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间充质干细胞衍生和 siRNA 包裹的外泌体抑制股骨头坏死。

Mesenchymal stem cells-derived and siRNAs-encapsulated exosomes inhibit osteonecrosis of the femoral head.

机构信息

Department of Orthopaedics, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China.

出版信息

J Cell Mol Med. 2020 Sep;24(17):9605-9612. doi: 10.1111/jcmm.15395. Epub 2020 Aug 4.

DOI:10.1111/jcmm.15395
PMID:32749049
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7520260/
Abstract

Osteonecrosis of the femoral head (ONFH) is a progressive, obstinate and disabling disease. At present, the treatment of ONFH is still a global medical problem. We aim to explore the role of bone mesenchymal stem cells (BMSCs)-derived and siRNAs-encapsulated exosomes (siRNAs-encapsulated BMSCexos) in ONFH. We first isolated BMSCexos and screened siRNAs of 6 ONFH-related genes for siRNAs-encapsulated BMSCexo. The expression of these 6 ONFH-related genes in dexamethasone (DXM)-treated MC3T3-E1 cell, cell model of ONFH, was detected by RT-qPCR and Western blot analysis. And then, we performed CCK-8 assay, angiogenesis assay and HE staining analysis to test the promotion role of the siRNAs-encapsulated BMSCexo for angiogenesis during ONFH repair. The results suggest that the obtained particles were BMSCexos. The screened effective siRNAs could effectively knock down their expression in VECs. Moreover, siRNAs-encapsulated BMSCexo could effectively knock down the expression of these genes in VECs. In addition, siRNAs-encapsulated BMSCexo promote angiogenesis during ONFH repair. In conclusion, we found siRNAs-encapsulated BMSCexos could promote ONFH repair by angiogenesis, and indicated exosome as the new siRNA carrier is of great significance to improve the efficiency of RNAi.

摘要

股骨头坏死（ONFH）是一种进行性、顽固性和致残性疾病。目前，ONFH 的治疗仍然是一个全球性的医学难题。我们旨在探讨骨间充质干细胞（BMSCs）衍生的和 siRNAs 包裹的外泌体（siRNAs 包裹的 BMSCexos）在 ONFH 中的作用。我们首先分离了 BMSCexos，并筛选了 6 个 ONFH 相关基因的 siRNAs 用于 siRNAs 包裹的 BMSCexo。通过 RT-qPCR 和 Western blot 分析检测了这些 6 个 ONFH 相关基因在地塞米松（DXM）处理的 MC3T3-E1 细胞（ONFH 细胞模型）中的表达。然后，我们进行了 CCK-8 测定、血管生成测定和 HE 染色分析，以测试 siRNAs 包裹的 BMSCexo 在 ONFH 修复过程中促进血管生成的作用。结果表明，获得的颗粒是 BMSCexos。筛选出的有效 siRNAs 可以有效地在 VECs 中敲低它们的表达。此外，siRNAs 包裹的 BMSCexo 可以有效地在 VECs 中敲低这些基因的表达。此外，siRNAs 包裹的 BMSCexo 促进 ONFH 修复过程中的血管生成。总之，我们发现 siRNAs 包裹的 BMSCexo 可以通过血管生成促进 ONFH 的修复，并表明外泌体作为新的 siRNA 载体对提高 RNAi 的效率具有重要意义。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e74e/7520260/17db4cbe75eb/JCMM-24-9605-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e74e/7520260/3eb149a4bdab/JCMM-24-9605-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e74e/7520260/b3fd1dd0565f/JCMM-24-9605-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e74e/7520260/d15fa94dd94a/JCMM-24-9605-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e74e/7520260/17db4cbe75eb/JCMM-24-9605-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e74e/7520260/3eb149a4bdab/JCMM-24-9605-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e74e/7520260/b3fd1dd0565f/JCMM-24-9605-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e74e/7520260/d15fa94dd94a/JCMM-24-9605-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e74e/7520260/17db4cbe75eb/JCMM-24-9605-g004.jpg

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