Kinaneh Safa, Hijaze Walaa, Mansour-Wattad Lana, Hammoud Rawan, Zaidani Hisam, Kabala Aviva, Hamoud Shadi
Department of Physiology, Rappaport Faculty of Medicine, Technion-Israel Institute of Technology, Haifa 3200003, Israel.
Department of Emergency Medicine, Rambam Health Care Campus, Haifa 3109601, Israel.
J Clin Med. 2022 Mar 17;11(6):1672. doi: 10.3390/jcm11061672.
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 E mice.
In vivo experiments: Male wild-type mice fed with either chow diet ( = 4) or high-fat diet ( = 6), and male E mice fed with chow diet ( = 8) or high-fat diet ( = 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, = 6) or high dose (13.3 mg/kg/week, ip, = 7), SST0001 (1.2 mg/mouse/day, ip, = 6), or normal saline (control, = 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 E mice aged 8-10 weeks and were cultured and treated with either PG545 or SST0001, both at 50 µg/mL for 24 h, followed by assessment of mRNA expression of steatosis related factors.
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.
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.
非酒精性脂肪性肝病影响着美国高达30%的成年人，并与慢性肝脏疾病的发病率和死亡率升高相关。肝脏脂肪变性的病理过程涉及多种分子途径，包括脂质摄取、脂肪生成、脂肪分解和β-氧化。乙酰肝素酶已被认为与肝脏脂肪变性有关。在此，我们研究了乙酰肝素酶抑制对E小鼠肝脏脂肪变性的影响。
体内实验：纳入雄性野生型小鼠，分别给予普通饮食（n = 4）或高脂饮食（n = 6），以及雄性E小鼠，分别给予普通饮食（n = 8）或高脂饮食（n = 33）。给予高脂饮食的小鼠用低剂量（6.4 mg/kg/周，腹腔注射，n = 6）或高剂量（13.3 mg/kg/周，腹腔注射，n = 7）的PG545、SST0001（1.2 mg/小鼠/天，腹腔注射，n = 6）或生理盐水（对照，n = 14）治疗12周。诱导腹膜炎两天后处死动物。分析血清中的生化参数。收集小鼠腹腔巨噬细胞（MPM）并分析其脂质含量。收获肝脏用于脂肪变性、脂质含量的组织病理学分析以及在mRNA水平分析脂肪变性相关因子的表达。体外实验：从8 - 10周龄未处理的E小鼠中分离MPM，用PG545或SST0001（均为50 μg/mL）培养和处理24小时，随后评估脂肪变性相关因子的mRNA表达。
乙酰肝素酶抑制显著减轻了肝脏脂肪变性的发展，这在肝脏组织学和脂质含量上很明显。血清分析表明，用乙酰肝素酶抑制剂处理的小鼠胆固醇和甘油三酯水平降低。在肝脏组织中，PG545处理后脂质摄取、脂肪分解、脂肪生成和β-氧化关键因子的mRNA表达评估显示显著下调，应用SST0001时下调程度较小。然而，用PG545而不是SST0001体外处理MPM导致这些细胞中的脂质含量增加，这与它们对处理小鼠的MPM的作用相反。这可能表明在乙酰肝素酶抑制后系统或分离的巨噬细胞中存在不同的调节途径。
乙酰肝素酶抑制通过降低组织脂质含量和影响关键脂质代谢调节因子的mRNA表达，显著减轻肝脏脂肪变性的发展。
