13 Although NOX plays an important role in HSC activation and hepatic fibrosis, how various NOX homologues expressed in different hepatic cell types contribute to hepatic fibrogenesis MI-503 chemical structure is unknown. The purpose of the present study was to investigate the contributory role of NOX1 and NOX2 in hepatic fibrosis. We evaluated the effect of genetic NOX1 and NOX2 inactivation on hepatic fibrosis in two different models of experimental fibrogenesis. We also identified NOX1- and NOX2-expressing cell types in the liver. The functional contribution of NOX1 and NOX2 in endogenous liver cells, including HSCs, and in bone marrow (BM)-derived

cells, including Kupffer cells (KCs), to hepatic fibrosis was assessed using either NOX1 or NOX2 BM chimeric mice. Our data indicate that both NOX1 and NOX2 have an important role in hepatic fibrosis in endogenous liver cells, whereas NOX2 has a lesser role in BM-derived cells. These findings may provide new insight into the development of antifibrotic therapy this website targeting nonphagocytic NOX signaling

in the liver without suppression of NOX2-mediated host defense mechanism. α-SMA, α-smooth muscle actin; ALT, alanine aminotransferase; Ang II, angiotensin II; BDL, bile duct ligation; BM, bone marrow; BMT, bone marrow transplantation; CCl4, carbon tetrachloride; CM-H2DCFDA, 2′7′-dichlorofluorescein diacetate; DMEM, Dulbecco’s modified Eagle’s medium; HSC, hepatic stellate cell; KC, Kupffer cell; KO, knockout; mRNA, messenger RNA; NOX, nicotinamide adenine

dinucleotide phosphate oxidase; PCR, polymerase chain reaction; ROS, reactive oxygen species; SEC, sinusoid endothelial cell; TGF-β, transforming growth factor β; WT, wild-type. NOX2 knockout (NOX2KO) mice15 with a C57BL/6 background, which lack a catalytic subunit of phagocytic NOX complex, and WT C57BL/6 control mice were purchased from The Jackson Laboratory (Bar Harbor, MA). NOX1 knockout (NOX1KO) mice16 with a C57BL/6 background were a kind gift from John Engelhardt of the University of Iowa. Eight- to 10-week-old male mice were used. Liver fibrosis was induced either by way of intraperitoneal injection of hepatotoxin carbon tetrachloride (CCl4) or by way of BDL. CCl4 (Sigma Aldrich, St. Louis, MO; diluted 上海皓元医药股份有限公司 1:3 in corn oil) or a vehicle (corn oil) was injected on every third day at a concentration of 2 μL/g body weight 12 times per day. BDL was performed as described, and a sham operation group was used as a control.17 Animals were sacrificed 72 hours after the last CCl4 injection or 3 weeks after BDL and blood and liver samples were collected. All animal studies were approved by The University of California, San Diego Institutional Animal Care and Use Committee (protocol number: S-07088). We performed BM transplantation (BMT) experiments as described with slight modifications.18-21 We flushed the tibias and femurs of donor mice to obtain BM.