天麻素修饰的树枝状大分子包裹金纳米粒子作为脑缺血再灌注损伤的药物传递系统的制备。

Preparation of gastrodin-modified dendrimer-entrapped gold nanoparticles as a drug delivery system for cerebral ischemia-reperfusion injury.

机构信息

Department of Neurosurgery, First Affiliated Hospital of Kunming Medical University, Kunming, China.

出版信息

Brain Behav. 2022 Dec;12(12):e2810. doi: 10.1002/brb3.2810. Epub 2022 Nov 21.

DOI:10.1002/brb3.2810

PMID:36408880

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9759136/

Abstract

OBJECTIVE

This study sought to evaluate the feasibility of multifunctional gastrodin (GAS)-containing nano-drug carrier system against cerebral ischemia-reperfusion injury (CIRI).

METHODS

The drug-loaded nanocomposite (Au-G5.NHAc-PS/GAS) with certain encapsulation efficiency (EE) was prepared by physical adsorption method using different proportions of GAS and drug-carrying system (Au-G5.NHAc-PS). High-performance liquid chromatography was used to determine the drug loading and EE. Cultured rat astrocytes and hypothalamic neurons were assigned into four groups: PBS, Au-G5.NHAc-PS, Au-G5.NHAc-PS/GAS, and GAS. CCK-8 assay, flow cytometry, and quantitative real-time PCR were performed to examine the cell viability, apoptosis, and the expression of tumor necrosis factor-α (TNF-α), IL-1β, and IL-6 in the astrocytes and hypothalamic neurons, respectively. Cellular uptake of GAS and Au-G5.NHAc-PS/GAS was analyzed by using Hoechst 33342 staining. The animal model with focal cerebral ischemia was generated by middle cerebral artery occlusion (MCAO) in healthy male Sprague Dawley (SD) rats, and pathological changes of brain tissue and major organs in the rats were identified by hematoxylin and eosin (HE) staining. Apoptosis in rat astrocytes and hypothalamic neurons was detected by TUNEL staining and flow cytometry.

RESULTS

Au-G5.NHAc-PS had a spherical shape with a uniform size of 157.3 nm. Among the nanoparticles, Au-G5.NHAc-PS/GAS with an EE of 70.3% displayed the best release delay effect. Moreover, we observed that in vitro cytotoxicity and cellular uptake of Au-G5.NHAc-PS/GAS were higher than those of GAS, whereas the expression of TNF-α, IL-1β, and IL-6 was significantly downregulated in Au-G5.NHAc-PS/GAS group as compared to G5.NHAc-PS group. Notably, HE staining revealed that although Au-G5.NHAc-PS/GAS had no toxic and side effects on the main organs of rats, it alleviated the damage of brain tissue in the MCAO rats. Besides, Au-G5.NHAc/GAS markedly reduced MCAO-induced apoptosis.

CONCLUSION

Au-G5.NHAc-PS showed favorable surface morphology, sustained drug release ability, no measurable toxicity, and good biocompatibility, indicating that GAS exerts anti-inflammatory and antiapoptotic effects on CIRI.

摘要

目的

本研究旨在评估载天麻素（GAS）的多功能纳米药物载体系统对脑缺血再灌注损伤（CIRI）的可行性。

方法

采用物理吸附法，用不同比例的 GAS 和载药系统（Au-G5.NHAc-PS）制备载药纳米复合物（Au-G5.NHAc-PS/GAS），并具有一定的包封率（EE）。采用高效液相色谱法测定药物的载药量和 EE。将培养的大鼠星形胶质细胞和下丘脑神经元分为四组：PBS、Au-G5.NHAc-PS、Au-G5.NHAc-PS/GAS 和 GAS。CCK-8 测定法、流式细胞术和实时定量 PCR 分别检测星形胶质细胞和下丘脑神经元的细胞活力、细胞凋亡以及肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）和白细胞介素-6（IL-6）的表达。采用 Hoechst 33342 染色分析 GAS 和 Au-G5.NHAc-PS/GAS 的细胞摄取情况。通过大脑中动脉闭塞（MCAO）建立健康雄性 Sprague Dawley（SD）大鼠局灶性脑缺血动物模型，采用苏木精和伊红（HE）染色观察大鼠脑组织和主要器官的病理变化。TUNEL 染色和流式细胞术检测大鼠星形胶质细胞和下丘脑神经元的细胞凋亡。

结果

Au-G5.NHAc-PS 呈球形，粒径均匀，大小为 157.3nm。在纳米粒子中，EE 为 70.3%的 Au-G5.NHAc-PS/GAS 显示出最佳的释放延迟效果。此外，我们观察到 Au-G5.NHAc-PS/GAS 的体外细胞毒性和细胞摄取均高于 GAS，而 Au-G5.NHAc-PS/GAS 组的 TNF-α、IL-1β 和 IL-6 表达明显低于 G5.NHAc-PS 组。值得注意的是，HE 染色显示，尽管 Au-G5.NHAc-PS/GAS 对大鼠主要器官没有毒性和副作用，但它减轻了 MCAO 大鼠脑组织的损伤。此外，Au-G5.NHAc-PS/GAS 显著减少了 MCAO 诱导的细胞凋亡。