PMID- 26764184 OWN - NLM STAT- MEDLINE DCOM- 20160808 LR - 20220310 IS - 1528-0012 (Electronic) IS - 0016-5085 (Print) IS - 0016-5085 (Linking) VI - 150 IP - 4 DP - 2016 Apr TI - Lipid-Induced Signaling Causes Release of Inflammatory Extracellular Vesicles From Hepatocytes. PG - 956-67 LID - S0016-5085(15)01865-X [pii] LID - 10.1053/j.gastro.2015.12.037 [doi] AB - BACKGROUND & AIMS: Hepatocyte cellular dysfunction and death induced by lipids and macrophage-associated inflammation are characteristics of nonalcoholic steatohepatitis (NASH). The fatty acid palmitate can activate death receptor 5 (DR5) on hepatocytes, leading to their death, but little is known about how this process contributes to macrophage-associated inflammation. We investigated whether lipid-induced DR5 signaling results in the release of extracellular vesicles (EVs) from hepatocytes, and whether these can induce an inflammatory macrophage phenotype. METHODS: Primary mouse and human hepatocytes and Huh7 cells were incubated with palmitate, its metabolite lysophosphatidylcholine, or diluent (control). The released EV were isolated, characterized, quantified, and applied to macrophages. C57BL/6 mice were placed on chow or a diet high in fat, fructose, and cholesterol to induce NASH. Some mice also were given the ROCK1 inhibitor fasudil; 2 weeks later, serum EVs were isolated and characterized by immunoblot and nanoparticle-tracking analyses. Livers were collected and analyzed by histology, immunohistochemistry, and quantitative polymerase chain reaction. RESULTS: Incubation of primary hepatocytes and Huh7 cells with palmitate or lysophosphatidylcholine increased their release of EVs, compared with control cells. This release was reduced by inactivating mediators of the DR5 signaling pathway or rho-associated, coiled-coil-containing protein kinase 1 (ROCK1) inhibition. Hepatocyte-derived EVs contained tumor necrosis factor-related apoptosis-inducing ligand and induced expression of interleukin 1beta and interleukin 6 messenger RNAs in mouse bone marrow-derived macrophages. Activation of macrophages required DR5 and receptor-interacting protein kinase 1. Administration of the ROCK1 inhibitor fasudil to mice with NASH reduced serum levels of EVs; this reduction was associated with decreased liver injury, inflammation, and fibrosis. CONCLUSIONS: Lipids, which stimulate DR5, induce release of hepatocyte EVs, which activate an inflammatory phenotype in macrophages. Strategies to inhibit ROCK1-dependent release of EVs by hepatocytes might be developed for the treatment of patients with NASH. CI - Copyright (c) 2016 AGA Institute. Published by Elsevier Inc. All rights reserved. FAU - Hirsova, Petra AU - Hirsova P AD - Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester, Minnesota. FAU - Ibrahim, Samar H AU - Ibrahim SH AD - Division of Pediatric Gastroenterology and Hepatology, Mayo Clinic, Rochester, Minnesota. FAU - Krishnan, Anuradha AU - Krishnan A AD - Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester, Minnesota. FAU - Verma, Vikas K AU - Verma VK AD - Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester, Minnesota. FAU - Bronk, Steven F AU - Bronk SF AD - Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester, Minnesota. FAU - Werneburg, Nathan W AU - Werneburg NW AD - Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester, Minnesota. FAU - Charlton, Michael R AU - Charlton MR AD - Transplant Center, Intermountain Medical Center, Murray, Utah. FAU - Shah, Vijay H AU - Shah VH AD - Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester, Minnesota. FAU - Malhi, Harmeet AU - Malhi H AD - Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester, Minnesota. FAU - Gores, Gregory J AU - Gores GJ AD - Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester, Minnesota. Electronic address: gores.gregory@mayo.edu. LA - eng GR - R01 DK059615/DK/NIDDK NIH HHS/United States GR - U01 AA021788/AA/NIAAA NIH HHS/United States GR - KL2TR000136-09/TR/NCATS NIH HHS/United States GR - KL2 TR000136/TR/NCATS NIH HHS/United States GR - R01 DK041876/DK/NIDDK NIH HHS/United States GR - AA21788/AA/NIAAA NIH HHS/United States GR - DK41876/DK/NIDDK NIH HHS/United States GR - DK97178/DK/NIDDK NIH HHS/United States GR - P30DK084567/DK/NIDDK NIH HHS/United States GR - R37 DK041876/DK/NIDDK NIH HHS/United States GR - R37 AA021171/AA/NIAAA NIH HHS/United States GR - P30 DK084567/DK/NIDDK NIH HHS/United States GR - K08 DK097178/DK/NIDDK NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't DEP - 20160105 PL - United States TA - Gastroenterology JT - Gastroenterology JID - 0374630 RN - 0 (Inflammation Mediators) RN - 0 (Lysophosphatidylcholines) RN - 0 (Protein Kinase Inhibitors) RN - 0 (Receptors, TNF-Related Apoptosis-Inducing Ligand) RN - 0 (TNFRSF10B protein, human) RN - 0 (Tnfrsf10b protein, mouse) RN - 2V16EO95H1 (Palmitic Acid) RN - EC 2.7.11.1 (ROCK1 protein, human) RN - EC 2.7.11.1 (Rock1 protein, mouse) RN - EC 2.7.11.1 (rho-Associated Kinases) RN - EC 3.4.22.- (Caspases) SB - IM CIN - Gastroenterology. 2016 Apr;150(4):815-8. PMID: 26924096 MH - Animals MH - Caspases/metabolism MH - Cell Line, Tumor MH - Extracellular Vesicles/*drug effects/metabolism MH - HEK293 Cells MH - Hepatitis/drug therapy/*metabolism/pathology MH - Hepatocytes/*drug effects/metabolism/pathology MH - Humans MH - Inflammation Mediators/*metabolism MH - Liver/*drug effects/metabolism/pathology MH - Liver Cirrhosis, Experimental/metabolism/pathology MH - Lysophosphatidylcholines/*pharmacology MH - Macrophages/metabolism MH - Mice, Inbred C57BL MH - Mice, Transgenic MH - Non-alcoholic Fatty Liver Disease/drug therapy/*metabolism/pathology MH - Palmitic Acid/*pharmacology MH - Phenotype MH - Protein Kinase Inhibitors/pharmacology MH - RNA Interference MH - Rats, Sprague-Dawley MH - Receptors, TNF-Related Apoptosis-Inducing Ligand/agonists/genetics/metabolism MH - Signal Transduction/*drug effects MH - Transfection MH - rho-Associated Kinases/antagonists & inhibitors/genetics/metabolism PMC - PMC4808464 MID - NIHMS749108 OTO - NOTNLM OT - Cell Death OT - Exosomes OT - Lipotoxic OT - Microvesicles EDAT- 2016/01/15 06:00 MHDA- 2016/08/09 06:00 PMCR- 2017/04/01 CRDT- 2016/01/15 06:00 PHST- 2015/03/19 00:00 [received] PHST- 2015/12/24 00:00 [revised] PHST- 2015/12/29 00:00 [accepted] PHST- 2016/01/15 06:00 [entrez] PHST- 2016/01/15 06:00 [pubmed] PHST- 2016/08/09 06:00 [medline] PHST- 2017/04/01 00:00 [pmc-release] AID - S0016-5085(15)01865-X [pii] AID - 10.1053/j.gastro.2015.12.037 [doi] PST - ppublish SO - Gastroenterology. 2016 Apr;150(4):956-67. doi: 10.1053/j.gastro.2015.12.037. Epub 2016 Jan 5.