{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,11]],"date-time":"2026-02-11T15:38:37Z","timestamp":1770824317809,"version":"3.50.1"},"reference-count":55,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2022,3,17]],"date-time":"2022-03-17T00:00:00Z","timestamp":1647475200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["JCM"],"abstract":"<jats:p>Background: Non-alcoholic fatty liver disease affects up to 30% of adults in the USA, and is associated with a higher incidence of chronic liver morbidity and mortality. Several molecular pathways are involved in the pathology of liver steatosis, including lipid uptake, lipogenesis, lipolysis, and beta-oxidation. The enzyme heparanase has been implicated in liver steatosis. Herein, we investigated the effect of heparanase inhibition on liver steatosis in E0 mice. Methods: In vivo experiments: Male wild-type mice fed with either chow diet (n = 4) or high-fat diet (n = 6), and male E0 mice fed with chow diet (n = 8) or high-fat diet (n = 33) were included. Mice on a high-fat diet were treated for 12 weeks with PG545 at low dose (6.4 mg\/kg\/week, ip, n = 6) or high dose (13.3 mg\/kg\/week, ip, n = 7), SST0001 (1.2 mg\/mouse\/day, ip, n = 6), or normal saline (control, n = 14). Animals were sacrificed two days after inducing peritonitis. Serum was analyzed for biochemical parameters. Mouse peritoneal macrophages (MPMs) were harvested and analyzed for lipid content. Livers were harvested for histopathological analysis of steatosis, lipid content, and the expression of steatosis-related factors at the mRNA level. In vitro experiments: MPMs were isolated from untreated E0 mice aged 8\u201310 weeks and were cultured and treated with either PG545 or SST0001, both at 50 \u00b5g\/mL for 24 h, followed by assessment of mRNA expression of steatosis related factors. Results: Heparanase inhibition significantly attenuated the development of liver steatosis, as was evident by liver histology and lipid content. Serum analysis indicated lowering of cholesterol and triglycerides levels in mice treated with heparanase inhibitors. In liver tissue, assessment of mRNA expression of key factors in lipid uptake, lipolysis, lipogenesis, and beta-oxidation exhibited significant downregulation following PG545 treatment and to a lesser extent when SST0001 was applied. However, in vitro treatment of MPMs with PG545, but not SST0001, resulted in increased lipid content in these cells, which is opposed to their effect on MPMs of treated mice. This may indicate distinct regulatory pathways in the system or isolated macrophages following heparanase inhibition. Conclusion: Heparanase inhibition significantly attenuates the development of liver steatosis by decreasing tissue lipid content and by affecting the mRNA expression of key lipid metabolism regulators.<\/jats:p>","DOI":"10.3390\/jcm11061672","type":"journal-article","created":{"date-parts":[[2022,3,20]],"date-time":"2022-03-20T21:30:14Z","timestamp":1647811814000},"page":"1672","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Heparanase Inhibition Prevents Liver Steatosis in E0 Mice"],"prefix":"10.3390","volume":"11","author":[{"given":"Safa","family":"Kinaneh","sequence":"first","affiliation":[{"name":"Department of Physiology, Rappaport Faculty of Medicine, Technion\u2014Israel Institute of Technology, Haifa 3200003, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Walaa","family":"Hijaze","sequence":"additional","affiliation":[{"name":"Department of Emergency Medicine, Rambam Health Care Campus, Haifa 3109601, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lana","family":"Mansour-Wattad","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine E, Rambam Health Care Campus, Haifa 3109601, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rawan","family":"Hammoud","sequence":"additional","affiliation":[{"name":"Faculty of Biotechnology, Hadassah Academic College, Jerusalem 9101001, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hisam","family":"Zaidani","sequence":"additional","affiliation":[{"name":"Department of Emergency Medicine, Rambam Health Care Campus, Haifa 3109601, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Aviva","family":"Kabala","sequence":"additional","affiliation":[{"name":"Department of Physiology, Rappaport Faculty of Medicine, Technion\u2014Israel Institute of Technology, Haifa 3200003, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1405-239X","authenticated-orcid":false,"given":"Shadi","family":"Hamoud","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine E, Rambam Health Care Campus, Haifa 3109601, Israel"},{"name":"Lipid Research Laboratory, Rappaport Faculty of Medicine, Technion\u2014Israel Institute of Technology, Haifa 3200003, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,3,17]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"389","DOI":"10.1053\/j.gastro.2015.04.043","article-title":"Liver Fibrosis, but No Other Histologic Features, Is Associated with Long-term Outcomes of Patients with Nonalcoholic Fatty Liver Disease","volume":"149","author":"Angulo","year":"2015","journal-title":"Gastroenterology"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1547","DOI":"10.1002\/hep.27368","article-title":"Fibrosis stage is the strongest predictor for disease-specific mortality in NAFLD after up to 33 years of follow-up","volume":"61","author":"Ekstedt","year":"2015","journal-title":"Hepatology"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1002\/hep.28431","article-title":"Global epidemiology of nonalcoholic fatty liver disease- Meta-analytic assessment of prevalence, incidence, and outcomes","volume":"64","author":"Younossi","year":"2016","journal-title":"Hepatology"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1874","DOI":"10.1002\/hep.24268","article-title":"Pathologic criteria for nonalcoholic steatohepatitis: Interprotocol agreement and ability to predict liver-related mortality","volume":"53","author":"Younossi","year":"2011","journal-title":"Hepatology"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1186\/s13098-016-0169-x","article-title":"Empagliflozin (an SGLT2 inhibitor), alone or in combination with linagliptin (a DPP-4 inhibitor), prevents steatohepatitis in a novel mouse model of non-alcoholic steatohepatitis and diabetes","volume":"8","author":"Jojima","year":"2016","journal-title":"Diabetol. Metab. Syndr."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"785","DOI":"10.1002\/cphy.c120026","article-title":"Kupffer cells in the liver","volume":"3","author":"Dixon","year":"2013","journal-title":"Compr. Physiol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1473","DOI":"10.1002\/cphy.c120035","article-title":"Hepatic stellate cells and liver fibrosis","volume":"3","author":"Puche","year":"2013","journal-title":"Compr. Physiol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"e1920294","DOI":"10.1001\/jamanetworkopen.2019.20294","article-title":"Trends in Liver Disease Etiology Among Adults Awaiting Liver Transplantation in the United States, 2014\u20132019","volume":"3","author":"Wong","year":"2020","journal-title":"JAMA Netw. Open"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"697","DOI":"10.3945\/an.113.004648","article-title":"Hepatic fatty acid trafficking: Multiple forks in the road","volume":"4","author":"Mashek","year":"2013","journal-title":"Adv. Nutr."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2238","DOI":"10.1016\/j.ajpath.2016.06.003","article-title":"Pathogenesis of Kupffer Cells in Cholestatic Liver Injury","volume":"186","author":"Sato","year":"2016","journal-title":"Am. J. Pathol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"8323747","DOI":"10.1155\/2016\/8323747","article-title":"Regulation of Hepatic Stellate Cells and Fibrogenesis by Fibroblast Growth Factors","volume":"2016","author":"Schumacher","year":"2016","journal-title":"BioMed Res. Int."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1653","DOI":"10.1016\/S0002-9440(10)63522-5","article-title":"Role of fibroblast growth factor type 1 and 2 in carbon tetrachloride-induced hepatic injury and fibrogenesis","volume":"163","author":"Yu","year":"2003","journal-title":"Am. J. Pathol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"181","DOI":"10.1038\/nri3623","article-title":"Liver fibrosis and repair: Immune regulation of wound healing in a solid organ","volume":"14","author":"Pellicoro","year":"2014","journal-title":"Nat. Rev. Immunol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"547","DOI":"10.1053\/j.gastro.2014.11.039","article-title":"Nonalcoholic Steatohepatitis Is the Second Leading Etiology of Liver Disease Among Adults Awaiting Liver Transplantation in the United States","volume":"148","author":"Wong","year":"2015","journal-title":"Gastroenterology"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1249","DOI":"10.1053\/j.gastro.2011.06.061","article-title":"Frequency and outcomes of liver transplantation for nonalcoholic steatohepatitis in the United States","volume":"141","author":"Charlton","year":"2011","journal-title":"Gastroenterology"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1314","DOI":"10.1038\/ajg.2013.160","article-title":"Population-attributable fractions of risk factors for hepatocellular carcinoma in the United States","volume":"108","author":"Welzel","year":"2013","journal-title":"Am. J. Gastroenterol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1353","DOI":"10.1016\/j.jhep.2011.07.010","article-title":"A role for autophagy during hepatic stellate cell activation","volume":"55","author":"Thoen","year":"2011","journal-title":"J. Hepatol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1405","DOI":"10.3892\/ijmm.2017.3145","article-title":"Autophagy mediated by endoplasmic reticulum stress enhances the caffeine-induced apoptosis of hepatic stellate cells","volume":"40","author":"Li","year":"2017","journal-title":"Int. J. Mol. Med."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2063","DOI":"10.1056\/NEJMra1503519","article-title":"Cause, Pathogenesis, and Treatment of Nonalcoholic Steatohepatitis","volume":"377","author":"Diehl","year":"2017","journal-title":"N. Engl. J. Med."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1294","DOI":"10.1007\/s10620-016-4049-x","article-title":"Innate Immunity and Inflammation in NAFLD\/NASH","volume":"61","author":"Arrese","year":"2016","journal-title":"Dig. Dis. Sci."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"88","DOI":"10.1038\/nrgastro.2015.200","article-title":"Immunology in the liver\u2014From homeostasis to disease","volume":"13","author":"Heymann","year":"2016","journal-title":"Nat. Rev. Gastroenterol. Hepatol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"138","DOI":"10.1016\/j.hepres.2005.09.022","article-title":"Lessons from animal models of NASH","volume":"33","author":"Diehl","year":"2005","journal-title":"Hepatol. Res. Off. J. Jpn. Soc. Hepatol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"229","DOI":"10.1002\/hep.24368","article-title":"Liver-Specific Loss of Glucose-Regulated Protein 78 Perturbs the Unfolded Protein Response and Exacerbates a Spectrum of Liver Diseases in Mice","volume":"54","author":"Ji","year":"2011","journal-title":"Hepatology"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"427","DOI":"10.1111\/j.1582-4934.2007.00039.x","article-title":"Mammalian heparanase: What is the message?","volume":"11","author":"Vreys","year":"2007","journal-title":"J. Cell. Mol. Med."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"415","DOI":"10.1515\/bmc-2015-0021","article-title":"Recent data concerning heparanase: Focus on fibrosis, inflammation and cancer","volume":"6","author":"Secchi","year":"2015","journal-title":"Biomol. Concepts"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"2882","DOI":"10.1097\/01.ASN.0000142426.55612.6D","article-title":"A synthetic heparanase inhibitor reduces proteinuria in passive Heymann nephritis","volume":"15","author":"Levidiotis","year":"2004","journal-title":"J. Am. Soc. Nephrol. JASN"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1038\/nrneph.2017.6","article-title":"Heparanase: Roles in cell survival, extracellular matrix remodelling and the development of kidney disease","volume":"13","author":"Rabelink","year":"2017","journal-title":"Nat. Rev. Nephrol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"S90","DOI":"10.1016\/S0049-3848(14)50015-1","article-title":"Heparanase multiple effects in cancer","volume":"133","author":"Nadir","year":"2014","journal-title":"Thromb. Res."},{"key":"ref_29","first-page":"54","article-title":"Heparanase: From basic research to therapeutic applications in cancer and inflammation","volume":"29","author":"Vlodavsky","year":"2016","journal-title":"Drug Resist. Updates Rev. Comment. Antimicrob. Anticancer. Chemother."},{"key":"ref_30","first-page":"2122","article-title":"Heparanase is a key player in renal fibrosis by regulating TGF-beta expression and activity","volume":"1843","author":"Masola","year":"2014","journal-title":"Bba-Mol. Cell Res."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1478","DOI":"10.1074\/jbc.M111.279836","article-title":"Heparanase and syndecan-1 interplay orchestrates fibroblast growth factor-2-induced epithelial-mesenchymal transition in renal tubular cells","volume":"287","author":"Masola","year":"2012","journal-title":"J. Biol. Chem."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"208","DOI":"10.2337\/db11-1024","article-title":"Heparanase Is Essential for the Development of Diabetic Nephropathy in Mice","volume":"61","author":"Gil","year":"2012","journal-title":"Diabetes"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"11384","DOI":"10.3748\/wjg.v20.i32.11384","article-title":"Adjuvant heparanase inhibitor PI-88 therapy for hepatocellular carcinoma recurrence","volume":"20","author":"Liu","year":"2014","journal-title":"World J. Gastroenterol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"192","DOI":"10.1016\/S1386-6346(03)00107-4","article-title":"Heparanase expression in hepatocellular carcinoma and the cirrhotic liver","volume":"26","author":"Xiao","year":"2003","journal-title":"Hepatol. Res. Off. J. Jpn. Soc. Hepatol."},{"key":"ref_35","first-page":"1299","article-title":"The clinicopathological significance of heparanase and basic fibroblast growth factor expressions in hepatocellular carcinoma","volume":"7","author":"Yamanoi","year":"2001","journal-title":"Clin. Cancer Res. Off. J. Am. Assoc. Cancer Res."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1016\/S0959-8049(02)00558-0","article-title":"Heparanase gene expression and its correlation with spontaneous apoptosis in hepatocytes of cirrhotic liver and carcinoma","volume":"39","author":"Ikeguchi","year":"2003","journal-title":"Eur. J. Cancer"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"7357495","DOI":"10.1155\/2017\/7357495","article-title":"Heparanase Inhibition Reduces Glucose Levels, Blood Pressure, and Oxidative Stress in Apolipoprotein E Knockout Mice","volume":"2017","author":"Hamoud","year":"2017","journal-title":"BioMed. Res. Int."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Shekh Muhammad, R., Niroz, A.S., Kinaneh, S., Agbaria, M., Sabo, E., Grajeda-Iglesias, C., Volkova, N., and Hamoud, S. (2018). Heparanase inhibition attenuates atherosclerosis progression and liver steatosis in E0 mice. Atherosclerosis, in press.","DOI":"10.1016\/j.atherosclerosis.2018.07.026"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1382","DOI":"10.1158\/1078-0432.CCR-10-2476","article-title":"SST0001, a chemically modified heparin, inhibits myeloma growth and angiogenesis via disruption of the heparanase\/syndecan-1 axis","volume":"17","author":"Ritchie","year":"2011","journal-title":"Clin. Cancer Res. Off. J. Am. Assoc. Cancer Res."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1857","DOI":"10.1111\/j.1582-4934.2010.01190.x","article-title":"Pre-clinical and clinical significance of heparanase in Ewing\u2019s sarcoma","volume":"15","author":"Shafat","year":"2011","journal-title":"J. Cell Mol. Med."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1424","DOI":"10.1016\/j.bcp.2013.02.023","article-title":"Antitumor efficacy of the heparanase inhibitor SST0001 alone and in combination with antiangiogenic agents in the treatment of human pediatric sarcoma models","volume":"85","author":"Cassinelli","year":"2013","journal-title":"Biochem. Pharmacol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"302","DOI":"10.1016\/S0076-6879(78)52032-6","article-title":"Microsomal lipid peroxidation","volume":"52","author":"Buege","year":"1978","journal-title":"Methods Enzym."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"244","DOI":"10.1016\/S0076-6879(01)35247-3","article-title":"Markers for low-density lipoprotein oxidation","volume":"335","author":"Aviram","year":"2001","journal-title":"Methods Enzym."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1016\/S0021-9258(19)52451-6","article-title":"Protein measurement with the Folin phenol reagent","volume":"193","author":"Lowry","year":"1951","journal-title":"J. Biol. Chem."},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Planer, D., Metzger, S., Zcharia, E., Wexler, I.D., Vlodavsky, I., and Chajek-Shaul, T. (2011). Role of Heparanase on Hepatic Uptake of Intestinal Derived Lipoprotein and Fatty Streak Formation in Mice. PLoS ONE, 6.","DOI":"10.1371\/journal.pone.0018370"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1172\/JCI29154","article-title":"Liver heparan sulfate proteoglycans mediate clearance of triglyceride-rich lipoproteins independently of LDL receptor family members","volume":"117","author":"MacArthur","year":"2007","journal-title":"J. Clin. Investig."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"396","DOI":"10.1097\/MOL.0b013e328338cabc","article-title":"Recent insights into factors affecting remnant lipoprotein uptake","volume":"21","author":"Williams","year":"2010","journal-title":"Curr. Opin. Lipidol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1097\/00041433-199710000-00003","article-title":"Cell-surface heparan sulfate proteoglycans: Dynamic molecules mediating ligand catabolism","volume":"8","author":"Williams","year":"1997","journal-title":"Curr. Opin. Lipidol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1007\/s00394-013-0516-8","article-title":"Epigallocatechin gallate attenuates fibrosis, oxidative stress, and inflammation in non-alcoholic fatty liver disease rat model through TGF\/SMAD, PI3 K\/Akt\/FoxO1, and NF-kappa B pathways","volume":"53","author":"Xiao","year":"2014","journal-title":"Eur. J. Nutr."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1016\/j.placenta.2018.03.008","article-title":"Cadmium induces inflammatory cytokines through activating Akt signaling in mouse placenta and human trophoblast cells","volume":"65","author":"Hu","year":"2018","journal-title":"Placenta"},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"Zhao, Y., Qin, W., Zhang, J.P., Hu, Z.Y., Tong, J.W., Ding, C.B., Peng, Z.G., Zhao, L.X., Song, D.Q., and Jiang, J.D. (2013). HCV IRES-mediated core expression in zebrafish. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0056985"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"14956","DOI":"10.1038\/s41598-017-14946-0","article-title":"Heparanase and macrophage interplay in the onset of liver fibrosis","volume":"7","author":"Secchi","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"530","DOI":"10.1111\/j.1365-2249.1994.tb07031.x","article-title":"IL-6 induces hepatic inflammation and collagen synthesis in vivo","volume":"95","author":"Choi","year":"1994","journal-title":"Clin. Exp. Immunol."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"4234061","DOI":"10.1155\/2016\/4234061","article-title":"Insights into the Role and Interdependence of Oxidative Stress and Inflammation in Liver Diseases","volume":"2016","author":"Li","year":"2016","journal-title":"Oxid Med. Cell Longev."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"26087","DOI":"10.3390\/ijms161125942","article-title":"The Role of Oxidative Stress and Antioxidants in Liver Diseases","volume":"16","author":"Li","year":"2015","journal-title":"Int. J. Mol. Sci."}],"container-title":["Journal of Clinical Medicine"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2077-0383\/11\/6\/1672\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:38:10Z","timestamp":1760135890000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2077-0383\/11\/6\/1672"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,3,17]]},"references-count":55,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2022,3]]}},"alternative-id":["jcm11061672"],"URL":"https:\/\/doi.org\/10.3390\/jcm11061672","relation":{},"ISSN":["2077-0383"],"issn-type":[{"value":"2077-0383","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,3,17]]}}}