Cannabinoid receptor 2 (CB2) has shown anti-inflammatory and anti-fibrogenic properties by regulating immune cells. However, the relationship between CB1 and inflammatory cells in liver injury is still undefined. Methods: ICR mice were lethally irradiated and received bone marrow (BM) transplantation from enhanced green fluorescent protein transgenic mice. Four Epacadostat research buy weeks later, mice of BM-rebuild were subjected to carbon tetrachloride
(CCl4)-induced liver injury. Boyden chamber was used for cell migration assay. Immunofluorescent staining and FACS were used to identify BM-derived monocyte/macrophage (BMM). Latex beads were used to perform phagocytosis of BMM. Expressions of inflammatory cytokines, CB1 and CB2 etc were determined by Western blot, RT-qPCR and Cytometric Bead Array. Active RhoA and Rac1 were measured by pull-down assay. Results: Endocanna-binoid-related
enzymes and receptors, which correlated find protocol with inflammation/fibrosis parameters, showed significant changes in mouse CCl4-injured liver. BMM significantly expressed CB1 and CB2. In vitro, the treatment of mAEA (CB1 agonist) caused a concentration-dependent increase in BMM migration, but JWH133 (CB2 agonist) had no influence. Pharmacological inhibition or genetic knockdown of CB1 markedly attenuated mAEA-mediated migration, but AM630 (CB2 antagonist) or CB2 knockdown hardly influence mAEA-mediated migration. Pull-down analysis showed that mAEA promoted active GTP-bound RhoA protein levels of BMM but that Rac1-GTP changed slightly. This added GTP-bound RhoA conformation by mAEA was inhibited by AM281 or pertussis toxin (PTX, G(a)i/o protein inhibitor). Moreover, mAEA-mediated migration was impaired by PTX and Y27632 (the inhibitor of Rho-associated protein kinase ROCK, downstream molecule of Rho) pretreatment, suggesting mAEA-mediated migration depending on G(a)i/o/RhoA/ROCK signal
axis. In vivo, blockade of CB1 inhibited the recruitment of BMM, whereas no effects on the migration of BM-originated T cells, dendritic cells and neutrophils. Furthermore, activation of CB1 enhanced the phagocytic activity and cytokines expression of BMM, such as TNF-α, IL-6, and MCP-1. In vivo, the blockade of CB1 markedly down-regulated the mRNA and protein levels of TNF-α, IL-6, IL-10, IL-12, IFN-γ and MCP-1 in injured liver. Notably, inhibition MCE公司 of CB1 also ameliorated hepatic inflammation and fibrosis. Conclusion: CB1 is involved in migration, phagocytosis and inflammatory cytokines production of BMM. The blockade of CB1 significantly attenuates hepatic inflammation and fibrosis. Disclosures: The following people have nothing to disclose: Ping Mai, Le Yang, Lin Wang, Lei Tian, Shuangshuang Jia, Yuanyuan Zhang, Xin Liu, Lin Yang, Liying Li It appears that hepatic progenitor cells may be transformed into myofibroblasts and contribute a profibrotic effect in sustaining the progression towards cirrhosis.