Degenerated nucleus pulposus cells derived exosome carrying miR-27a-3p aggravates intervertebral disc degeneration by inducing M1 polarization of macrophages.
Degenerated nucleus pulposus cells derived exosome carrying miR-27a-3p aggravates intervertebral disc degeneration by inducing M1 polarization of macrophages.
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携带miR-27 a-3 p的退变髓核细胞来源的外泌体通过诱导巨噬细胞的M1极化来减轻椎间盘退变。
DOI：
10.1186/s12951-023-02075-y
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发表时间：
2023-09-04
影响因子：
10.2
通讯作者：
Ye, Zhengxu
中科院分区：
文献类型：
--
作者：
Zhao, Xin;Sun, Zhen;Xu, Benchi;Duan, Wei;Chang, Le;Lai, Kangwei;Ye, Zhengxu
椎间盘退变（IVDD）是脊柱疾病的主要原因。既往研究表明，免疫细胞，特别是巨噬细胞的浸润，在IVDD的进展中起着至关重要的作用。外泌体（exo）被认为在细胞间通讯中发挥着重要作用。本研究旨在探讨退变髓核（dNPc）来源的外泌体在巨噬细胞M1极化过程中的作用。从椎间盘退变（IVDD）和特发性脊柱侧凸患者中收集髓核（NP）组织和髓核细胞（NPc）。进行免疫组织化学分析以确定 NP 组织中 M1 巨噬细胞的数量。随后，收集来自退化NP细胞（dNPc-exo）和非退化NP细胞（nNPc-exo）的外泌体，并与从THP-1细胞诱导的M0巨噬细胞共培养。使用蛋白质印迹、流式细胞术、免疫荧光染色和 qRT-PCR 评估 M1 表型。通过RNA测序分析检测dNPc-exo和nNPc-exo组中microRNA的表达水平，并通过qRT-PCR研究不同microRNA诱导巨噬细胞极化的效果。此外，采用蛋白质印迹和qRT-PCR来证明dNPc-exo携带的microRNA对巨噬细胞下游靶信号通路的调节作用。最后，利用IVDD动物模型研究dNPc-exo对诱导巨噬细胞M1极化的影响及其在IVDD过程中的作用。在这项研究中，我们观察到随着椎间盘 (IVD) 退化，M1 巨噬细胞的数量增加。此外，我们发现dNPc释放的外泌体（dNPc-exo）可以促进巨噬细胞向M1表型极化。值得注意的是，通过对 dNPc-exo 和 nNPc-exo 组的 RNA 测序分析，我们发现 miR-27a-3p 是 dNPc-exo 组中高表达的 miRNA，它显着影响巨噬细胞 M1 极化的诱导。然后，我们发现dNPc-exo具有转运miR-27a-3p并靶向PPARγ/NFκB/PI3K/AKT信号通路的能力，从而影响巨噬细胞的M1极化。我们使用 IVDD 大鼠模型进行实验，观察到携带 miR-27a-3p 的外泌体实际上诱导了巨噬细胞的 M1 极化，并加剧了 IVD 的降解。总之，我们的研究结果强调了 dNPc-exo 在 IVDD 过程中的重要作用，并为进一步研究 IVDD 机制和基于外泌体的治疗潜力提供了基础。在线版本包含可在 10.1186/s12951-023-02075-y 获取的补充材料。
Intervertebral disc degeneration (IVDD) is a major contributor to spinal disorders. Previous studies have indicated that the infiltration of immunocytes, specifically macrophages, plays a crucial role in the advancement of IVDD. Exosomes (exo) are believed to play a significant role in intercellular communication. This study aims to investigate the role of exosomes derived from degenerated nucleus pulposus (dNPc) in the process of macrophages M1 polarization. Nucleus pulposus (NP) tissue and nucleus pulposus cells (NPc) were collected from patients with intervertebral disc degeneration (IVDD) and idiopathic scoliosis. Immunohistochemistry analysis was performed to determine the number of M1 macrophages in NP tissue. Subsequently, exosomes derived from degenerated NP cells (dNPc-exo) and non-degenerated NP cells (nNPc-exo) were collected and co-cultured with M0 macrophages, which were induced from THP-1 cells. The M1 phenotype was assessed using western blot, flow cytometry, immunofluorescence staining, and qRT-PCR. RNA-sequencing analysis was conducted to examine the expression levels of microRNAs in the dNPc-exo and nNPc-exo groups, and qRT-PCR was performed to investigate the effect pf different microRNA to induce macrophage polarization. Furthermore, western blot and qRT-PCR were employed to demonstrate the regulatory effect of microRNAs carried by dNPc-exo on downstream target signaling pathways in macrophages. Finally, an animal model of IVDD was utilized to investigate the impact of dNPc-exo on inducing M1 polarization of macrophages and its role in the IVDD process. In this study, we observed an increase in the number of M1 macrophages as the intervertebral disc (IVD) degraded. Additionally, we discovered that dNPc releases exosomes (dNPc-exo) could promote the polarization of macrophages towards the M1 phenotype. Notably, through RNA-sequencing analysis of dNPc-exo and nNPc-exo groups, we identified miR-27a-3p as a highly expressed miRNA in the dNPc-exo group, which significantly influences the induction of M1 polarization of macrophages. And then, we discovered that dNPc-exo has the ability to transport miR-27a-3p and target the PPARγ/NFκB/PI3K/AKT signaling pathway, thereby influencing the M1 polarization of macrophages. We conducted experiments using rat model of IVDD and observed that the exosomes carrying miR-27a-3p actually induced the M1 polarization of macrophages and exacerbated the degradation of IVD. In conclusion, our findings highlight the significant role of dNPc-exo in IVDD process and provide a basis for further investigation into the mechanism of IVDD and the potential of exosome-based therapy. The online version contains supplementary material available at 10.1186/s12951-023-02075-y.
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影响因子：
5
作者：
Feng X;Li Y;Su Q;Tan J
通讯作者：
Tan J
DOI：
10.3390/ph16060783
发表时间：
2023-05-24
期刊：
Pharmaceuticals (Basel, Switzerland)
影响因子：
--
作者：
Wu X;Sun L;Xu F
通讯作者：
Xu F
DOI：
10.1126/science.aau6977
发表时间：
2020-02-07
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Kalluri R;LeBleu VS
通讯作者：
LeBleu VS
影响因子：
6
作者：
Domoto R;Sekiguchi F;Tsubota M;Kawabata A
通讯作者：
Kawabata A
影响因子：
3.1
作者：
Capossela, S.;Schlafli, P.;Stoyanov, J. V.
通讯作者：
Stoyanov, J. V.