels of NLRP3 inflammasome‐induced reactive oxygen species (ROS) production and cathepsin B activity, accompanied by an up‐regulation of mRNA and protein levels of fibrotic makers, an effect abrogated in Nlrp3–/– HSCs. Nlrp3L351P CreT HSCs also showed elevated mRNA and protein expression of fibrotic markers 24 hours after inflammasome activation indu...