外泌体 miRNA-499a-5p 靶向 CD38 减轻蒽醌诱导的心脏毒性：实验研究。

Exosomes miRNA-499a-5p targeted CD38 to alleviate anthraquinone induced cardiotoxicity: experimental research.

机构信息

State Key Laboratory of Trauma, Burns and Combined Injury, Shock and Transfusion Research Department of Army Medical Center, Army Medical University, Chongqing.

Department of Cardiothoracic Surgery, Children's Hospital of Nanjing Medical University, Nanjing.

出版信息

Int J Surg. 2024 Apr 1;110(4):1992-2006. doi: 10.1097/JS9.0000000000001118.

DOI:10.1097/JS9.0000000000001118

PMID:38277348

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11019978/

Abstract

BACKGROUND

The purpose of this study was to investigate the effects of cardiac homing peptide (CHP) engineered bone marrow mesenchymal stem cells (BMMSc) derived exosomes (B-exo) loaded miRNA-499a-5p on doxorubicin (DOX) induced cardiotoxicity.

METHODS

miRNA chip analysis was used to analyze the differences between DOX induced H9c2 cells and control group. CHP engineering was performed on BMMSc derived exosomes to obtain C-B-exo. miRNA-499a-5p mimic was introduced into C-B-exo by electroporation technology to obtain C-B-exo-miRNA-499a-5p. DOX was used to establish a model of cardiotoxicity to evaluate the effects of C-B-exo- miRNA-499a-5p in vivo and in vitro . Western blot, immunohistochemistry, immunofluorescence, and other molecular biology methods were used to evaluate the role and mechanism of C-B-exo-miRNA-499a-5p on DOX induced cardiotoxicity.

RESULTS

miRNA chip analysis revealed that miRNA-499a-5p was one of the most differentially expressed miRNAs and significantly decreased in DOX induced H9c2 cells as compared to the control group. Exo-and B-exo have a double-layer membrane structure in the shape of a saucer. After engineering the CHP of B-exo, the results showed that the delivery of miRNA-499a-5p significantly increased and significantly reached the target organ (heart). The experimental results showed that C-B-exo-miRNA-499a-5p significantly improved electrocardiogram, decreased myocardial enzyme, serum and cardiac cytokines, improved cardiac pathological changes, inhibited CD38/MAPK/NF-κB signal pathway.

CONCLUSIONS

In this study, C-B-exo-miRNA-499a-5p significantly improved DOX-induced cardiotoxicity via CD38/MAPK/NF-κB signal pathway, providing a new idea and method for the treatment of DOX induced cardiotoxicity.

摘要

背景

本研究旨在探讨心脏归巢肽（CHP）工程骨髓间充质干细胞（BMMSc）衍生的外泌体（B-exo）负载 miRNA-499a-5p 对阿霉素（DOX）诱导的心脏毒性的影响。

方法

采用 miRNA 芯片分析 DOX 诱导的 H9c2 细胞与对照组之间的差异。对 BMMSc 衍生的外泌体进行 CHP 工程化，得到 C-B-exo。通过电穿孔技术将 miRNA-499a-5p 模拟物引入 C-B-exo，得到 C-B-exo-miRNA-499a-5p。用 DOX 建立心脏毒性模型，评估 C-B-exo-miRNA-499a-5p 在体内和体外的作用。采用 Western blot、免疫组织化学、免疫荧光等分子生物学方法评价 C-B-exo-miRNA-499a-5p 对 DOX 诱导的心脏毒性的作用及机制。

结果

miRNA 芯片分析显示，miRNA-499a-5p 是差异表达最显著的 miRNA 之一，与对照组相比，在 DOX 诱导的 H9c2 细胞中显著下调。外泌体和 B-exo 呈碟形双层膜结构。对 B-exo 的 CHP 工程化后，结果表明 miRNA-499a-5p 的递送明显增加，并明显到达靶器官（心脏）。实验结果表明，C-B-exo-miRNA-499a-5p 明显改善心电图，降低心肌酶、血清和心脏细胞因子，改善心脏病理变化，抑制 CD38/MAPK/NF-κB 信号通路。