载阿立新-13 的巨噬细胞膜包裹纳米粒通过抑制 NLRP3 炎性小体介导的细胞焦亡靶向治疗缺血性脑卒中。
Apelin-13-Loaded Macrophage Membrane-Encapsulated Nanoparticles for Targeted Ischemic Stroke Therapy via Inhibiting NLRP3 Inflammasome-Mediated Pyroptosis.
机构信息
Department of Basic Medical Sciences, Shandong Second Medical University, Weifang, Shandong, People's Republic of China.
Department of Ophthalmology, Affiliated Hospital of Shandong Second Medical University, Weifang, People's Republic of China.
出版信息
Int J Nanomedicine. 2024 Sep 7;19:9175-9193. doi: 10.2147/IJN.S475915. eCollection 2024.
PURPOSE
Ischemic stroke is a refractory disease wherein the reperfusion injury caused by sudden restoration of blood supply is the main cause of increased mortality and disability. However, current therapeutic strategies for the inflammatory response induced by cerebral ischemia-reperfusion (I/R) injury are unsatisfactory. This study aimed to develop a functional nanoparticle (MM/ANPs) comprising apelin-13 (APNs) encapsulated in macrophage membranes (MM) modified with distearoyl phosphatidylethanolamine-polyethylene glycol-RVG29 (DSPE-PEG-RVG29) to achieve targeted therapy against ischemic stroke.
METHODS
MM were extracted from RAW264.7. PLGA was dissolved in dichloromethane, while Apelin-13 was dissolved in water, and CY5.5 was dissolved in dichloromethane. The precipitate was washed twice with ultrapure water and then resuspended in 10 mL to obtain an aqueous solution of PLGA nanoparticles. Subsequently, the cell membrane was evenly dispersed homogeneously and mixed with PLGA-COOH at a mass ratio of 1:1 for the hybrid ultrasound. DSPE-PEG-RVG29 was added and incubated for 1 h to obtain MM/ANPs.
RESULTS
In this study, we developed a functional nanoparticle delivery system (MM/ANPs) that utilizes macrophage membranes coated with DSPE-PEG-RVG29 peptide to efficiently deliver Apelin-13 to inflammatory areas using ischemic stroke therapy. MM/ANPs effectively cross the blood-brain barrier and selectively accumulate in ischemic and inflamed areas. In a mouse I/R injury model, these nanoparticles significantly improved neurological scores and reduced infarct volume. Apelin-13 is gradually released from the MM/ANPs, inhibiting NLRP3 inflammasome assembly by enhancing sirtuin 3 (SIRT3) activity, which suppresses the inflammatory response and pyroptosis. The positive regulation of SIRT3 further inhibits the NLRP3-mediated inflammation, showing the clinical potential of these nanoparticles for ischemic stroke treatment. The biocompatibility and safety of MM/ANPs were confirmed through in vitro cytotoxicity tests, blood-brain barrier permeability tests, biosafety evaluations, and blood compatibility studies.
CONCLUSION
MM/ANPs offer a highly promising approach to achieve ischemic stroke-targeted therapy inhibiting NLRP3 inflammasome-mediated pyroptosis.
目的
缺血性脑卒中是一种难治性疾病，突然恢复血液供应引起的再灌注损伤是导致死亡率和残疾率增加的主要原因。然而，目前针对脑缺血再灌注（I/R）损伤引起的炎症反应的治疗策略并不令人满意。本研究旨在开发一种功能性纳米颗粒（MM/ANPs），该纳米颗粒由包裹在巨噬细胞膜（MM）中的阿皮素-13（APNs）组成，MM 经过二硬脂酰基磷脂酰乙醇胺-聚乙二醇-RVG29（DSPE-PEG-RVG29）修饰，以实现针对缺血性脑卒中的靶向治疗。
方法
从 RAW264.7 中提取 MM。PLGA 溶解在二氯甲烷中，阿皮素-13 溶解在水中，CY5.5 溶解在二氯甲烷中。将沉淀物用超纯水洗涤两次，然后重悬于 10 mL 水中，得到 PLGA 纳米颗粒的水溶液。然后，将细胞膜均匀分散并与 PLGA-COOH 以 1:1 的质量比混合进行杂交超声。加入 DSPE-PEG-RVG29 并孵育 1 小时，得到 MM/ANPs。
结果
在本研究中，我们开发了一种功能性纳米颗粒递药系统（MM/ANPs），该系统利用涂有 DSPE-PEG-RVG29 肽的巨噬细胞膜，通过缺血性脑卒中治疗，将阿皮素-13 高效递送至炎症区域。MM/ANPs 可有效穿透血脑屏障，并选择性地在缺血和炎症区域积聚。在小鼠 I/R 损伤模型中，这些纳米颗粒显著改善了神经功能评分并减少了梗死体积。阿皮素-13 从 MM/ANPs 中逐渐释放，通过增强 SIRT3 活性抑制 NLRP3 炎性小体的组装，从而抑制炎症反应和细胞焦亡。SIRT3 的正调控进一步抑制了 NLRP3 介导的炎症反应，显示了这些纳米颗粒在缺血性脑卒中治疗中的临床潜力。通过体外细胞毒性试验、血脑屏障通透性试验、生物安全性评价和血液相容性研究证实了 MM/ANPs 的生物相容性和安全性。
结论
MM/ANPs 为实现缺血性脑卒中靶向治疗、抑制 NLRP3 炎性小体介导的细胞焦亡提供了一种很有前途的方法。
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