Voloshyna Tetyana, Holte Christopher, Pospíšil Jakub, Szafranska Karolina, Fuskevåg Ole Martin, Strand Sabina P, Åslund Andreas K O, Mørch Yrr, Snipstad Sofie, Sulheim Einar, Hunt Nicholas J, Cogger Victoria C, Couteur David G Le, Dietrichs Erik Sveberg, McCourt Peter A G
Vascular Biology Research Group, Dept. Medical Biology, University of Tromsø UiT The Arctic University of Norway, Tromsø, Norway.
Experimental and Clinical Pharmacology Research Group, Dept. Medical Biology, University of Tromsø UiT The Arctic University of Norway, Tromsø, Norway.
Nanotheranostics. 2025 May 14;9(2):155-170. doi: 10.7150/ntno.103000. eCollection 2025.
Ageing is established as the most significant risk factor for disease. About 75% of people over 75 years have diabetes or pre-diabetes and/or hyperlipidaemia which are established risk factors for cardiovascular outcomes, and risk factors for age-related conditions such as dementia, sarcopenia, frailty and osteoporosis. Age-related changes in the liver microcirculation, in particular relating to the cells lining the blood vessels, the liver sinusoidal endothelial cells (LSEC), are a potential cause for dyslipidaemia and insulin resistance in old age. There is also loss of LSEC mediated waste clearance functions essential for homeostasis. Finding ways to reverse these age-related changes in the LSEC will fill a significant gap in therapeutic options available for the treatment of ageing disorders. Such therapies may also benefit patients with fibrotic livers, since LSEC changes in this disease resemble those seen in the ageing LSEC in many aspects. Nanoparticles that access systemic circulation frequently accumulate in the liver. This could be utilized as a promising strategy for targeted drug delivery to the liver. The present study assessed if poly(alkyl-cyanoacrylate) nanoparticles (PACA NPs) are a suitable vector for the targeted transport of such therapeutics to LSEC, to reverse age-related changes such as fenestration/porosity loss. Mice were co-injected with PACA NPs and formaldehyde denatured serum albumin (FSA) and their livers were then examined by microscopy. PACA and FSA co-localised to LSEC, including at the sub-cellular level in endocytic vesicles. Isolated LSEC were challenged with Nile Red (NR668) labelled PACA NPs, which were rapidly internalized. HEK293 cells overexpressing stabilin-2 internalized PACA NPs, suggesting that stabilin-2 mediates PACA uptake on LSEC. Cultured LSEC from aged mice were challenged with PACA NPs containing sildenafil and examined with scanning electron microscopy to determine effects on fenestrations. Sildenafil PACA reversed age-related changes LSEC fenestration frequency and porosity at 3-fold lower sildenafil concentrations than sildenafil alone. If sildenafil PACA induces similar changes , age-related reduction of LSEC porosity could be reversed by the targeted delivery of sildenafil via PACA NPs.
衰老已被确认为疾病的最重要风险因素。75岁以上的人群中约75%患有糖尿病或糖尿病前期和/或高脂血症，这些都是心血管疾病的既定风险因素，也是与年龄相关疾病（如痴呆、肌肉减少症、虚弱和骨质疏松症）的风险因素。肝脏微循环中的年龄相关变化，特别是与血管内衬细胞（肝窦内皮细胞，LSEC）有关的变化，是老年血脂异常和胰岛素抵抗的潜在原因。LSEC介导的维持体内平衡所必需的废物清除功能也会丧失。找到逆转LSEC中这些与年龄相关变化的方法，将填补现有治疗衰老疾病的治疗选择中的重大空白。此类疗法可能也会使肝纤维化患者受益，因为这种疾病中LSEC的变化在许多方面与衰老LSEC中的变化相似。经常进入体循环的纳米颗粒会在肝脏中蓄积。这可作为一种有前景的策略用于肝脏的靶向药物递送。本研究评估了聚（烷基氰基丙烯酸酯）纳米颗粒（PACA NPs）是否是将此类治疗药物靶向转运至LSEC以逆转诸如窗孔/孔隙丧失等与年龄相关变化的合适载体。将小鼠同时注射PACA NPs和甲醛变性血清白蛋白（FSA），然后通过显微镜检查它们的肝脏。PACA和FSA共定位于LSEC，包括在内吞小泡的亚细胞水平。用尼罗红（NR668）标记的PACA NPs对分离的LSEC进行刺激，其被迅速内化。过表达稳定素-2的HEK293细胞内化PACA NPs，表明稳定素-2介导LSEC对PACA的摄取。用含有西地那非的PACA NPs对老年小鼠培养的LSEC进行刺激，并用扫描电子显微镜检查以确定对窗孔的影响。西地那非PACA在比单独使用西地那非低3倍的浓度下逆转了与年龄相关的LSEC窗孔频率和孔隙率变化。如果西地那非PACA诱导类似变化，那么通过PACA NPs靶向递送西地那非可以逆转与年龄相关的LSEC孔隙率降低。
