Department of Biochemistry, Sector-25, Panjab University, Chandigarh 160014, India.
CSIR, Central Scientific Instruments Organization, Sector-30-C, Chandigarh 160030, India.
Chem Phys Lipids. 2022 Aug;246:105207. doi: 10.1016/j.chemphyslip.2022.105207. Epub 2022 May 24.
Hepcidin, a key regulator of iron homeostasis, has been implicated in the pathogenesis of various iron-related diseases. Although small interfering RNA (siRNA) are potent to modulate the expression of hepcidin, their bioavailability remains a major issue. The β-galactopyranoside-conjugated liposomes (GAL-liposome) targeting liver synthesized hepcidin were prepared by thin lipid film hydration method to encapsulate siRNA and the conjugation of β-galactopyranoside to the lipid nanocarrier was achieved by covalent chemistry. The prepared siRNA loaded GAL-lip were spherical with around 50 nm radius in size as observed by HR-TEM. The zeta potential and polydispersity index of the prepared liposomes were - 19.9 ± 0.96 mV and 0.44 ± 0.05, respectively. The encapsulation efficiency as determined by dialysis bag method was around 91.76 ± 1.74%. The cell viability and cellular uptake analysis was examined in HepG2 cells by MTT assay and flow cytometry, respectively. The stability and cumulative release of siRNA was also assessed. The hepcidin mRNA expression on administration of siRNA loaded GAL-lip was determined in HepG2 cells and in lipopolysaccharide-induced mice model followed by examining itsin vivo biodistribution by fluorescence microscopy. The results suggested thatsiRNA loaded GAL-lip reduced the hepcidin levels, thus, highlighting a novel ligand conjugated ionizable lipid-based nanocarrier for inducing RNA interference.
亚铁调素是铁稳态的关键调节剂，与各种铁相关疾病的发病机制有关。虽然小干扰 RNA (siRNA) 能够有效地调节亚铁调素的表达，但它们的生物利用度仍然是一个主要问题。通过薄脂质膜水化法制备了靶向肝脏合成亚铁调素的β-半乳糖吡喃糖苷缀合脂质体（GAL-脂质体），以包封 siRNA，并通过共价化学将β-半乳糖吡喃糖苷缀合到脂质纳米载体上。通过高分辨率透射电子显微镜观察到，制备的负载 siRNA 的 GAL-脂质体呈球形，粒径约为 50nm。制备的脂质体的 zeta 电位和多分散指数分别为-19.9±0.96mV 和 0.44±0.05。通过透析袋法测定的包封效率约为 91.76±1.74%。通过 MTT 测定法和流式细胞术分别在 HepG2 细胞中进行细胞活力和细胞摄取分析。还评估了 siRNA 的稳定性和累积释放。通过荧光显微镜检查，在 HepG2 细胞和脂多糖诱导的小鼠模型中测定了给药后负载 siRNA 的 GAL-脂质体上的亚铁调素 mRNA 表达，并检查了其体内分布。结果表明，负载 siRNA 的 GAL-脂质体降低了亚铁调素水平，从而突出了一种新型配体偶联可离子化脂质基纳米载体，用于诱导 RNA 干扰。
