P2X7R Inhibitor Suppressed Inflammasome-Mediated Hepatic Stellate Cell Activation

BACKGROUND&AIMS:The ATP-gated P2X7receptor (P2X7r) is a promising therapeutic target in fibrosis diseases including liver fibrosis. Although Hpopolysaccharide (LPS) and pro-inflammatory cytokines related to the liver fibrosis, the underlying molecular mechanisms remain to be elucidated.METHODS:The supernatant from LPS-stimulated RAW264.7mouse macrophage adds human hepatic stellate cell (LX-2) for24h. LX-2primed with LPS and subsequently stimulated for30min with3mM of ATP. The P2X7r inhibitor (A438079,10μM) was applied10min before and during ATP.RESULTS:When LX-2were cultured in the presence of LPS, mRNA expression of a-SMA and collagen-I were increased, as well as IL-1β, IL-18and IL-6. And LPS also increased mRNA expression of caspase-1, P2X7r and ASC that the apoptosis-associated speck like CARD-domain containing protein. These mRNA expressions were higher in conditioned medium of RAW264.7cells. The expressions of capase-1and P2X7r were increased by the treatment of conditioned medium of RAW264.7cells with lipopolysaccharide, which also induced the expression of different proinflammatory cytokines, including IL-1β, IL-18and IL-6. Our results suggest P2X7r activation may play a novel and direct role in HSC activation through release of proinflammatory cytokines of its proinflammatory actions via cytokines. Also, when treated the LPS-primed cell with ATP and A438079, ATP-mediated receptor has support for the decline of the cytokines expression.CONCLUSION:P2X7receptor is able to regulate the secretion of IL-1β and have a more close connection with Kupffer cells, may the cytokines stimulated from the Kupffer cell, and then effect the mRNA expression of HSCs as a serial function contribute to fibrosis progression. P2X7r blockades the protective effects via A438079exhibit, attenuating LPS-induced liver injury and fibrosis. Also, P2X7receptor can improve the mature of pro-ILp.