半影靶向CircOGDH siRNA载体纳米粒子可减轻局灶性脑缺血的神经细胞凋亡。

IF 4.4 1区 医学 Q1 CLINICAL NEUROLOGY
Yanfang Liu, Tianyuan Zhang, Xing Zou, Zhongwen Yuan, Yufeng Li, Jiankun Zang, Niu He, Lizhen He, Anding Xu, Dan Lu
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引用次数: 0

摘要

背景:纳米颗粒(NPs)是一类可装载治疗药物并输送到特定区域的物质。在早前的研究中,我们发现神经元衍生的环状 RNA(circRNA)--环状氧戊二酸脱氢酶(CircOGDH)--是治疗急性缺血性中风的一个很有前景的靶点。本研究致力于探索将基于 CircOGDH 的 NP 运送到大脑中动脉闭塞/再灌注(MCAO/R)小鼠缺血半影区的前瞻性初步策略:方法:原代皮层神经元免疫荧光和体内荧光成像显示了聚乳酸共聚乙二醇(PLGA)聚酰胺胺(PAMAM)@CircOGDH小干扰RNA(siRNA)NPs的内吞作用。Western 印迹分析和 CCK8 检测评估了用 PLGA-PAMAM@CircOGDH siRNA NPs 处理的缺血性神经元的凋亡水平。通过定量反转录 PCR 实验、小鼠行为测试、T2 MRI 分析、Nissl 和 TdT 介导的 dUTP 缺口标记(TUNEL)联合染色来评估 MCAO/R 小鼠缺血半影神经元的凋亡水平。通过血常规检查、肝肾功能检查和 HE 染色检测 NPs 对 MCAO/R 小鼠的生物安全性:结果:PLGA-PAMAM@CircOGDH siRNA NPs组装成功。结果:PLGA-PAMAM@CircOGDH siRNA NPs成功组装,其在缺血神经元中的内吞作用减轻了神经元在体外和体内的凋亡水平。此外,小鼠行为测试表明,尾部注射 PLGA-PAMAM@CircOGDH siRNA NPs 后,MCAO/R 小鼠的神经系统缺陷明显缓解,且未观察到毒性作用:总之,我们的研究结果表明,PLGA-PAMAM@CircOGDH siRNA NPs可被递送至缺血半影区,缓解MCAO/R小鼠和缺血性神经元的凋亡;因此,我们的研究为使用基于circRNA的NPs治疗缺血性脑卒中提供了一种理想的方法。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Penumbra-targeted CircOGDH siRNA-loaded nanoparticles alleviate neuronal apoptosis in focal brain ischaemia.

Background: Nanoparticles (NPs) are a class of substances that can be loaded with therapeutic agents delivered to specific areas. In our earlier research, we identified a neuron-derived circular RNA (circRNA), circular oxoglutarate dehydrogenase (CircOGDH), as a promising therapeutic target for acute ischaemic stroke. This study dedicated to explore a prospective preliminary strategy of CircOGDH-based NP delivered to the ischaemic penumbra region in middle cerebral artery occlusion/reperfusion (MCAO/R) mice.

Methods: Immunofluorescence in primary cortex neurons and in vivo fluorescence imaging revealed endocytosis of Poly(lactide-co-glycolide) (PLGA) poly amidoamine(PAMAM)@CircOGDH small interfering RNA (siRNA) NPs. Western blotting analysis and CCK8 assay were performed to evaluate the apoptotic level in ischaemic neurons treated with PLGA-PAMAM@CircOGDH siRNA NPs. Quantitative reverse transcription PCR experiments, mice behaviour test, T2 MRI analysis, Nissl and TdT-mediated dUTP nick end labeling (TUNEL) co-staining were performed to evaluate the apoptosis level of ischaemic penumbra neurons in MCAO/R mice. Biosafety evaluation of NPs in MCAO/R mice was detected by blood routine examination, liver and kidney function examination and HE staining.

Results: PLGA-PAMAM@CircOGDH siRNA NPs were successfully assembled. Endocytosis of PLGA-PAMAM@CircOGDH siRNA NPs in ischaemic neurons alleviated neuronal apoptotic level in vitro and in vivo. Furthermore, mice behaviour test showed that the neurological defects of MCAO/R mice were significantly alleviated after the tail injection of PLGA-PAMAM@CircOGDH siRNA NPs, and no toxic effects were observed.

Conclusion: In conclusion, our results suggest that PLGA-PAMAM@CircOGDH siRNA NPs can be delivered to the ischaemic penumbra region and alleviate neuron apoptosis in MCAO/R mice and in ischaemic neurons; therefore, our study provides a desirable approach for using circRNA-based NPs for the treatment of ischaemic stroke.

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来源期刊
Stroke and Vascular Neurology
Stroke and Vascular Neurology Medicine-Cardiology and Cardiovascular Medicine
CiteScore
11.20
自引率
1.70%
发文量
63
审稿时长
15 weeks
期刊介绍: Stroke and Vascular Neurology (SVN) is the official journal of the Chinese Stroke Association. Supported by a team of renowned Editors, and fully Open Access, the journal encourages debate on controversial techniques, issues on health policy and social medicine.
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