青春期抑郁样行为小鼠脑组织外泌体miRNA测序分析

曾琪; 廖婧; 冉柳毅; 石磊; 陈羽佳; 张晨钰; 向娇娇; 洪素; 况利

doi:10.12182/20230360205

青春期抑郁样行为小鼠脑组织外泌体miRNA测序分析

Sequencing Analysis of miRNAs in Brain-Derived Exosomes of Adolescent Mice With Depression-Like Behaviors

摘要

摘要:
目的探索青春期抑郁样行为小鼠脑组织外泌体微RNA（microRNA, miRNA）差异表达情况。
方法实验组为慢性社交挫败实验应激模型（chronic social defeat stress, CSDS）敏感型青春期小鼠，采用糖水偏好和旷场实验评估抑郁样行为。使用超速离心法提取脑组织外泌体，经透射电子显微镜、纳米流式检测技术以及蛋白质印记对外泌体形态、粒径大小和表面标志蛋白进行鉴定。采用高通量测序技术评估实验组和对照组小鼠脑组织外泌体miRNAs表达，基于生物信息学进行GO和KEGG通路富集分析。
结果外泌体颗粒大小在50～100 nm之间、呈典型圆盘状的囊泡结构，检测到外泌体阳性蛋白TSG101和Syntenin。CSDS诱导抑郁样行为的青春期小鼠脑组织外泌体中有13个miRNA显著上调，4个miRNA显著下调，差异表达的miRNA在PI3K-Akt信号通路、轴突导向以及缺氧反应等显著富集。
结论本研究发现脑组织外泌体miRNA可能参与胰岛素抵抗、神经可塑性、缺氧反应等生物过程调控大脑功能，从而产生抑郁样行为。

Abstract:
Objective To explore the differential expression of microRNAs (miRNAs) in brain-derived exosomes (BDEs) of adolescent mice with depression-like behavior.
Methods The experimental group consisted of susceptible adolescent mice exposed to chronic social defeat stress (CSDS), and sucrose preference test (SPT) and open field test (OFT) were performed to evaluate their depression-like behaviors. BDEs were extracted by ultracentrifugation (UC). The morphology, particle size, and surface marker proteins of BDEs were examined by transmission electron microscopy, nano-flow cytometry and Western blot. The expression of miRNA in BDEs was evaluated by high-throughput RNA sequencing. GO enrichment analysis and KEGG pathway enrichment analysis were carried out based on bioinformatics.
Results The particle size of BDEs ranged between 50 to 100 nm and they displayed a typical disc-shaped vesicle structure. TSG101 and syntenin, the exosome-positive proteins, were detected. In the BDEs of mice with depression-like behaviors induced by CSDS, 13 miRNAs were significantly upregulated and 4 miRNAs were significantly downregulated. Go and KEGG analysis showed that differentially expressed miRNAs were significantly enriched in PI3K-Akt signaling pathway, axonal guidance, and hypoxic response.
Conclusion It was found in this study that exosomal miRNAs in brain tissue might be involved in such biological processes as insulin resistance, neuroplasticity, and hypoxic response, thereby regulating brain functions and causing depression-like behaviors.

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