Administration of hypoxic pretreated adipose-derived mesenchymal stem cell exosomes promotes spinal cord repair after injury via delivery of circ-Astn1 and activation of autophagy

IF 4.7 2区医学 Q2 IMMUNOLOGY International immunopharmacology Pub Date : 2025-03-05 DOI:10.1016/j.intimp.2025.114324

Minghao Shao , Mingming Jin , Lv Feizhou , Xiaosheng Ma , Zhu Wei

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Abstract

Background

The aim of this study was to investigate the role and mechanism of exosomes isolated from adipose-derived mesenchymal stem cells (ADSCs) on spinal cord repair.

Methods

High-throughput sequencing was used to investigate abnormal expression of circular RNA (circRNA) in ADSC exosomes pretreated under hypoxic conditions (HExos) and ADSCs exosomes under normal conditions (Exos). The abnormal expression of mRNA in spinal cord tissues was also analyzed using high-throughput sequencing. Bioinformatics and luciferase reporter analyses were used to clarify the relationship among circRNA, micro RNA (miRNA), and mRNA. BV2 cells were used to analyze apoptosis levels and inflammatory cytokine expression under oxygen-glucose deprivation (OGD) conditions by using immunofluorescence and enzyme-linked immunosorbent assay (ELISAs). An SCI mouse model was also constructed and the therapeutic effect of Exos was detected using immunohistochemistry and immunofluorescence.

Results

High-throughput sequencing results showed that circ-Astn1 played a role in HExo-mediated spinal cord repair after SCI. Downregulation of circ-Astn1 decreased the therapeutic effect of HExos. We also found that Atg7 played a role in HExo-mediated spinal cord repair after SCI. Luciferase reporter analysis confirmed that both miR-138-5p and Atg7 were downstream targets of circ-Astn1. Downregulation of Atg7 or overexpression of miR-138-5p reversed the protective effect of circ-Astn1 on BV2 cells after exposure to OGD conditions. In contrast, upregulation of circ-Astn1 increased the therapeutic effects of Exo-mediated spinal cord repair after SCI via autophagy activation.

Conclusions

Taken together, the results indicate that ADSC-Exos containing circ-Astn1 promoted spinal cord repair after SCI by targeting the miR-138-5p/Atg7 pathway, which mediated autophagy.

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低氧预处理脂肪源性间充质干细胞外泌体通过递送circ-Astn1和激活自噬促进损伤后脊髓修复

本研究的目的是探讨从脂肪源性间充质干细胞（ADSCs）中分离的外泌体在脊髓修复中的作用和机制。方法采用高通量测序技术，研究缺氧预处理的ADSC外泌体（HExos）和正常处理的ADSC外泌体（Exos）中环状RNA （circRNA）的异常表达。采用高通量测序分析脊髓组织中mRNA的异常表达。利用生物信息学和荧光素酶报告基因分析来阐明circRNA、micro RNA （miRNA）和mRNA之间的关系。采用免疫荧光和酶联免疫吸附法（elisa）分析氧糖剥夺（OGD）条件下BV2细胞的凋亡水平和炎症细胞因子的表达。建立脊髓损伤小鼠模型，采用免疫组织化学和免疫荧光检测Exos的治疗效果。结果高通量测序结果显示circ-Astn1在hexo介导的脊髓损伤后修复中发挥作用。circ-Astn1的下调降低了HExos的治疗效果。我们还发现Atg7在脊髓损伤后hexo介导的脊髓修复中发挥作用。荧光素酶报告基因分析证实miR-138-5p和Atg7都是circ-Astn1的下游靶点。在暴露于OGD条件后，Atg7的下调或miR-138-5p的过表达逆转了circ-Astn1对BV2细胞的保护作用。相反，circ-Astn1的上调增加了exo介导的脊髓自噬激活后脊髓损伤修复的治疗效果。综上所述，结果表明含有circ-Astn1的ADSC-Exos通过靶向介导自噬的miR-138-5p/Atg7通路促进脊髓损伤后的脊髓修复。

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来源期刊

International immunopharmacology 医学-免疫学

CiteScore

8.40

自引率

3.60%

发文量

935

审稿时长

53 days

期刊介绍： International Immunopharmacology is the primary vehicle for the publication of original research papers pertinent to the overlapping areas of immunology, pharmacology, cytokine biology, immunotherapy, immunopathology and immunotoxicology. Review articles that encompass these subjects are also welcome. The subject material appropriate for submission includes: • Clinical studies employing immunotherapy of any type including the use of: bacterial and chemical agents; thymic hormones, interferon, lymphokines, etc., in transplantation and diseases such as cancer, immunodeficiency, chronic infection and allergic, inflammatory or autoimmune disorders. • Studies on the mechanisms of action of these agents for specific parameters of immune competence as well as the overall clinical state. • Pre-clinical animal studies and in vitro studies on mechanisms of action with immunopotentiators, immunomodulators, immunoadjuvants and other pharmacological agents active on cells participating in immune or allergic responses. • Pharmacological compounds, microbial products and toxicological agents that affect the lymphoid system, and their mechanisms of action. • Agents that activate genes or modify transcription and translation within the immune response. • Substances activated, generated, or released through immunologic or related pathways that are pharmacologically active. • Production, function and regulation of cytokines and their receptors. • Classical pharmacological studies on the effects of chemokines and bioactive factors released during immunological reactions.