Shiraz Institute for Stem Cell & Regenerative Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.
Department of Pharmaceutical Nanotechnology, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran.
J Cell Mol Med. 2024 Jul;28(13):e18529. doi: 10.1111/jcmm.18529.
In this in vitro study, for the first time, we evaluate the effects of simvastatin-loaded liposome nanoparticles (SIM-LipoNPs) treatment on fibrosis-induced liver microtissues, as simvastatin (SIM) has shown potential benefits in the non-alcoholic fatty liver disease process. We developed multicellular liver microtissues composed of hepatic stellate cells, hepatoblastoma cells and human umbilical vein endothelial cells. The microtissues were supplemented with a combination of palmitic acid and oleic acid to develop fibrosis models. Subsequently, various groups of microtissues were exposed to SIM and SIM-LipoNPs at doses of 5 and 10 mg/mL. The effectiveness of the treatments was evaluated by analysing cell viability, production of reactive oxygen species (ROS) and nitric oxide (NO), the expression of Kruppel-like factor (KLF) 2, and pro-inflammatory cytokines (interleukin(IL)-1 α, IL-1 β, IL-6 and tumour necrosis factor-α), and the expression of collagen I. Our results indicated that SIM-LipoNPs application showed promising results. SIM-LipoNPs effectively amplified the SIM-klf2-NO pathway at a lower dosage compatible with a high dosage of free SIM, which also led to reduced oxidative stress by decreasing ROS levels. SIM-LipoNPs administration also resulted in a significant reduction in pro-inflammatory cytokines and Collagen I mRNA levels, as a marker of fibrosis. In conclusion, our study highlights the considerable therapeutic potential of using SIM-LipoNPs to prevent liver fibrosis progress, underscoring the remarkable properties of SIM-LipoNPs in activating the KLF2-NO pathway and anti-oxidative and anti-inflammatory response.
在这项体外研究中，我们首次评估了辛伐他汀负载脂质体纳米颗粒（SIM-LipoNPs）治疗对纤维化诱导的肝组织的影响，因为辛伐他汀（SIM）在非酒精性脂肪性肝病过程中显示出了潜在的益处。我们开发了由肝星状细胞、肝癌细胞和人脐静脉内皮细胞组成的多细胞肝组织。这些组织补充了棕榈酸和油酸的组合，以建立纤维化模型。随后，将不同组的微组织暴露于 5 和 10mg/mL 的 SIM 和 SIM-LipoNPs 中。通过分析细胞活力、活性氧（ROS）和一氧化氮（NO）的产生、Kruppel 样因子（KLF）2 的表达以及促炎细胞因子（白细胞介素（IL）-1α、IL-1β、IL-6 和肿瘤坏死因子-α）和胶原 I 的表达来评估治疗效果。我们的结果表明，SIM-LipoNPs 的应用显示出了有希望的结果。SIM-LipoNPs 以与游离 SIM 高剂量相容的低剂量有效地放大了 SIM-KLF2-NO 通路，这也通过降低 ROS 水平来减少氧化应激。SIM-LipoNPs 给药还导致促炎细胞因子和胶原 I mRNA 水平的显著降低，作为纤维化的标志物。总之，我们的研究强调了使用 SIM-LipoNPs 预防肝纤维化进展的巨大治疗潜力，突出了 SIM-LipoNPs 在激活 KLF2-NO 通路和抗氧化及抗炎反应方面的显著特性。
