诱导多能干细胞分泌的外泌体通过长非编码 RNA-0949 减轻脂多糖诱导的神经炎症反应。

Exosomes secreted from induced pluripotent stem cell ameliorate the lipopolysaccharide induced neuroinflammatory response via lncRNA-0949.

机构信息

Department of Respiratory and Critical Care Medicine, the First Affiliated Hospital of Nanchang University, Nanchang, Jian, People's Republic of China.

Department of Respiratory and Critical Care Medicine, Nanchang, Jiangxi Province, People's Republic of China.

出版信息

Immun Inflamm Dis. 2024 Mar;12(3):e1155. doi: 10.1002/iid3.1155.

DOI:10.1002/iid3.1155

PMID:38533916

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10966914/

Abstract

PURPOSE

To study the effect of exosomes derived from the induced pluripotent stem cells (iPSCs) in the neuroinflammatory response of microglia caused by lipopolysaccharide (LPS) and reveal the potential underlying mechanism.

METHODS

A permanent microglia cell line HMO6 was activated by LPS. The features of exosomes were analyzed by nano flow cytometry, Western blot and transmission electron microscope. The RNA-seq was used to analyze the difference of noncoding RNA profiles between iPSC-Exos and HMO6 derived exosomes and proved that long no-coding RNA (lncRNA-0949) was highly expressed in the iPSC-Exos. Activated HMO6 cells were cocultured with iPSC-Exos in which lncRNA-0949 was overexpressed, knocked down or normally expressed. Quantitative real-time polymerase chain reaction (RT-qPCR), Enzyme-Linked Immunosorbent Assay and Western blot assay were adopted to analyze RNA and protein expression of inflammatory factors in HMO6 cells.

RESULTS

The oxidative stress and inflammatory response of microglia were significantly attenuated with the iPSC derived exosomes treatment. LncRNA-0949 was effectively delivered into the HMO6 cells through the iPSC-Exos, which largely alleviated the production of malondialdehyde, IL-6, IL-1β and TNF-α in HMO6 cells. Overexpression of lncRNA-0949 could enhance the anti-inflammatory effect of the iPSC-Exos, and knock-down of lncRNA-0949 impaired this availability.

CONCLUSION

According to our results, lncRNA-0949 enriched exosomes from iPSC could potentially be used as a therapeutic strategy to prevent/treat neuroinflammatory diseases.

摘要

目的

研究诱导多能干细胞（iPSC）衍生的外泌体对脂多糖（LPS）引起的小胶质细胞神经炎症反应的影响，并揭示其潜在的作用机制。

方法

用 LPS 激活永生化小胶质细胞系 HMO6。采用纳米流式细胞术、Western blot 和透射电镜分析外泌体的特征。通过 RNA-seq 分析 iPSC-Exos 和 HMO6 衍生的外泌体之间非编码 RNA 谱的差异，并证明长非编码 RNA（lncRNA-0949）在 iPSC-Exos 中高度表达。用过表达、敲低或正常表达 lncRNA-0949 的 iPSC-Exos 与激活的 HMO6 细胞共培养。采用实时定量聚合酶链反应（RT-qPCR）、酶联免疫吸附试验和 Western blot 检测分析 HMO6 细胞中炎症因子的 RNA 和蛋白表达。

结果

iPSC 衍生的外泌体处理可显著减轻小胶质细胞的氧化应激和炎症反应。lncRNA-0949 通过 iPSC-Exos 有效地转染到 HMO6 细胞中，从而显著减少 HMO6 细胞中丙二醛、IL-6、IL-1β和 TNF-α的产生。lncRNA-0949 的过表达可增强 iPSC-Exos 的抗炎作用，而 lncRNA-0949 的敲低则削弱了这种作用。